-->Plant Physiology and Molecular Biology

Effects of Cr⁶⁺on the growth and some physiological characters of tobacco by tissue culture

SHI Gui-Yu, QIN Li-Feng, CHEN Geng-Yun — 2016/1/15 21:08:50

After germfree tobacco seedlings being cultivated in culture medium added with Cr⁶⁺ solution in 0,50,100,150,200,300 μmol·L^-1 consistency respectively for 35 days,the growth and certain physiological functions of the tobacco were studied. The results indicated:(1)the callus and seedlings growth was promoted at 50 μmol·L^-1 consistency,and the height,fresh weight,content of chlorophyll and protein, SOD(superoxide dismutase)activity of the seedlings increased;(2)the growth of callus and seedling was restrained distinctly under the high consistency(100～300 μmol·L^-1),and the seedling height,fresh weight,content of chlorophyll and protein,activity of SOD(superoxide dismutase)decreased;(3)on the contrary,the activity of peroxidase(POD),content of MDA and the permeability of cell membrane had earned a remarkable increase with the high concentration of Cr⁶⁺.

A genetic transformation of roselle callus mediated by Agrobacterium tumefaciens

2016/1/15 21:08:50

In present paper,the basic conditions for the genetic transformation of Roselle callus were investigated by using Agrobacterium tumefaciens LBA4404 and EHA105,harboring pBI121,as donor strains and a genetic transformation system of Roselle callus was initiatively established,by which two transgenic Roselle cell lines expressed stably NPTⅡ activity were obtained. The results of GUS histochemical assays and PCR amplification demonstrated that the overall transformation rate of Roselle callus was 4%. It is feasible to use Agrobacterium-mediated method to introduce exogenous gene into Roselle cells via callus

Study on asymbiotic germination of Cattleya hybrid seed and development of protocorms during germinating

DING Lan, WANG Li, LI Huai, QI Lin-Lin, LI Juan — 2016/1/15 21:08:50

This paper presented the optimum culture conditions for the asymbiotic germination of the seeds of Cattleya hybrid and the development process of protocorms during germinating was also investigated. Mediums with lower ionic concentrations, such as KC and RE, were suitable for seed germination. KC medium supplemented with 1.0 mg/L BA and 0.1 mg/L NAA or 1.0 mg/L KT and 0.1 mg/L NAA was the optimal choice. Cytological observation indicated that the embryonic cells of the seeds developed into protocorms through cell division, then differentiated into leaf primordium, buds and finally integral seedlings.

Technology of accelerating germination and desiccation tolerance in Archontophoenix alexandrae seeds

2016/1/15 21:08:51

In order to improve the artificial planting technology of Archontophoenix alexandrae,different chemical treatments to accelerate seed germination were carried out to seek efficient methods of accelerating germination, the seed desiccation tolerance were also investigated in this study. The results showed that soaking in solutions of 20% H₂O₂ and 98% H₂SO₄ for 5 min,0.3% NaNO₂ and KNO₃ for 24h all could improve germination percentage and velocity,especially in 98% H₂SO₄ and 0.3% NaNO₂ solution. Soaking in 200～1 000 mg/L GA and 20～100 mg/L KT solution for 24 h could also accelerate germination,but the efficiency of accelerating was related to the concentration of solutions. The germination percentage of mature seeds increaced slightly after slight desiccation,but decreased significantly when the seed moisture content reduced to below 17% and then fully lost when below 10%,so the seeds tend to be intermediate.

Characteristics of chlorophyll fluorescence and comparison of drought resistance among different species of karst plants

2016/1/15 21:08:51

Under different actinic light intensities,the characteristics of light energy using were observed in five species of karst plants by using technique of chlorophyll fluorescence,and their drought resistance was compared under water stress induced by PEG. The five karst plants are Cladrastis platycarpa,Impatiens macrovexilla,Alchornea trewioides,Chirita eburnea and Quercus glauca. The results showed that qP decreased gradually and NPQ increased evidently in these plants with the increase of actinic light intensity. Simultaneously,Fv'/Fm' and ΦPSⅡ declined significantly. However,there was no evident difference in fluorescence characteristics between these plants under the same actinic light. The Fv/Fm of Chirita eburnea and Quercus glauca changed insignificantly under the water stress induced by PEG, and the Fv/Fm decreased moderately in Cladrastis platycarpa and Impatiens macrovexilla,but the value decreased most evidently in Alchornea trewioides. Thus,it was deduced that Chirita eburnea and Quercus glauca are the strongest in drought resistance,Alchornea trewioides is the lowest,Cladrastis platycarpa and Impatiens macrovexilla are in the middle.

Seasonal photosynthetic responses of Phyllostachys edulis to environmental factors

2016/1/15 21:08:51

By using Li-6400,the seasonal photosynthetic response of Phyllostachys edulis to light intensity,CO₂ concentration,temperature and humidity were determined. The main results are as follows:the annual average of maximal net photosynthetic rate(P_max),light compensate point(LCP),light saturate point(LSP),and apparent quantum yield(AQY)for P.edulis are respectively 7.30,19.15,1 075 mmol·m^-2·s^-1 and 0.032. The seasonal photosynthetic variation tendency of P.edulis to light intensity were summer>autumn>winter>spring for P_max,spring>summer>winter>autumn for LCP,and autumn>summer>winter>spring for LSP and AQY. The annual average of CO₂ compensate point(CCP),CO₂ saturate point(CSP)and carboxylase efficiency(CE)for P.edulis are respectively 73.52,1 500 μmol·mol^-1 and 0.033. The seasonal photosynthetic variation tendency of P.edulis to CO₂ concentration were spring>winter>autumn>summer for LCP,spring>autumn>summer>winter for CCP,and summer>autumn>winter>spring for CE. The optimal temperature for photosynthesis of P.edulis ranges from 20 to 30 ℃,equivalent in spring and autumn,higher in summer and lowest in winter. It is close to the average of three day's highest temperature before test in spring and autumn,and close to the average of ten day's highest temperature before test in summer and winter. The optimal humidity for photosynthesis of P.edulis ranges from 40% to 65%,its seasonal variation tendency is autumn>summer>winter>spring. On the whole,the seasonal photosynthetic responses of P.edulisto environmental factors were closely related with seasonal change of the environment and the physiological activity of leaves.

Comparison of caloric values and ash contents in three palm species

2016/1/15 21:08:51

An experiment was conducted to evaluate the caloric value and ash contents in various components of three palm species(Butia capitata,Elaeis guineensis,Acoelorraphe wrightii)from the nursery in the Xiamen Botanical Garden. The results showed that the gross caloric values(GCV)of Butia capitata in various components can be ranked in the decreasing order as follows:mature leaves>lower roots>young leaves>upper roots>rhizome>petioles; for Acoelorraphe wrightii,the order was:mature leaves>upper roots>lower roots>young leaves>petioles; for E.guineensis,the order was:mature leaves>upper roots>lower roots>young leaves>petioles>rhizomes. The ash free caloric values(AFCV)in different components also varied widely,and could be ranked almost in the same order as those of GCV. B.capitata ash contents in various components could be ranked in a decreasing order as follows:lower roots>upper roots>rhizomes>young leaves>petioles>mature leaves; for A.wrightii,the order was:young leaves>mature leaves>lower roots>upper roots>petioles; for E.guineensis,the order was:mature leaves>young leaves>lower roots>upper roots>petioles>rhizomes. The results indicated that the cold resistant B.capitata has a more reasonable distribution of caloric values and ash contents,this character may help to improve its adaptation to the low temperature.

Effects of light on maca callus growth, shoot induction and its survival rate

2016/1/15 21:08:51

Maca(Lepidium meyenii),belongs to the family of Brassicaceae,has high value of nutrition and medicine. In the process of micropropagation,light had remarkable influences on maca callus growth,shoot induction and survival. Green light and blue light not only made against maca callus growth but also inhibited shoot induction and survival. White light,red light and yellow light improved callus growth remarkably. Under these light conditions,the shoot induction rates were from 60% to 80%,and the shoot survival rates were from 29% to 36%.Appropriate prolonging of the time under light increased the shoot induction rate,and the appropriate photoperiod was 16h per day. But higher intensity of light decreased the survival rate of the shoots,and the appropriate light intensity was 24-41 μmol/m²·s.

Rapid propagation of Dracaena cambodiana in vitro

2016/1/15 21:08:51

In this study,the three-month old Dracaena cambodiana about 25 cm high was used as the basic material. The tender stem fragments were cleaved lengthways into four pieces,each piece was used as the plantlet and induced in the medium modified MS supplemented with 6-BA2.0 mg/L and NAA0.1 mg/L,the germination rate is 78% after a month culture. The proliferation multiple of the bud is 2.9 after the germination was continuously induced twice in the same medium MS supplemented with 6-BA1.5 mg/L and NAA0.2 mg/L after two months,the medium was renovated once a month. What's more,the effect that the different form of bud used as a plantlet for subculture had on the proliferation multiple of the burgeon of Dracaena cambodiana when they were induced on the same medium has also been proved in this paper. Our result showed that the proliferation multiple of the clustering buds was 4.6 and single bud 0.8 when they were cultured on the same medium MS supplemented with 6-BA1.0 mg/L and NAA0.2 mg/L for a month. The rooting rate was 100% when the germination were cultured on the modified medium MS supplemented with NAA0.2 mg/L. And the survival rate of outplanting in nursery is 100%.

Shoot regeneration from cotyledon of orange-heading Chinese Cabbage

2016/1/15 21:08:35

The effects of hormone combination and AgNO₃ on adventitious shoot regeneration from cotyledon of orange-heading Chinese Cabbage were studied. The results showed that the adventitious shoot could not be induced from the cotyledon in MS medium added with cell division hormone(6-BA or TDZ)only; the regeneration frequency of shoot reached to 15% in MS medium added with cell division hormone(6-BA or TDZ)and growth hormone(NAA)together; the regeneration frequency could increase 42.5% in MS medium added with cell division hormone(TDZ),growth hormone(NAA)and AgNO₃. Compared with 6-BA,TDZ had more effective in inducing shoot regeneration. The maximum frequency of shoot regeneration(50%)was obtained in MS medium which contained 0.05 mg/L TDZ,0.3 mg/L NAA and 8 mg/L AgNO₃.

Organic nutrition and the shoot growing trait of heavily retractive pruned longan trees

CHEN Deng-Ke, LU Mei-Ying, TANG Teng, YU Huang-Sen — 2016/1/15 21:08:35

The shoot length,chlorophyll content,net photosynthetic rate,total soluble sugar content and the soluble protein content of each shoots of the heavily retractive pruned longan trees were determined,the result showed that the chlorophyll content and the soluble protein content between heavily retractive pruned longan trees and control had no significant difference,then the total soluble sugar content of the heavily retractive pruned longan trees was significantly less than that of the control,but the net photosynthetic rate was totally higher than that of the control while the shoots were significantly longer. PP₃₃₃applying can availably restrain watershoot and has no significant effect on other items which have been determined.

Cryopreservation of recalcitrant seed germplasm

2016/1/15 21:08:35

Being sensitive to desiccation and low temperature,the recalcitrant seeds can not be stored with conventional methods. However,cryopreservation has been considered as the most effective means. As far as cryostorage of recalcitrant seeds is concerned,vitrification and two-step freezing have been applied in many studies. Unfortunately,there are some factors,including water content of seeds or embryonic axes,effects of cryopretectants in solutions,cooling rate and thawing,rehydration and after-thawing culture that influence succeeding in cryopreservation. To produce normal plants after cryopreservation of recalcitrant seeds or embryonic axes,a carefully defined sequence of manipulations is needed to be solved. Besides the mechanisms of desiccation tolerance and sensitivity to low temperatures,the freezing and antifreezing mechanisms of plant cells are also ascertained imminently in order to dig out the most effective long-term storage method through seed-gene bank.

Metabolomics approaches and their application in Botany

DONG Deng-Feng — 2016/1/15 21:08:35

Metabolites are the end products of biological systems response to genetic control and/or environmental changes. Metabolomics,which study of total metabolites(metabolome),is an inevitable‘omics'following by genomics and proteomics. In this paper,metabolomics approaches including metabolites qualitative and quantitative determination,data acquisition and mining as well as their application in Botany such as target analysis,gene function determination,metabolite pathways and metabolic engineering,integrity Botany,signal transduction etc were reviewed.

Comparative analysis on effect of Fe²⁺,Cu²⁺and Zn²⁺ to physiological characteristics of plant

2016/1/15 21:08:35

This paper studied on seven physiological indices,which had correlation with stress effect of Fe²⁺,Cu²⁺and Zn²⁺ to plant when the treatment of soaking in water as control. Meanwhile,the principle and method of biochemistry and biostatistics were applied. The effect of three ions treatments to plant physiological characteristics was analyzed according to damage of oxygen free radicals. The results were as follows:the effect of physiological characteristics had evidently difference because different ion treatments had different stress damage. And the difference had marked correlation with ion stress character treated.

Study on changes in concentrations of four endogenous hormones in different fruit positions during citrus fruit growth and development

2016/1/15 21:08:35

Changes in concentrations of indol-3-acetic acid(IAA),abscisic acid(ABA),gibberellin(GA_1/3)and ribosylzeatin(ZR)were investigated in different fruit positions(rind,pulp and seed)during fruit growth and development of parthenocarpic‘Guoqing 1'satsuma mandarin and self-pollinated‘Huanong Bendizao'tangerine. The results were showed that concentrations of IAA,GA_1/3 and ZR in the rind of‘Guoqing 1'were relatively high during young fruit growth,and then IAA in the rind and pulp was decreased progressively,whereas significant peaks were observed for ABA in the pulp and GA_1/3,ZR in the rind and pulp during fruit enlargement,and ABA concentrations were obviously increased in the rind and pulp during fruit maturation. In‘Huanong Bendizao',notable peaks were found for IAA in the seed,rind and pulp and seed ABA at the stage of fruit enlargement,GA_1/3 and ZR concentrations in the seed were relatively high and significantly increased during fruitlet growth and early fruit enlargement,whereas the levels of four endogenous hormones in the rind and pulp were relatively low and changed a little as time passed. Relationship between fruit growth and development and dynamics of endogenous IAA,ABA,GA_1/3 and ZR concentrations in different fruit parts was discussed herein.

Rule of growth and development on flower-bud of Mangifera indica cv.Zihuamang

2016/1/15 21:08:35

The growth and development of mango(cv.Zihuamang)flower-bud were studied by using methods of girdinge,picking off leaves and paraffin section. The results showed that the physiological differentiation started after 1 to 6 weeks,in early November,when last mature shoots stop growing. More than 75% of buds passed the physiological differentiation stage until November 23th. Morphological differentiation stage began in middle November after the last mature shoots stop growing,until middle or late January in next year,and the morphological differentiation stage lasted for 60～75 days. Vegetative stage or the period of leaf bud was from October 30th to November 6th. Preceding differentiation of flower-bud was from November 16th to January 4th in next year. Inflorescence differentiation stage was from November 16th to January 8th in next year. The first branching of inflorescence was from December 21th to January 8th in next year. The second branching of inflorescence was from December 21th to January 14th in next year. The small flower organ of basal inflorescence began to differentiated from January 8th to January 26th in the next year. At first,the calyx and petal begin to differentiate,and then the stamen,pistil and honey plate. It's the differentiated period of flower organ.

Studies on the development of the leaf of Pinus kwangtungensis

2016/1/15 21:08:35

Optical microscope and electron microscope were applied in the study of the growth and development of the leaves of Pinus kwangtungensis. The results are as follows:the triangulate needle with simple venation,single-cyclocytic type of stomata on venterof the epidermis,one or two layers hypodermis,transfusion tissue of pine type,endodermis are obvious; three resin ducts formed five types:only two middle type,only two edge type,one middle or not type,three edge type,and three middle type; the serrulate margin of leaves developed from the epidermis'cell wall. The stomatal apparatus developed from the middle to the edges with mergence and discontinuance in some region.

Sex identification of Siraidia grosvenorii by isozyme analysis

2016/1/15 21:08:35

The peroxidase,esterase,superoxide dismutase,polyphenol oxidase and catalase isozyme of Siraidia grosvenorii were analyzed with vertical slab PAGE and specific cytochemical staining. The results showed that isozyme bands have apparent difference between the male and female plant. The number of different band of sex character was above one in each isozyme. It is concluded that the difference on isozyme bands can be used for sex identification of Siraidia grosvenorii. In addition,the isozyme bands and enzyme activity of high yield female plant,low yield female plant and acarpous female plant were compared.

Changes on leaf oxalate content in buckwheat growing under different nitrogen forms

2016/1/15 21:08:18

Buckwheat(Fagopyrum esculentum)seedling were pretreated for 3 d with 1/5 Hoagland culture solution,before the buckwheat seedling cultured to expanded primary leaves under hydroponic condition with ammonia nitrogen or nitrate nitrogen as the only nitrogen source,then oxalate content was determined in leaves,roots and root exudates. The results showed the oxalate content was very significantly decreased in buckwheat leaves when ammonia was used as the sole nitrogen source,so was in the root and its exudating rate. This implies that the oxalate decrease by ammonia results from the metabolic change rather than the change in transport to the root and in the exudating rate. The leaves had much less some organic acids,activities of several enzymes involved in oxalate metabolism when the plant grew under ammonia nitrogen,indicating these organic acids level involved in oxalate metabolism may be one of the factors controlling oxalate content. Based on the present data,it is concluded that oxalate accumulation in buckwheat leaves is closely related to some organic acids metabolism.

Establishment of genetic transformation system of Populus euphratica and optimization of antibiotic concentration

LI Wei, CHEN Xiao-Yang, DING Xia, LI Hui — 2016/1/15 21:08:18

Effects of hormones including NAA,IAA,6-BA on leaf differentiation of Populus euphratica and antibiotics including Kanamycin(Kan),G418,Carbenicillin(Cab),Cefotaxime(Cef)on the growth and differentiation or rooting of various explants of P.euphratica were studied. Genetic transformation system mediated by Agrobacterium and optimum antibiotic concentrations for modified organism selection of P.euphratica were founded. Results shown that:the optimal culture medium for leaf differentiation of P.euphratica was MS+BA 0.5 mg/L+NAA 0.1～0.2 mg/L+sugar 25 g/L+agar5 g/L; at co-culture stage which leaves were used as explants,the optimum concentrations of Kan,G418,Cab and Cef were 10 mg/L,7.5 mg/L,200～600 mg/L,200～400 mg/L respectively; at the stage of subculture or rooting of resistance buds,the optimum concentrations were 15～20 mg/L,10～15 mg/L,200～800 mg/L,200～600 mg/L respectively.

Inducible formation of 2-(2-phenylethyl)chromones in cell suspension culture of Aquilaria sinensis

2016/1/15 21:08:18

Aquilaria sinensis is the only species of Aquilaria in China for agarwood resources and has been recorded as endangered plant since 1992. The present dissertation was undertaken to study the inducible formation of the second metabolites in A.sinensis. Four 2-(2-phenylethyl)chromones existed in agarwood were identified. For the first time, fungal extracts of Menanotus flavolivens elicited the production of these chromones in cell suspension culture of A.sinensis,whereas they were not existing in normal condition. And a cell suspension culture system of A.sinensis was established.

Preliminary study on the effect of low pH on the nodulation of Medicago sativa and corresponded mechanisms

2016/1/15 21:08:18

Effects of low pH on root hair deformation(RHD)and nodulation in Medicago sativa were investigated and corresponded mechanisms was also discussed in present study. It was showed that primary root elongation,the OD₆₀₀ of Rhizobium meliloti,and symbiotic nodulation of roots were obviously decreased at low pH. At pH5.0,pH4.7,pH4.5,pH4.2,the rate of RHD was less 44.1%,56.4%,60.0% and 69.0% respectively than that at pH6.5 in the presence of R.meliloti. Moreover,in the presence of NF RHD was also less 45.9% at pH4.5 than that at pH6.5. These results implied that the nodulation signal transduction of Medicago sativa at initial stages of symbiotic nodulation was inhibited at low pH,which might be an important cause resulted in the reduction of nodulation in Medicago sativa at acidity conditions.

Relation between endogenous hormones in bud and on-year or off-year fruiting of Ginkgo biloba

2016/1/15 21:08:18

To find the physical causes of on and off-year fruiting of Ginkgo biloba,four kinds of endogenous hormones including IAA,ZR,GA and ABA in fruit bud and leaf bud of on-year and off-year fruiting trees were determined with ELISA from 2004 to 2005.The results are as follows:the content ratios of IAA,ZR and ABA in floral bud to leaf bud of on-year Ginkgo trees are greater than that of off-year trees,but the content ratio of GA in fruit bud to leaf bud of on-year trees is less than that of off-year trees,this will benefit the differentiation and growth of fruit bud,and promote on-year fruiting; high contents of ZR,IAA and ABA,and low content of GA,and high ratios of IAA/GA,ZR/GA and ABA/GA in fruit bud are helpful to fruit bud's differentiation and on-year fruiting; high contents of ZR and IAA,and low content of ABA,especially the increase of ZR in early or middle period of leaf bud are advantageous to on-year fruiting.

Effects of calcium signal system on anti-oxidant enzymes and membrane lipid peroxidation of citrus under low temperature

2016/1/15 21:08:18

Effects of calcium signal system on anti-oxidant enzymes and membrane lipid peroxidaton of citrus callus under low temperature were studied. The results showed that exogenous Ca²⁺(5 mmol/L)could markedly decrease membrane injury rate and malondialdehyde(MDA)accumulation during low temperature stress. Moreover, it could raise the content of soluble protein and increase the activity of superoxide dismutase(SOD)and peroxidase(POD).Exogenous calcium could strengthen the cold-resistant of citrus,compared with Ca-deficient treatment. But such effects were inhibited by addition of Ca²⁺ chelating agent EGTA or calmodulin antagonist TFP in the culture medium. Taken together the results above,it is suggested that calcium signal system may be involved in the membrane lipid peroxidation and protective enzyme system of cold-resistant of citrus under low temperature.

Effects of ethephon sprayed at early tillering stage on the activities of peroxidase, IAA oxidase and acid invertase in sugarcane in correlation to tillering

2016/1/15 21:08:18

Foliar application of 100 mg/L ethephon at tillering stage increased the activities of IAA oxidase and peroxidase in sugarcane plant. It was obvious that the activities of IAA oxidase and peroxidase in top internodes were higher as compared with those in bottom internodes. The treatment with 100 mg/L ethephon was the most efficacious in all the treatments. For ROC16, after ethephon treatment,the acid invertase activity in the top internodes was higher than that in the bottom internodes from beginning to end, and the treatment with 100 mg/L ethephon recorded higher activity in the bottom internodes than that in the control. For ROC22, both the treatments with 100 mg/L and 400 mg/L ethephon recorded lower acid invertase activities in the bottom internodes than that in the control.

Preservation in vitro of medicinal plant Salvia prionitis

2016/1/15 21:08:18

Effects of the culture mediums(MS,1/2MS and 1/4MS),concentration of sucrose and plant retardants(CCC,PP₃₃₃,ABA,MH)on preservation in vitro of Salvia prionitis were studied. The results showed that 1/2MS culture medium was more suitable for the preservation in vitro of S.prionitis,in this culture medium,plantlets could live a long time with high survival rate. Plantlets grew vigorously and tended to senesce prematurely in the culture medium with sucrose, but had a longer living time with bad morpha and color in the culture medium without sucrose. All of the retardants could retard the growth rate and prolong the preservation time of S.prionitis. CCC at 1.2～1.6 mg/L,PP₃₃₃ at 2.0 mg/L,ABA at 0.5～4.0 mg/L and MH at 0.5 mg/L were favorable for long-time preservation of S.prionitis. While in MS+ABA0.5～4.0 mg/L,the plantlets grow best,in MS+CCC1.2 mg/L and MS + CCC1.6 mg/L culture medium,the plantlets could live for 360 d with 90% survival rate.

In vitro culture of different explants from Pogostemon cablin

2016/1/15 21:08:18

The leaf segments, nodular stem segments,stem segments and root tips from Pogostemon cablin were cultured in vitro. The factors affecting plant regeneration were studied. The results were as follows:BA was effective to induce shoot formation; the optimal BA concentration for shoot formation was 0.1～0.5 mg/L; there was significant difference on the shoot formation from different explants; the leaf segments and nodular stem segments were the most suitable for culture,with the highest shoot formation frequency of 100%; nodular stem segments oriented with their apical ends protruding from the medium could produce more shoots in less time than when they were placed with their basal end upright or were placed horizontally. Optimal rooting medium for regeneration shoots was MT+IBA 0.2 mg/L.

Cloning and idientifcation of β-glucosidase gene of uncultured microorganisms from biogas digester

2016/1/15 10:58:30

The metagenomic DNA of uncultured microorganisms from biogas digester was extracted by indirect extraction,end-blunted and ligated with cosmid vector of pWEB:TNC. A metagenomic library containing 30 000 clones was constructed, the stochastic Analysis of the library indicated that: the average size of the foreign DNA fragments was 40 kb and the capacity of the DNA cloned in the library was 1.2×10⁶ kb. One of the positive clones pGXN100 that show black hydrolysis circles in Esculin plates was subcloned,sequenced and analysed. The results showed that the putative β-glucanase gene had a 1 863 bp ORF,encoding a protein with 620 amino acids sharing 76% and 85% identity with another β-glucanase gene AN292 for DNA and protein levels respectively. The structure functional domain analysis of Unglu100 using SMART software showed it maybe one of beta-glucoside specific PTS system transport proteins.

Studies on the kinetics of cell growth and nutrient consumption in cell suspension cultures of Corydalis saxicola

2016/1/15 10:58:30

The kinetics of cell growth and nutrient consumption were investigated in cell suspension cultures of Corydalis saxicola. The results showed that the pH value decreased during the early period of cultivation and increased later. Total carbohydrate and nitrate were assimilated gradually throughout the growth period,and the absorption of phosphate was the fastest. The growth period of cell suspension cultures was around 20 days with the highest dry weigh and fresh weight of biomass on day 18. However,the highest contents of alkaloids were obtained on day 21,with 8.22 mg/L of dehydrocavidine and 4.31 mg/L of berberine. These obtained results indicated that nutrient consumption was deeply related to cell growth and biosynthesis of alkaloids in cell suspension cultures of C.saxicola. Phosphate was absorbed fastest,followed by nitrate and carbohydrate. It was suggested that phosphate and nitrate were the main compounds,which had effects on cell suspension cultures of C.saxicola.

Activity assay of grain starch synthesis key enzymes in wild rice species of Yunnan

2016/1/15 10:58:30

In order to investigate the amylose synthesis mechanism of the three wild rice species originating from Yunnan so as to utilize the good quality trait of the low amylose content of these wild rice species,four key enzymes(ADPG pyrophosphorylase(ADPGP),soluble starch synthase(SSS),granule bound starch synthase(GBSS),starch branching enzyme(Q-enzyme))activities in three wild rice species and four cultivar rice varieties of Yunnan were assayed during grain filling stages. The result indicated that four key enzymes had similar change trend,but had big differences in the activity among the different wild rice species and the cultivars. It was found that GBSS activity assay in wild rice also had positive relation with amylose content. This indicted that in wild rice the seed amylsoe synthesis is also controlled by granule bound starch synthase. It is also found that SSS and Q-enzyme showed opposition change trend in terms of activity during the grain filling stage. It may suggest these two enzymes had some feedback suppression effect on regulation of starch synthesis.

Physiological compare Merremia boisiana with Pueraria lobata under drought stress and rewatering conditions

2016/1/15 10:58:30

The activities of antioxidant enzymes,such as superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA)content and chlorophyll fluorescence parameters were compared between invasive exotic plant Merremia boisiana and native plant Pueraria lobata under drought stress and rewatering conditions. The results showed that SOD and CAT activities of these plants under drought stress were higher than that after rewatering,and M.boisiana had stronger enzymes activities than P.lobata under same treatments. The difference of MDA content was not evident after drought and rewatering treatments in M.boisiana,while MDA content in P.lobata under drought stress was higher than that of rewatering treatments. Moreover,MDA content in P.lobata was higher than that of M.boisiana under different treatments. These results indicated that M.boisiana had stronger antioxidant activities under drought stress,and its membrane lipid peroxidation level was obviously low. As a result,MDA content was evidently negative relative to the activities of SOD and CAT. Chlorophyll fluorescence parameters Fv/Fm,ETR and ΦPSⅡ after rewatering were significantly higher than that of drought treatments,and the changes of the parameters in P.lobata were a little evident than that in M.boisiana. The value NPQ of M.boisiana rose while that of P.lobata reduced after rewatering,and the latter was higher than the former under drought stress. In contrast,M.boisiana was higher than P.lobata after rewatering. Summarily,it was analyzed that M.boisiana possessed stronger physiological adaptability to drought than P.lobata,which was possibly an important invasive characteristic.

Study on inducion and multiplication culture of bulblet from some parts of flower in Lilium

ZHOU Jun-Hui, ZHOU Hou-Gao, Lin Li-Chan, LU Jun-Tu — 2016/1/15 10:58:30

The some parts in flower such as ovary,style and filament as explants were cultured for inducing bulblet in Lilium. The results indicated that the highest bulblet inducing rate was gotten from filament while the lowest from ovary,there were bigger difference of the bulblets from ovary and style,while there were ordinary smaller but uniform bulblet from filament,there was best inducing effect for bulblet when the media added with 1.0 mg/L 6-BA among all concentration. The uniform bulblets were studied for multiplication culture with different media,different concentration. of 6-BA and NAA,and for enlarging culture with different concentration. of sugar,the results indicated that,MS medium+2.0 mg/L 6-BA+0.2 mg/L NAA was the best one for bulblet multiplication,there was obvious effect of bulblet multiplication culture when the media added with glycine,there was the highest effect for enlarging culture of bulblets when the media added with 4%sugar,but with fewer or more concentration.

The cutting propogation of palmleaf raspberry(Rubus chingii)

2016/1/15 10:58:30

Palmleaf raspberry(Rubus chingii)is one of the important medicinal plants in China. The experiments were conducted in Palmleaf raspberry GAP base of Dexing County,Jiangxi Province,China. The effect of ABT-1 and NAA,on rooting of palmleaf raspberry cuttings was studied to provide references to its rapid propagation in large volume. The results showed that the cutting of palmleaf raspberry without treatment was difficult to develop adventitious roots. Callusing was developed only after treating one time. So the twice treatments with 20 d interval were necessary for rooting of the cutting. Compared with the two auxins,the effect of ABT-1 treatment was better than that of NAA and 50 mg·kg^-1 ABT-1 was the best economically. In a serials of soaking time treatments under 50 mg·kg^-1 ABT-1,2 h for the first soaking and 2-3 h for the second soaking after 20 d later were the best procedure to increased rooting percentage. In different cutting materials,the best cutting period is from March to August for softwood,and from March to May for hardwood as well.

Study on tissue culture of Dysosma versipellis

TANG Feng-Luan, LI Feng, HUANG Ning-Zhen, FU Chuan-Ming, ZHAO Zhi-Guo — 2016/1/15 10:58:30

The seeds of Dysosma versipellis were used as explants for tissue culture in different culture media. The results obtained from the present study were as follows:the seed germination rate could reach 72.4% on MS+BA1.0 mg/L+IBA0.5 mg/L+GA₃4.0 mg/L; the medium which was good for the cluster buds induction in seed buds was MS+BA10.0 mg/L+GA₃0.5 mg/L; the medium of proliferation should be MS+BA(8.0～10.0)mg/L+GA₃2.0 mg/L with low concentration of BA or no BA for circulation culture; the ideal medium for root formation was MS+NAA1.0 mg/L+AC0.2 mg/L,and the rooting rate could reach 100%; tender leaves with stalk could be induced for callus and roots in MS+BA1.0 mg/L+2-ip(0.5～1.0)mg/L+NAA0.02 mg/L. The survival rate of the seedlings was about 90%.

Tissue culture and rapid propagation of Lonicera confuse

2016/1/15 10:58:30

The stems with axillary buds of Guangxi Lonicera confusa were taken as explants to build aseptic system. The results showed that the suitable disposal of sterilization was using 75% alcohol to dispose for 25 s firstly,and then using 0.1% HgCl₂(with Polysor6-BAte 80)to dispose for 9min,the minimum rate of pollution was 6.7%; the suitable initial medium to germinate auxiliary buds was MS supplemented with 6-BA1.7 mg/L,germinative rate was 63.3% which was higher than CK 33.3%; the period of subculture was 30 d,the suitable subculture proliferous medium was MS supplemented with 6-BA1.7 mg/L,the buds were strong,and the average of the proliferous buds were 10.7 which was more than CK 9.5.

Technology procedure of Tissue culture and Propagation of Siraitia grosvenorii

2016/1/15 10:58:30

The primarily technology of tissue culture and propagation of Siraitia grosvenorii was reported,contains the condition of tissue culture,confection of medium,selection and sterilization of explant,inoculation and culture,preservation of superior provenance,hardening-seedling and transplanting,plant pack and transportation. And 5 kinds of referenced medium were reported too. The medium of induced stem was MS+BA0.5～1.0 mg/L+IAA(NAA)0.05～0.1 mg/L+sugar 3%+agar 4.5 g/L,pH5.8. The medium of induced stem apex was MS+BA0.5～1.0 mg/L+NAA0.05～0.1 mg/L+coconut milk 100 mL+sugar 3%+agar 4.5 mg/L,pH5.8. The medium of subculture of fasciculate bud was MS+BA0.3～0.7 mg/L+NAA0.05/IAA0.1 mg/L+sugar 3%+agar 4.5 mg/L,pH5.8. The medium of subculture of cutting was MS+BA0.1 mg/L+IAA0.3 mg/L+active carbon 0.07 g/L+sugar 3%+agar 4.5 mg/L,pH5.8. The rooting medium was MS+BA0.07 mg/L+IBA0.15 mg/L+IAA0.1 mg/L active carbon 0.1 g/L+sugar 3%+agar 4.5 mg/L,pH5.8. On the other hand,the cause of ill phenomenon in the process of tissue culture was analyzed,preventive measure was put forward. And feasible condition of hardening-seedling and transplanting was clear,the relevant manage method was formed. The integrated technology procedure of tissue cultured plantlets of S.grosvenorii was established.

Advances in biotechnological research on Litchi(Litchi chinensis)

2016/1/15 10:58:30

The advances in research on in vitro culture,gene cloning and genetic transformation of litchi were reviewed in this paper. Plant regeneration from pollen,leaves and young embryo had already been succeeded. Some useful genes had been cloned and the genetic transformation mediated by particle bombardment and Agrobacterium tumefaciens infection had been constructed. The problems and prospects of litchi biotechnology were also discussed.

Effects of Cd on physiology and biochemistry and leaf ultrastructure of Seleng wormwood

PAN Jing-Xian, DAI Xi-Ling, SONG Lian-Hua — 2016/1/15 10:58:30

The physiology and biochemistry and leaf ultrastructure of Seleng woodworm were discribed in the experiment with the sandy medium polluted by Cd. The plant treated with the Cd concentration over 180 mg/kg,the assimilation of MDA and relative electricity rate and reduction of POD concentration were observed in leaves. Cd treatment was shown to damage the structure of chloroplasts,as manifested by the swelled shape and the dilation of the thylakoid membranes. Moreover,ultrastructure of cell nuclei was damaged in certain degree,such as shape deformation and nuclei distribution unevenness and chromatin gelatinization. But less than 180 mg/kg Cd,non toxicity of Seleng wormwood was showed for almost treatments,so the Cd resistance range of Seleng wormwood could not be over 180 mg/kg.

Influence of the obligate parasite Cuscuta campestris on the leaf stomatal behavior of its host Mikania micrantha

2016/1/15 10:57:49

It has been recognized that the use of the obligate parasite Cuscuta campestris is one of the most effective means of biologically controlling the invasive weed Mikania micrantha. In order to provide some certain morphological evidences underlying the effects of C.campestris on the physiology of M.micrantha,we studied the influence of C.campestris infection on the morphological parameters of leaf stomata of M.micrantha under a scanning electron microscope. The results indicated that infection with C.campestris had significant effects on diurnal stomatal behavior of M.micrantha leaves. Both stomatal length to width ratio and stomatal area of the leaves in the control plants showed a bimodal diurnal pattern,while those for the infected exhibited a mono-peak pattern. The diurnal peak values of stomata length to width ratio and stomatal area of the control leaves appeared at 12 h and 10 h,while the minimum values appeared at 10 h and 14 h,respectively. From 15 h to 16 h,the ratio and stomatal area reached another less peak value. For the leaves of the infected M.micrantha,the mono-peak values of the stomata length to width ratio and stomatal area appeared at 12 h and 14 h,respectively. The results also indicated that infection with C.campestris had no significant effects on the stomatal density of M.micrantha leaves.

Anthocyanin essence of the purple pigment and positive correlation of the anthocyanin content and the total ginsenoside content of the root tuber of Panax notoginseng

2016/1/15 10:57:49

Panax notoginseng is“the first medicinal material”in Yunnan of China and P.notoginseng produced in the Wenshan eparchy of Yunnan is the Genuine Medicinal Material of P.notoginseng. The transverse sections of the root tubers of P.notoginseng range from yellowish white to purple. The root tubers with pure purple occupy about 28.21% of the root tubers researched,and their pericycles,endodermises,cortexes or epidermises are purple. Specific color reactions and UV-vis spectra indicated that the purple pigment of the root tuber of P.notoginseng belongs to flavonoids,probably holding phenolic o-dihydroxyls,excluding carotenoids,chalcones,aurones,isoflavones and catechins. Anthocyanins and/or their aglycones,namely anthocyanidins,underlay the pigmentation of the purple root tuber and other non-red flavonoids function as co-pigments. The average anthocyanin content and the average total ginsenoside content of the root tubers with pure purple are all the highest,next are those of the root tubers with farraginous color of yellow and purple,and the lowest are those of the root tubers with pure yellow,which is consistent with the changing trend of the color hues of the different root tubers. The difference of anthocyanin contents of the root tubers with different colors reaches the most significant level,but the difference of anthocyanin contents does not reach the significant level. Every root tuber contains anthocyanins of different quantity,and along with increase of the anthocyanin content,the purple of the root color basically becomes more and more obvious gradually. The anthocyanin content of the root tuber is positively related with the total ginsenoside content at the significant level and the correlation coefficient(r)is 0.355. This paper can provide a reference for the exploration on the mechanism of the tuber coloration and the identification of the molecular structures of the tuber pigments of P.notoginseng.

Preliminary study on the cell suspension culture of Eucommia ulmoides and secondary metabolite-chlorogenic acid

WANG Ya-Qin, YE Qing-Hua, ZHU Yuan — 2016/1/15 10:57:49

Some major factors affected the cell suspension culture of Eucommia ulmoides and the production of chlorogenic acid were investigated. The results showed that the concentration of chlorogenic acid was the highest on 15th day,sucrose was better than glucose and maltose for cell growth and chlorogenic acid synthesis and the optimum concentration was 35 g/L. The optimal media was MS media and the proper pH value was 5.3. Some exogenous hormones such as 2,4-D and NAA had little effect on chlorogenic acid synthesis,but 1.0 mg/L 6-BA could promote the production of chlorogenic acid.

Effects of long-lasting brassinosteroid TS303 and propyl dihydrojasmonate on enhancing peanut resistance to chilling

2016/1/15 10:57:49

The effects and the action mechanism of long lasting brassinosteroid(coded as TS303)and propyl dihydrojasmonate(PDJ)on peanut resistance to chilling were studied. The enhancement of chilling tolerance was indicated by the reduction of malondialdehyde and electrolyte leakage. Chilling stress decreased superoxide dismutase(SOD)and catalase(CAT)activities and relative water content while it increased the peroxidase(POD)activity and soluble saccharide and proline contents. TS303,PDJ and their mixture TNZ ameliorated the injury caused by chilling stress through preventing decreases in CAT and SOD activities and relative water content by enhancing the increases in proline and soluble saccharide contents. TS303 exhibited better effect than PDJ on preventing decreases in SOD and CAT activities,meanwhile,PDJ exhibited better effect than TS303 on enhancing the increases in soluble saccharide and proline contents,and it might be the difference in action mechanism that resulted in the additive or synergistic protective effect on cell membrane system.

Genetic analysis of meiotic behavior difference in plants from the same population of Zephyranthes grandiflora

2016/1/15 10:57:49

In this paper,the meiotic behavior of microsporocytes and microspore development in Zephyranthes grandiflora(2n=48)were investigated.The results indicated that there was obvious difference of meiotic behavior in Z.grandiflora plants from the same population.In most plants,lagging chromosome,micronucleolus and other abnormal meiotic behavior could be observed,with an average of 14.02% microsporocytes segregating abnormally. Microspores developed normally,but no fertile pollens could be obtained. Form the abnormal meiotic behavior of microsporocyte in anaphase Ⅰ,it could be proved that the Z.grandiflora was a heterozygote of paracentric inversion for the first time. The meiosis of microsporocyte of a few mutant plants showed disorderly greatly,and multi-polar separation and lagging chromosomes could be readily in anaphase Ⅰ,with and average of up to 94.3% abnormal segregating ate.At tetraspore stage,paraspores raised up to 73.4%.It is thought that abnormality of meiosis in Z.grandiflora was ascribed mainly to chromosome structure variation,and the reason of the behavior abnormality of the meiosis in mutant plants was due to gene mutation that controling the spindle formation,in addition to chromosome structure variation.

Tissue culture of medical plant Erigeron breviscapus

2016/1/15 10:57:49

The tissue culture and rapid proliferation techniques of Erigeron breviscapus were studied by using scape,floral disc and petiole as explants. The explants were cultivated in different MS media with different types and concentrations of plant hormones. The main results can be concluded as follows:the scape is the best material for the propagation among the three explants,scape,floral disc and petiole,as it has the highest inductivity,the best growth and it is easy to produce adventitious buds. The suitable phytohormone compositions to induce callus,adventitious buds and roots are MS+BA 1.0 mg/L+IBA 0.05 mg/L+sugar 3.0%,Ms+BA 2.0 mg/L+IAA 1.0 mg/L+sugar 3.0% or Ms+Kt 3.0 mg/L+IAA 0.5 mg/L+sugar 3.0%,and 1/2Ms+NAA 1.0 mg/L+sugar 3.0%,respectively. The mode of plant regeneration of E.breviscapus in tissue culture has also been investigated.

Analysis of the chromosomal location of 45S rDNA in plants by CPD staining together with FISH

2016/1/15 10:57:25

The chromosomal location of 45S rDNA in sixteen plant species belonging to six families was analyzed using combined PI and DAPI(CPD)staining together with fluorescence in situ hybridization(FISH)with 45S rDNA probe. Fifty three 45S rDNA sites in total were detected in the tested species. The 45S rDNA sites occurred mainly in the short arms instead of the long arms,and interstitial and terminal 45S rDNA sites appeared at similar frequency. Secondary constrictions appeared in most interstitial 45S rDNA sites,but the orientation of the cluster of rDNA repeats relative to the secondary constriction differed among these interstitial sites. The chromosomal distribution of 45S rDNA sites in the tested plants could be classified into 12 types based on the differences in the chromosomal arm in which 45S rDNA site reside,the distance from centromere to 45S rDNA site,the formation of secondary constriction,and the orientation of the cluster of rDNA repeats. The interrelation among 45S rDNA site,NOR(nucleolar organizing region),secondary constriction,and satellite was analyzed based on our results and other previous reports.

Expression of AtPSK2 gene in roselle cells

2016/1/15 10:57:25

By using A.tumefaciens-mediated genetic transformation system of Roselle callus,AtPSK2 gene was introduced into and expressed functionally in Roselle cells. The results demonstrated that,by comparison with un-transgenic Roselle cell or callus,the critical plating cell density and critical initiating cell density of transgenic cell line were cut down 60% and 50%,respectively,the growth cycle of transgenic cell in suspension culture was shortened from 16 days to 12 days,and the specific growth rate of transgenic Roselle callus was raised 75%,the content of flavonoid compounds in transgenic Roselle cell line or callus was hardly altered. The obtained transgenic Roselle cell line in this study not only provides an initial material for selection and breeding of Roselle cell line with high-yield anthocyanin,but also gives a useful research model for further investigation in the mechanism of PSK-α action.

Allelopathic effects of rock plant Pogonatherum paniceum on crops

2016/1/15 10:57:25

In the present study,the allelopathic effects of root,stem and leaf aqueous extracts of Pogonatherum paniceum on seed germination and seedling growth in Oryza sativa,Triticum aestivum,Brassica campestris and Lactuca sactiva by the methods of bioassay were tested. The results showed that all aqueous extracts have allelopathy on four receptor plants,increasing with the increase of concentration. The aqueous extracts of P.paniceum inhibited seed germination and root length,and promoted seedling height in principle. The germination speeds of T.aestivum,B.campestris and L. sactiva were significantly inhibited except O.sativa. No significant effects of aqueous extracts on final germination of receptor plants showed that the allelopathy of P.paniceum delayed seed germination of receptor plants. Sensitivity to aqueous extracts was different between development periods in this study,and seedling period was sensitive than germination period. The leaf allelopathy was 2.14 times of root and 2.57 times of stem of P.paniceum,respectively.

First report Emilia sonchifolia wilt caused by Ralstonia solanacearum

2016/1/15 10:57:26

Emilia sonchifolia bacterial wilt was severely occurred in the field in Guangxi,China. The pathogenic bacterium was confirmed by Koch postulate. Then Polymerase chain reaction(PCR)amplification was conducted using 16s rDNA universal primers to identify the pathogenic bacterium. PCR products sequencing results showed the pathogenic bacterium had 99% homology to Ralstonia solanacearum. This is the first report of E.sonchifolia wilt caused by R.solanacearum.

Allelopathic effects of Parthenium hysterophorun aqueous extract on germination of Bidens pilosa and Delavaya toxocarpa

2016/1/15 10:57:26

The seeds of accompanied species Bidens pilosa and energy plant Delavaya toxocarpa were treated with the aqueous extracts of root,stem and flower of Parthenium hysterophorus. The germination rate,index of allelopathic effect and germination speed of seeds were measured. The results showed that aqueous extracts from three parts of P.hysterophorus had different effects on the seed germination of B.pilosa under different concentrations,while that didn't work on D.toxocarpa. This experiment will provide reference basis for choosing feasible plant to against P.hysterophorus by biological control strategy.

Study of latex exploitation using depolymerizing compounds of microfilaments in rubber trees

2016/1/15 10:57:26

Stimulated latex exploitation was investigated in rubber trees with saturated macrolides,KI(1%,5%,10%)and 0.3% ethylene.Samples treated with 2% methyl-cellulose were used as blanks. Latex output and its six physiological parameters,the contents of pure rubber,total solid,sucrose,inorganic phosphorus,thiol and Mg²⁺,were measured at the same time. Results showed that application of 1% KI or saturated macrolides could lead to increase of latex yields of rubber trees. The increasing extents of 1% KI or saturated macrolides treatments were almost equivalent to that of 0.3% ethylene treatment used as a common stimulus in latex exploitation nowadays. There were no obvious diversities in pure rubber content and total solid content in latex from rubber trees treated by 1% KI,saturated macrolides and 0.3% ethylene,except for the other four physiological parameters. In addition,KI of high concentration(5% and 10%)was harmful to bark of rubber trees and even caused fatal tapping panel dryness.

Growth and biomass allocation of Artemisia annua from different seed sources

2016/1/15 10:57:26

In this paper,the growth and biomass allocation of Artemisia annua from Du'an, Chongzuo, Yangshuo and Rong'an were studied. The results are as follows:the growth of A.annua was the fastest from the end of May to early July,the peak of growth was in the middle ten days of June. In this period,more fertilizer should be applied; the mean values of all functional module biomass in different seed sources exhibited same regularity:stem modules>leaf modules>root modules; the seed source of A.annua in Du'an which leaf module biomass and artemisinin content were highest in all seed sources,was excellent.

Effects of different treatments on cuttage cultivation and growth of Keteleeria calcarea

2016/1/15 10:57:26

Keteleeria calcarea is one of the rare and endangered tree species. The stem is tall and thick. Since the flowering and seed setting rate in fruit-bearing shoots are few,seed collecting is very difficult. Moreover,neither the germination percentage of seeds nor the propagation rate is high. In this research,for its convenient and rapid propagation,cuttage seedling was conducted through difference treatments by comparison analysis respectively. Three factors,including rooting agent,fertilization condition and cutting diameter on seedlings raising and growth of K.calcarea,which is close related with survival rate,was discussed. The result shows: treated with ABT rooting powder at concentration of 150 mg/L,while cutting diameter is 0.70 cm,the average percentage of survival represents 95.6%. After that,manure and carbamide(manure:carbamide=100:0.4)are optimal fertilizer on cutting to meet the growth requirements of robust shoots. The average shoot growth is up to 30.8 cm.

Peroxidase isozyme in sprouting seeds of genus Fagopyrum

2016/1/15 10:57:26

The peroxidase isozyme of thirty-two collections of cultivated and wild buckwheat belonging to nine species of genus Fagopyrum including eight species of the big-achene group and one species of the small-achene group were studied by means of polyacrylamide gel eletrophoresis(PAGE). The results showed that there are twenty-three different bands of peroxidase isozyme,with a range from four bands to eight bands among different buckwheat species. F.esculentum and F.tataricum have seven bands and four bands among different collections,respectively. The variations of peroxidase isozyme zymograph are very great among different buckwheat species but less in the same species. The results of the zymograph analysis and the clustering showed that there are much great difference of zymographs between the big-achene group and the small-achene group and that F.megaspartanium and F.pilus are close in clustering tree to F.esculentum and F.tataricum,respectively,supporting Chen's hypothesis that F.megaspartanium and F.pilus may be the ancestors of F.esculentum and F.tataricum,respectively.

Physiological and biochemical characteristics of Myricaria laxiflora,an endangered species in the Three Gorges Reservoir Area

2016/1/15 10:57:05

In order to discover the physiological and biochemical characteristics in Myricaria laxiflora,an endangered species in Three Gorges Reservoir Area,the transpiration,photosynthesis,water potential of plant planted in two kinds of soils were tested in different growing seasons. Results showed that changes of transpiration,photosynthesis and water potential in a day all showed “single curve”. Transpiration and photosynthesis could reach the maximum of a day at noon whereas water potential fall to the minimum. Transpiration changed with the fluctuation of photosynthesis. The photosynthesis and transpiration of plants in May were higher than that in October due to the plants in October just recover from the dormancy induced by flooding. PV-curve and relative water parameters,including Ψ₁₀₀(maximum osmotic potential at water saturation point),Ψ₀(osmotic potential when a plasmolysis just emerge),ROWC(relative osmotic water content),RWC(relative water content),AWC(relative water content in apoplast)and RDW(relative dry weight)of the plants grown in sandy soil,red soil and submerged soil were measured. Plants at submerge sandy soil having low Ψ₁₀₀,Ψ₀,AWC and high ROWC and RDW implied that submersion could promote plants to develop a good ability resisting dry stress. The biochemical contents related with stress resistance were tested in different growing seasons. Mlondialdehyde(MDA)content,peroxidase(POD)activity,Polyphenol oxidase(PPO)activity and total soluble sugar content decreased after flower and fructification. It suggested that the plant resistance declined after fruit. Some suggestions finally were put forward for the population transplant to conserve this species according to its physiological and biochemical characteristics.

Regulatory effects of gibberellic acid(GA₃) on shoot and root formation of Huang-qin (Scutellaria baicalensis)in vitro

2016/1/15 10:57:05

Influence of exogenously applied gibberellic acid(GA₃)on shoot and root formation and development in shoot tip and stem tissue cultures of Huang-qin(Scutellaria baicalensis),an important medicinal plant,was investigated. GA₃ application at the concentrations of 1-20 μmol/L resulted in the significant enhancement of shoot formation and growth while it reduced root formation and development. In different transferring tests,the exogenously applied growth regulators GA₃ and IAA resulted in significant changes in the concentrations of major flavonoids in Huang-qin tissues. The highest concentrations of baicalin, baicalein and wogonin were 14.90,2.70 and 0.54 μg·mg^-1(dry weight),respectively,when the explants were pre-cultured on 2.5 μmol/L IAA medium for 6 days before transferred to MSO medium containing GA₃ at 5 μmol/L in continuous darkness.

Inhibitory mechanism of extracts from Ginkgo biolaba sarcotesta on mushroom tyrosinase

2016/1/15 10:57:05

Three extractants(water,ethanol and ethanol-aether mixture)were isolated from Ginkgo biolaba sarcotesta,and then three kinds of extracts were used to test the inhibition on the activity of mushroom tyrosinase. The results showed that three extracts could inhibit the activity of tyrosinase obviously. The inhibitor's concentration leading to 50% enzyme activity lost,IC₅₀ was estimated to be 2.25,1.75 and 0.32 mg/mL,respectively. Their inhibition mechanisms were analyzed. All of three extracts showed reversibly inhibitors to tyrosinase and the type of inhibition belonged to be mixed-type. The inhibition constants,K_I and K_IS,of each extract were determined respectively and compared. The value of K_I was determined to be 2.11,1.62 and 0.29 mg/mL,respectively; the value of K_ISwas determined to be 2.80,2.33 and 0.45 mg/mL,respectively. The ethanol-aether extracts showed the best inhibition on tyrosinase.

Analysis of some quality characteristics in soybean mutants

2016/1/15 10:57:05

In order to gain the new soybean variety strains,it is necessary to improve the existent soybean resource to increase the soybean variety and improve the soybean qualities. After the soybean variety DongNong 42 which has more protein than others and the DongNong 163 which have more fat than others were treated with NaN₃,32 mutants from DongNong 42 in its M₆ generation and 54 from DongNong 163 were selected,and their agronomic characteristics(high,hundred grains,etc.)and their qualities(fat,water-soluble protein and alkali dissolved protein)were analyzed. The result indicates that:(1)the coefficients of variation of all strains'quality character are different,so it is effective to select qualitative character from the descendants;(2)the content of fat and content of water-soluble protein are remarkably negative correlation among variation strains.

Cloning and analysis of hmgr gene conserved fragments in Descurainia sophia

2016/1/15 10:57:05

Based on the design of degenerated oligonucleotides according to the conservative regions of eight 3-hydroxy-3-methylglutaryl-CoA reductases(HMGRs)from six plants and total RNA extracted from Descurainia sophia,a 458bp fragment of hmgr was obtained by using reverse transcription PCR strategy. Through sequence analysis by BlastP online,the resulting protein showed high homology to 3-hydroxy-3-methylglutaryl-CoA reductase,with 98% identification compared with Arabidopsis thaliana(NP_177775),96% with Raphanus sativus(CAA48610),88% with Eucommia ulmoides(AAV54051),89% with Picrorhiza kurrooa(ABC74565),86% with Camptotheca acuminata(AAB69726)and 87% with Gentiana lutea(BAE92730). Deduced amino acid sequences were also analyzed by PROSITE,ClustalX and MEGA Ver.3.1,and data present evidence for the existence of 3-hydroxy-3-methylglutaryl-CoA reductase in Descurainia sophia. This is the first report of the hmgr gene isolated from D.sophia.

Purification and kinetic characteristics of an acid phos- phatase isoform from germinating pigeonpea seed

2016/1/15 10:57:05

Using p-nitrophenolphosphate(pNPP)as substrate,one isoform of acid phosphatase from germinating pigeonpea seed,encoded as APase Ⅱ,was purified to 247 folds and the specific activity 51.8 U/mg protein through ammonium sulfate fractionation and three sequential DEAE-Sephadex A 25,Hydroxyapatite and Concanavalin A-Sepharose 4B column chromatography. The purified APII was demonstrated by Native- and SDS-PAGE to be electrophoretic homogeneity and as a 33 kDa monomer. APase Ⅱ exhibited optimal pH at 5.0 and optimal temperature at 35 ℃ and strong stabilization at the pH ranging from 3.5 to 7.0 and at temperature below 55 ℃. APase Ⅱ showed a highest specificity against pyrophosphate,and was activated by K⁺ and Mg²⁺,while inhibited by Fe²⁺,Mn²⁺,Mo₇O₂₄₆^-,F- as well as by organic acids including as tartrate,malate,isocitrate,oxalate,citrate,glycolate,glyoxylate and ascorbate.

Existent states of anthocyanins in vacuole and their coloration effects in higher plants

ZHAO Chang-Ling, ZHANG Li-Mei, LIU Fu-Cui — 2016/1/15 10:57:05

This review sums up the reasons of anthocyanins being sequestered into vacuole,the existent states of anthocyanins in vacuole and the corresponding coloration effects of anthocyanins on plant cells. Transporting of anthocyanins from the biosynthesis site,namely the cytoplasm of plant cell,into vacuole is to detoxify the toxicity of anthocyanins on the functional molecules of the cell,such as proteins and DNAs. The vacuolar compartmentalization of anthocyanins is the prerequisite for anthocyanin function normally in plant cells. In a wide range of plant species and in most cases,anthocyanins dissolve completely in vacuole. However,in vacuoles,anthocyanins can also form granules which can be classified into two categories,namely anthocyanoplast(ACP)and anthocyaninic vacuolar inclusion(AVI). ACP is membrane-bounded,its formation is the result of the progressive coalescence of the smaller pigmented vesicles in vacuole and fully developed ACP is typically spherical and more deeply red-colored than the vacuole. In vacuole,ACP is high density and insoluble globule highly concentrated with anthocyanins. The emergence of ACP can provide intense coloration in the vacuole. AVI may be protein matrix and it possesses neither a membrane boundary nor an internal structure,its formation is the result of the anthocyanins transported into the vacuole bind with a protein matrix. In vacuole,AVI is irregular and jelly-like in shape. In AVIs,the attachment of anthocyanins to the matrix protein is likely to be via H-bonds to a sterically restricted site. AVI is suggested to act as vacuolar anthocyanin “trap”,preferentially for anthocyanidin 3,5-diglycosides or acylated anthocyanins. The emergence of AVI can enhance color intensity and results in the “blueness” of color in the vacuole.

A preliminary study on the water physiology of four plant species in karst area of Southwest China

2016/1/15 10:57:05

The water characteristics of Alchornea trewioides,Sapium rotundifolium,Pteroceltis tatarinowii and Sinoadina racemosa were measured and compared in the paper. The results showed that the tissue water content,critical saturation deficit and the water requirement degree were significantly different among the examined four plant species. The drought-resistance of Pteroceltis tatarinowii,a deciduous shrub,was the strongest among them,Alchornea trewioides had the highest WUE saturation point of 600 μmol·m^-2·s^-1,the PAR was logarithmly correlated with stomata conductance; the LSP of water use efficiency of the three deciduous trees species was all at 400 μmol·m^-2·s^-1. The PAR was significantly correlated with transpiration and the stomata conductance,and no transpiration saturation phenomenon in the four plant species was observed.

Influences of different gibberellin and media on growth of seven cultivars of colour callalily

ZHAO Jian, ZHANG Cui-Ping, LI Xiu-Juan, QIU Shuo — 2016/1/15 10:57:05

Gibberellin accelerating germination to 7 cultivars of colour callalily and different media on its growth were researched in the paper. It was shown that the germination of 7 cultivars of colour callalily appeared to increase initially and then reach their peaks at 1 000 mg·L^-1 and tended to decline later on with GA₃ treatments, and it was higher significantly that the germination of 7 cultivars of colour callalily by 1 000 mg·L^-1 GA₃ than that of CK(P<0.05). In the 3 media, it was better that the height, bulbs weight, flowering stage, length of flower stems and spathe diameter of 7 cultivars of colour callalily which grew in the medium 2 than those grew in other 2 media. 08,21,33,37 and 47 can be cultivated in pots and cuting flowings,but 09 and 16 only can be cultivated in pots.

Isolation of protoplast from the Astragalus membranaceus

2016/1/15 10:57:05

Leaves and callus of the Astragalus membranaceus were used to examine the conditions for protoplast preparation and culture of the A.membranaceus. The results showed that the leaf segment of the A.membransceus could release massive and vigorous protoplasts. Compared with the callus of the species,the protoplasts extracted from the leaf segment are more efficient in the cell fusion and tissue culture. The optimized treatment is 2% cellulase,0.5% hemicellulase,and 0.5% pectinase for 12 h.

Electrophysiological mechanism of root length growth of Arabidopsis thaliana inhibition by JA-Me

2016/1/15 10:57:05

Arabidopsis thaliana(ecotype Columbia)was treated with methyl-jasmonate(JA-Me)and the relations of JA-Me,H₂O₂ and inward potassium channel were examined by using patch clamp technique. The results showed that different concentrations of JA-Me(10^-8-10^-3mol/L)could all inhibit the elongation of primary roots to some degree. The inward K⁺ currents of root cortical protoplasts were inhibited with 10^-4 mol/L JA-Me and 10^-5 mol/L H₂O₂ treatment. What's more,10^-4 mol/L DPI and 10^-2 mol/L Vc could both reverse the JA-Me-inhibited inward K⁺ currents respectively,which suggested that H₂O₂ may act as a second messenger involving JA-Me inhibition of the root inward K⁺ currents. The results indicate that JA-Me inhibition of root length growth may result from the H₂O₂ induced JA-Me inhibition of the root inward potassium channel.

Received date: 2006-05-17 Accepted date: 2007-10-16
Foundation item: Supported by the National Natural Science Foundation of China(30270924); Natural Science Foundation of Anhui Province (070411004); the National Science Foundation of Education Department of Anhui Province(2006KJ184B); the Foundation of Provincial Key Lab of Biotic Environment and Ecological Safety in Anhui Province
Biography: XIAO Jia-Xin(1970-),male,Born in Luotian County of Hubei Province,Associate professor,Ph.D,mainly study on physiology and ecology of horticultural plants. Annual changes of iron,manganese,zinc and copper concentrations in both types of citrus fruit

2016/1/15 10:56:00

Annual changes in concentrations of iron(Fe),manganese(Mn),zinc(Zn)and copper(Cu)were measured in whole fruits from parthenocarpic Kamei satsuma mandarin(Citrus unshiu)and self-pollinated Egan 1 tangerine(C.reticulata)trees. The results were showed as follows:(1)Zn and Cu concentrations in the ovary of Kamei were relatively high before flowering and at full bloom,and decreased after flowering,whereas those of Egan 1 decreased obviously and were relatively low at full bloom,thereafter significant increases were observed. There were no significant differences in changes of Fe and Mn concentrations in the ovaries(fruitlets)between the two cultivars,which presented similarly decreasing trends after flowering.(2)Fe,Mn,Zn and Cu concentrations were relatively high in whole fruits of both cultivars during young fruit development,and decreased remarkably during early fruit enlargement(drought spell),whereas increased dramatically at the middle stage of fruit enlargement,thereafter decreased gradually. Dynamics of micronutrients concentrations in developing fruit and their possible relation with fruits development is discussed herein.

Physiological mechanism in bitter melon infected with powdery mildew fungus

WANG Guo-Li — 2016/1/15 10:56:00

The study focused on the pathogenic process of powdery mildew and the physiological mechanism in Dading bitter melon. The result showed that the membrane permeability increased less compared with the control in pathogenic leaf,and its change wasn't obviously related to the pathogenic index. The contents of chlorophyll a,chlorophyll b,total chlorophyll and carotenoid all decreased gradually along with the pathogenic process developed in pathogenic leaf,which decreased by 72.6%,68.5%,1.5% and 27.1% respectively compared with their controls after the plant was inoculated for 15 d. Among these changes,the decreases on chlorophyll a,chlorophyll b and total chlorophyll all presented high relativity to pathogenic index,and the highest relativity was found in chlorophyll a. The test result of oxidases indicated that POD and PPO activities both increased compared with the control in pathogenic leaf,and increased more along with the pathogenic process developed,which could increased by 528.5% and 70.4% respectively compared with their controls after the plant was inoculated for 15 d,and the change on two oxidases activities showed high relativity to pathogenic index.

Changes of superoxide dismutase and estrase in processes of embryogenic calli induction of Eustoma grandiflorum

LIU Ming-Zhi, CHEN Qin, LIU Xi — 2016/1/15 10:56:00

Embryogenic calli induction of Eustoma grandiflorum were carried out when non-embryogenic calli on necSM(MS medium containing 1.0mg/L 2,4-D and 0.5mg/L KT)were transferred onto ecIM(MS medium containing 1.0mg/L 2,4-D). Changes of superoxide dismutase(SOD)activities and isoenzymes and estrase(EST)isoenzymes in calli of E.grandiflorum cultured on ecIM and necSM for different days of cultures have been achieved. Experimental results have been shown that SOD activities in calli of E.grandiflorum on ecIM and necSM were low in early culture times and raised along with increasing days of cultures and maintained a higher level without an evident regularity for the changes of SOD activities,and for SOD isoenzymes there was also a new band of SOD isoenzyme until 4 days of culture; Estrase(EST)isozyme in calluses of E.grandiflorum cultured on ecIM and necSM had an distinct absence in the periods from 16 days to 20 days.

Effect of different wavelength lights on photosynthesis and growth of Siraitia grosvenorii

2016/1/15 10:56:00

The test-tube plantlets of Siraitia grosvenorii were cultivated under different wavelength lights(blue light 475±5 nm; yellow light 585±5 nm; red light 660±5 nm and white light). Growing condition was observed after 25 days. The morphological characters were arranged:blue light>white light>red light>yellow light. Weight of single plantlet was:blue light>red light>yellow light>white light. Under blue and white lights,seedlings have large and deep green leaves,short and thick stems with more lateral buds. While under red and yellow lights,the leaves were small and yellow green; the stems were long,crook and thin. Based on the absorption spectrum of its ripe leaves,two strong absorption areas were found in 380～500 nm and 660～680 nm. The photosynthetic rates under different lights were in the order:red light>blue light>white light>yellow light. All these results indicated that blue light was best for growth and development of young seedling of Siraitia grosvenorii. While the red and blue lights were suitable for photosynthesis.

Endophytic fungi screening from Atracty lancea and inoculating into the host plantlet

2016/1/15 10:56:00

The endophytic fungi were screened and identified from Atracty lancea. Sixteen strains of endophytic fungi were screened totally,fourteen from leaves. One strain was from root and stem respectively. These strains including Alternaria sp.,Fusariium sp.,Cunninghamella sp.,Penicillum sp.,Phoma sp.,Sclerotium sp.,Gilmaniella sp.and Myceliasterilia. One strain of Sclerotium and one strain of Gilmaniella which no harm to plant tested by culturing together with tissue rapid propagation plantlets were inoculated into propagation plantlets. The influence of the endophytic fungi on the growth,livability and physiology of A.lancea were studied. Sclerotium sp.can strongly increase the livability of A.lancea. The mycelium could be observed by dying the material. Superoxide dismutase and peroxidase activilities of treated groups were both higher than that of control. Index of unsaturated fatty acid was the same as that of control. These were consistent with the result that endophytic fungi were symbiosis with the host.

Studies on several physiological and biochemical indices of the heat-resistance of Lilium longiflorum and its comprehensive evaluation

2016/1/15 10:56:00

The heat-resistance comprehensive evaluation of eight genotypes of Lilium longiflorum were studied on the basis of the variety of four physiological and biochemical indices such as the relative water content,the proline content,the soluble protein content and the relative electronic conduction rate under different temperatures stress(25 ℃,38 ℃,42 ℃,46 ℃,50 ℃). The results showed that the heat-resistance difference among genotypes was significant. The order of heat-resistance among L.longiflorum genotypes was K_1-1,K_2-7,K_1-2,F₁,K_2-2,Wforest,G,Wfox.

Glands structure of liquorice and its regulation mechanism on osmosis stress

WEI Sheng-Lin — 2016/1/15 21:07:58

Foliar glands of liquorices are special morphological structures for drought tolerance. The glandular organ regulates osmotic potential of diachyma cells by secretes polysaccharide,and it's also an important physiological characteristic of drought tolerance. During folded period,the two sides of young leaves are symmetrical,and about 13.0% of glandular organ heads start to secrete or accumulate polysaccharide liquid ball. About 11.6% glandular organ starts to secrete polysaccharide liquid. About 75.3% glandular organ doesn't secrete polysaccharide liquid. It's indicated that some glandular organ starts to take part in osmoregulation at that time. All glandular organ heads of mature leaf secrete polysaccharide liquid ball which indicate that all glandular organ participate osmoregulation by secrete polysaccharide toward outside. Compared with the control,the level of leaf polysaccharide increased 59.8%(P<0.01)when osmosis regulation cultivates with 15% PEG+Hoagland. Liquorices leaf secrete polysaccharide liquid toward outside in order to keep suitable osmotic potential by glandular organ,which is an initiative adjusting process and style.

Effect of Cadmium on physiological characteristics of mangrove Avicennia marina and Aegiceras corniculatum seedlings

SHI Gui-Yu, KANG Hao, DUAN Wen-Fang — 2016/1/15 21:07:58

Based on the pots experiments in sand and mud culture,the effect of heavy metal Cd on some physiological characteristics of Avicennia marina and Aegiceras corniculatum mangrove seedlings were studied when treated by saline water with different concentrations of Cd(0.6-30 mmol·L^-1). The results indicated that at the concentration of 0.6 mmol·L^-1,the chlorophyll content and the photosynthetic rates of Aegiceras corniculatum and Avicennia marina seedlings remained unchanged,the mangrove seedlings also grew well. When Cd concentration was higher than 12 mmol·L^-1,the leaf chlorophyll content and activities of POD and SOD in Avicennia marina and Aegiceras corniculatum seedlings began to decrease. But as the increasing of the concentration of Cadmium the activity of POD got lower and lower. The results were showed that Cd had poisoning effect to mangrove; higher Cd concentration could lead to more serious poisoning effect. Comparing the two terms,Aegiceras corniculatum had a higher tolerance to Cd than Avicennia marina.

Effect of aluminium on protection enzyme system and proline of Brugiera sexangula seedlings

2016/1/15 21:07:58

The activity of POD,CAT,SOD,APX,the content of soluble protein,MDA and proline in the leaves and roots of Brugiera sexangula were studied under 10-50 mmol/L AlCl₃ treatments. The results showed that,B.sexangula had strong tolerance to high concentrations of Al³⁺,even 50 mmol/L of Al³⁺,but under this concentration,the membrane system in the leaves and roots were seriously damaged due to peroxidation,while the content of MDA increased and the active oxygen metabolism of the cells were unbalanced. In protection enzyme system,Al³⁺ treatments concentrated on the raising of APX and POD activity,reducing of CAT and SOD activity in leaves; for the roots,the POD and SOD activity were significantly increased,while the CAT activity were decreased. Under treatments of 25-50 mmol/L Al³⁺,in both leaves and roots,the soluble protein content decreased evidently but the Pro content increased.

Analysis on the different characteristic of calcium in the seed of Cycas guizhouensis

2016/1/15 21:07:58

The total calcium and the content of characteristic of calcium variation in different tissues of fresh ripe seeds of Cycas guizhouensis are determined,then the store mechanism and the distribute regularity of calcium in C.guizhouensis seeds during its growth are explored. The results showed that the total calcium in fresh ripe seeds of C.guizhouensis were decreased with the order: episperm>mesocarp>embryo; the mainly variation of calcium HCl-calcium in episperm,which are 470.8 mg/kg,accounted for 56.3% in emisperm,24.6% in tissue,and 11.4% in entire seed,respectively,it's play important roles of fountain and warehouse during the growth of embryo; the mainly calcium variation in mesocarp and embryo was organic soluble calcium,which are 70.2 mg/kg in mesocarp and 70.6 mg/kg in embryo,and the organic soluble calcium have position function in building the structure of seeds and in the growth of embryo. Additionally,the content of soluble protein and soluble sugar in the seeds of C.guizhouensis had closely relation to the higher calcium content and higher calcium variation content,and the accumulation of characters of macromolecules had position significance for the C.guizhouensis to adapt the higher calcium environment.

Advances on screening of Cd-hyperaccumulators and transgenic improvement of plant in Cd-accumulation

2016/1/15 21:07:58

Phytoextraction is an economic and environment-protected remediation technique of Cd-contaminated soils. The key is to screen Cd-hyperaccumulators or enhance the ability of Cd-accumulation by genetic engineering. Seven Cd-hyperaccumulators and some potential Cd-hyperaccumulators have been discovered. In addition,genes related with Cd accumulation in different organisms were studied. These genes included:(1)genes encoding Cd accumulation related enzymes,such as ACC of bacterial,PC synthase of plants;(2)genes encoding metal-chelatin,such as MT(Metallothionein),transport protein;(3)other genes,such as Hvhsp17,PvSR2(Phaseolus vulgaris stress-related gene number2). Some of these genes have been transformed to plants and microorganisms to enhance their cadmium tolerance and accumulation,which provides a new way for realizing the aim of remediation of Cd-contaminated soils.

Construction of a plant expression vector containing antifreeze protein(afp)gene and it's transformation into banana(Musa AAA Cavendish) embryogenic cell suspension

2016/1/15 21:07:58

Antifreeze protein(afp)gene was amplified by polymerase chain reaction using genomic DNA extracted from the seedling of carrot cv.Jingdou Qicunrenshen. Sequencing result showed that there was no difference in nucleotide sequence between this afp gene and that cloned from‘Wuzhong'carrot in Ningxia. afp gene was subsequently cloned into intermediate vector pMD18-T and binary vector pBI121 to construct plant express vector pBI121-afp,which was transformed into Agrobacterium tumefaciens strain EHA105 by freeze-thaw method. Agrobacterium tumefaciens-mediated method was used to introduce afp gene into the embryogenic cell suspension of Musa AAA Cavendish cv.Beida Aijiao,followed by plant regeneration via embryogenesis. PCR analysis showed that 2 from 9 obtained regenerated plants resistant to kanamycin were positive,which possibly indicated that afp gene has been integrated into the genome of banana plants.

Construction and transformation of antisense RNA expression vector of Arabidopsis phytochrome A gene

2016/1/15 21:07:58

Phytochrome A(PhyA)is the specific photoreceptor which receives far-red light in plants. Many critical genes,including auxin response factor 8(ARF8),are regulated by PhyA. Therefore,it is necessary to construct the PhyA-antisence transgenic lines for revealing the mechanism underlying phyA-regulated genes expression. The PhyA CDS was amplified from Arabidopsis total RNA by RT-PCR method. Then,it was inserted reversely into pMD1 to get the antisence expression vector pMD1-PhyA CDSR. The transgenic lines were acquired by agrobacterium-mediated method. 13 transgenic lines have been acquired by Kmr screening and PCR analysis. The vector pMD1-PhyA CDSR and the transgenic lines will be useful to study how the genes,such as ARF8,are regulated by PhyA.

Studies on pollen germination and storage of Cycas species

2016/1/15 21:07:58

Pollen suspension culture was conducted under combinational conditions of different grades of pH and concentrations of sucrose and boric acid to estimate pollen viability of Cycas debaoensis,C.bifida,C.parvulus,C.elongata. Pollen viability of C.parvulus and C.elongata was also estimated after being stored in different temperature and humidity conditions. The results showed that:(1)the optimal medium for the germination of those cycad's pollen was compose of 1%-2.5% sucrose+100-500 mg/L boric acid and pH was in the range of 6.5-7.0;(2)the pollen stored in a sealed container with desiccant in room temperature was viable up to one month;(3)the pollen being stored under 0 ℃ in a container without desiccant could kept its viability over 4 months;(4)through artificial pollination,the seed setting rate of C.elongata could be up to 90.3% by using liquid nitrogen stored pollen,which is no remarkable difference to those of using fresh pollen;(5)the pollen of C.elongata,after stored in liquid nitrogen for period of time,did not show significant loss of viability after dried to 13.2%-15.5% of water content and the results indicated that stored in liquid nitrogen can be used as a long term storage measure for cycad pollen.

Study on in vitro selection of Pogostemon cablin resistant to bacterial wilt

2016/1/15 21:07:58

The leaf segments and nodular stem segments from Pogostemon cablin were cultured in vitro. The factors connected with the in vitro selection,such as the different antiseptic methods of R.solanacearum crude toxins,the cultural time and the concentrations of R.solanacearum were studied. The results showed that:The survival rate of explants on the media with R.solanacearum crude toxin sterilized from filters is lower,which showed the toxicity of the crude toxin sterilized from filter was stronger than that of crude toxin sterilized in hot and damp. The explants on the media with the crude toxin from cultured 12 h bacteria were prominently browning and dead. And after 30 d,the shoot formation rate of leaf segments and nodular stem segments were 10.33% and 36.11% respectively. Crude toxin from bacteria concentration above 1.41×10⁸ cfu/ml could inhibit the regeneration of explants obviously,with high death rate and low shoot formation rate. And also,the root formation of the shoots from the explants was difficult at that concentration of crude toxin. The R.solanacearum crude toxin had toxicity to the explants and could inhibit the plant regeneration of P.cablin. The system of in vitro selection of P.cablin resistant to bacterial wilt was established

Screening a group of cellulose-degradation bacteria and their zymogenic characteristics

2016/1/15 21:07:58

Bacterial strains were isolated from humus mud by employing the dilution methods. Seven cellulose-degradation strains marked as B-37,B-35,B-31,B-25,B-17,Z-a,and Z-b were selected according to the hyaloid rings on the medium of carboxymethylcellulose sodium(CMC-Na)- Congo red and their efficacy of decomposing filter paper. Z-b had the strongest ability in decomposing filter paper among the seven strains and 16S rDNA analyzed indecated that it was Stenotrophomonas maltophilia owning to 99.8% homology. Comparing clony size of the seven strains in media with different carbohydrates and inorganic salt,we found that the potato extract and inorganic salt which contained CaCl₂ 0.20,MgSO₄ 1.25,NaCl 5.00,(NH₄)₂SO₄ 1.30,KH₂PO₄ 1.35,FeSO₄·7H₂O 0.015,Na-EDTA 0.02(g/L)were suitable for the bacteria growth. Studying cellulase activity of 7 strains in different cultivation conditions,it's found that Z-b had the hightest filter paper(FPase)and CMC(CMCase)activity in the media which filter paper as the only carbohydrate. They were 0.099 and 0.075 U/mL. But on PSA medium,it was B-31and B-37 getting a highest cellulase activity. They were 0.131 and 0.175 U/mL. When fermented individually with filter paper,only Z-b could decompose the whole pieces of paper into powders. But when fermented with fungus 34,the combinations of Z-a+34,Z-b+34,B-31+34,and B-25+34 could hydrolyze the whole filter paper into water-soluble compounds. It was obvious that the cellulase activity of the seven strains closely related with their cultivation condition. Cooperation exists between some fungi and bacteria,it could greatly improve the efficiency of cellulose hydrolyzation when mixed fermentation.

The optimization of fermentation conditions of Candida sp.GXU08 lipase

2016/1/15 21:07:59

The synthesizing of cyclopentadecanolide(a kind of macrocycle musk)by Candida sp. GXU08 lipase has been concerned. Under certain conditions,the converse of cyclopentadecanolide was directly connected with lipase activity. The higher the lipase activity was,the stronger the conversions were. The fermentation conditions of Candida sp.GXU08 lipase were optimized through the single factor and the orthogonal experiments. The results showed that:The compositions of optimal fermentation medium were sucrose 0.5%,starch 0.5%,peptone 1.5%,K₂HPO₄ 0.05%,MgSO₄ 0.15%,(NH₄)₂SO₄ 1%,camellia oil 1.5%,rapeseed oil 1.5%,pH=8.0 Under the fermentation condition of 28 ℃,180 r/min,48 h,the lipase activity would reach 27.53 U/mL,which was 3.74 times of the original culture medium. The conversion of cyclopentadecanolide was 16.6%,which was 4 times of unoptimized.

Studies on the antioxidant activity of Coptis chinensis

2016/1/15 21:07:59

Antioxidant activities of different extracts from different parts of Coptis chinensis were evaluated by the DPPH assay,FRAP assay and ABTS assay,and the results were compared to Trolox. The results showed that different parts had different activity,and fibrous root had the highest antioxidant activity; and different polar solvent extracts of the same part also demonstrated different antioxidant activity,ethyl acetate and methanol extracts generally had higher antioxidant activity than the petroleum ether extract. The antioxidant activity of the extracts from different parts of C.chinensis increased when the concentration of the extracts increased; there were good correlation among the three methods,and the values obtained by FRAP assay were correlated best with those obtained by DPPH assay(r=0.9261,P<0.01).

Contents of effective components in flowers of Eriobotrya japonica

2016/1/15 21:07:59

With the ultraviolet spectrophotometry and HPLC, the contents of total flavonoids and triterpenoid in loquat flowers from different localities and growth years were studied for determining the optimum localities and the changes of effective component contents in different growth years. The results showed that the differences of total flavonoid and triterpenoid contents among the samples of different localities were significant. The total flavonoids contents of samples from Longquan,Renshou,Panzhihua and Putian were higher than that from other localities(P<0.01). The oleanolic acid content of samples from Longquan,Renshou,Pujiang and Putian were higher than that from Suining and Neijiang(P<0.05 or P≤0.01). The ursolic acid content of samples from Yuhang was the highest,which was higher than that from Suining,Panzhihua,Bishan and others(P<0.01). In different growth years the contents of effective components had slight changes in same localities. The oleanolic acid had certain correlativity with latitude and longitude but other components had no correlativity.

Studies on the chemical constituents of Buxus microphylla

2016/1/15 21:07:59

Component extracted from Buxus microphylla with Chloroform was separated and purified through the silica gel column chromatography,from which nine compounds were isolated and identified as:β-sitosterol(I),stigmasterol(II),daucosterol(III),salicylic acid(IV),homovanillic acid(V),5,4'-dihydroxy-3,3',7-trimethoxy-flavonoids(VI),5,4'-dihydroxy-3,3',6,7-tetracarboxylic oxygen-flavonoids(VII),Cleomiscosin A(VIII),3,5-dihydroxy-4',6,7-trimethoxy-flavone-3'-O-β-D-Gluoside(Ⅸ). Compounds IV,V,VI,VII were isolated from the plant for the first time.

Determination of naringin in toothpaste by high performance liquid chromatography

2016/1/15 21:07:59

A high performance liquid chromatographic method was developed for determining the content of Naringin in toothpaste. The chromatographic analysis was carried out on a Hypersil BDS C18 column(250 mm×4.6 mm,5 μm). The mobile phase was water(A)and acetonitrile(B)with gradient elution(0～15 min,10%～100% B). The flow rate was maintained at 1.0 mL/min; The detection wavelength was set at 283 nm and the column temperature was controlled at 40 ℃,The sample injection volume was 20 μL. The results showed that the mass concentration of Naringin with good linearity vs. the peak area within ranges of 14.55～116.40 μg/mL,r=0.9999. The average recovery was 97.56%. The method was proved to be stable,reproducible and precise,It could be used for determination and quality control of naringin in toothpaste.

Change of microtubule cytoskeleton on mitosis in the shoot apex of Saccharum spontaneum

2016/1/15 21:07:43

The microtubule cytoskeleton was investigated by using cryosectioning and indirect immunofluorescence staining in the shoot apex of Saccharum spontaneum. The results indicated that:there were four basic types of microtubule arrays:cortical microtubule,preprophase-band,spindle and phragmoplast microtubule arrays. Cortical microtubule arrays were predominant in the primary thickening meristem of S.spontaneum,while other three arrays were rarely observed. It may be one of the reasons why S.spontaneum has a narrower stem than other cultivars.

Changes of nuclear ultrastructure,nDNA and antioxidant enzymes in leaves of Lemna minor induced by Cd²⁺

2016/1/15 21:07:43

Damage to nuclear DNA and ultrastructure and changes of antioxidant enzymes in leaves of Lemna minor grown in different Cd²⁺ concentration gradient for 6 days were investigated to clarify the phytotoxicity of heavy metals in aquatic plants. The condensation and marginations of chromatin were observed under the electron microscope when the Cd²⁺concentration was from 5-7 mg/L; nucleolus break up when the Cd²⁺ concentration was 10 mg/L. TdT-mediated dUTP nick end labeling(TUNEL)analysis of cells revealed that the nuclear DNA strand breaks could be identified by labeling free 3'-OH termini,which was the typical feature of the cell apoptosis process. And necrotic injury occurred in 20 mg/L Cd²⁺ group. The activities of antioxidant enzymes could be stimulated exposing to low Cd²⁺ concentration to clean up the active oxygen,and the cell apoptosis happened when their activities declined rapidly at higher Cd²⁺ concentrations. The results also indicated that reactive oxygen species(ROS)and antioxidant enzymes were involved in the apoptosis process.

Preliminary study on selection of seedless Siraitia grosvenorii

2016/1/15 21:07:43

Seedless triploid Siraitia grosvenorii,a kind of new germplasm,has been created via polyploid induction,tissue culture and crossing between diploid and tetraploid. It has been tested that the improved breed of seedless S.grosvenorii is quite different from the diploid cultivars which were grown at present. Some of its organs,such as flower and leaf,are lager. It grows more stalwart and blossoms earlier. The fruits have no seeds or very few seeds and 36.28% higher content of glycocides Ⅴ than its diploid female parent. The improvement of biology characteristics of seedless S.grosvenorii,especially the raise of glycocides Ⅴ and the enhance of resistance,is greatly valuable. This paper has reported the growth and development habits,growth stages,form characteristics and fruit quality of the improved breed of seedless S.grosvenorii.

Measurement of coldness by electrical conductivity method in associated with the Logistic Equation in coconut seedlings

2016/1/15 21:07:43

One-year-old leaves of 5 coconut cultivars were used to detect relative conductivity under different cold temperature treatment. The semi-lethal temperature(LT₅₀)was conducted by using conductance method with Logistic Equation. The results showed that the relative conductivity were increased as temperature decreased. The hardiness of the tested cultivars was listed as Hainan local tall coconut>Yellow Dwarf coconut>Red Dwarf coconut>Hybrid of coconut>Aromatic coconut,the semi-lethal temperature ranged from 7.34 ℃ to12.44 ℃. This result could provide the theory basis for cold tolerance breeding of coconut.

Establishment of regeneration system of kiwifruit seedlings

2016/1/15 21:07:43

The full and mature seeds of kiwifruit were germinated to obtain the seedlings,and the regeneration system was established by using the stems,petioles and leaves as explants. The results showed that:the concentration of GA3 which was appropriate to treat the seeds was 2.5 mg/L and the treatment time was 8 h; the medium was more suitable for seed germination; the stems and petioles was easier to dedifferentiation than leaves,however,the average emergence rate of leaves was the highest.

Effects of aqueous extract of four mosses on seed germination and early seedling growth of two Polygonaceae plants

2016/1/15 21:07:43

In this paper,the possible influences of the aqueous extracts of four bryophytes(Sphagnum ovatum,Fissidens cristatus,Hypnum callichroum and Polytrichum commune)on the seed germination and young seedling growth of two vascular plants(Rumex crispus,Polygonum cuspidatum)were investigated. The results showed that the aqueous extracts of these four mosses didn't affect seed germination. However,the crude aqueous extracts of H.callichroum,S.ovatum and F.cristatus inhibited the vigor indices of R.crispus.Furthermore,the crude aqueous extract of F.cristatus significantly enhanced its dry seedling mass,and F.cristatus and S.ovatum significantly inhibited its seedling elongation. For P.cuspidatum,the crude aqueous extracts of four mosses had little effects on its vigor indices and dry mass,but the aqueous extract of F.cristatu had significantly negative influences on its seedling elongation. The possible mechanism that the aqueous extracts of these four bryophytes influencing seed germination and seedling growth of these two vascular plants was not determined. More studies are needed.

Change of diastase activity of Puccinellia tenuiflora cv.Qinghai seeds under sodium carbonate stress

2016/1/15 21:07:43

Changes of diastase activity, soluble carbohydrate content and respiratory intensity of germinating Puccinellia tenuiflora cv.Qinghai seeds under sodium carbonate stress were studied. The results were as follows:diastase activity,soluble carbohydrate content,respiration intensity had a positive correlation with germinating process in salt free condition. When the sodium carbonate concentration was more than 5.00%,changes of diastase activity,soluble carbohydrate content and respiratory intensity of germinating P.tenuiflora cv.Qinghai seeds nearly did not happen,but it had a significant negative correlation with each other. Until the eighth day from being bred,changes of soluble carbohydrate content was basic the same to changes of diastase activity and had a significant negative correlation with each other. But after the eighth day,soluble carbohydrate content begin to decrease and diastase activity continue to raise,it showed that soluble carbohydrate had been used, transferred or composted to new material. Why soluble carbohydrate content decreased under sodium carbonate stress was that diastase activity had been restrained. The decrease of hydrolytic enzyme activity and inhibition of the using of storing material and the inhibition of respiration were the main reasons the P.tenuiflora cv.Qinghai seeds could not germinate under sodium carbonate stress.

Effects of fulvic acid on chlorophyll fluorescence parameters and MDA of sugarcane seedling under water stress

2016/1/15 21:07:43

Two sugarcane cultivars,i.e. ROC22# and YUANLIN6#,were used as the plant materials for the experiment. Normal water supply(CK1)and fulvic acid(FA)of five concentration levels,i.e.0 mg/L(water,CK2),200 mg/L,400 mg/L,800 mg/L and 1 200 mg/L were sprayed on the leaves at initial growth stage(four to five leaves)to study the effects of different FA treatments on chlorophyII fluorescence parameters and MDA in sugarcane seedlings under different degree water stress. The results showed that Fv/Fm,Yield and qP was higher in FA treatments than those of control-2 under moderate water stress and severe water stress. Differences in parameters of varieties and among treatments were found. Comparing with the CK1,the difference of qN among treatments is not obvious. The higher the FA concentration treatments were,the lower the content of MDA was. The content of MDA was significantly lower in FA treatments than that of control-2. On the whole,spraying FA can improve drought resistance at initial growth stage in ROC22# and YUANLIN6# under water stress. The effects of 200 mg/L and 400 mg/L FA concentration treatments were better than those of other FA concentration treatments.

Physiological response of Astragalus fenzelianus and Vicia amoena to low temperature stress

2016/1/15 21:07:43

Effects of low temperature stress on physiological characteristics of A.fenzelianus seedlings and V.amoena seedlings were studied. The results showed that the conductance rate,the soluble sugar content,the proline content and the SOD activity were increased. Taking the conductance rate and the content of soluble sugar as index,the increased degree in A.fenzelianus seedlings was significantly lower than that of V.amoena seedlings at 5 ℃ and significantly higher than that of V.amoena seedlings in -5 ℃ contracted with the 5 ℃; taking the proline content as index,the increased degree in A.fenzelianus seedlings is significantly higher than that of V.amoena seedlings at 5 ℃ and significantly lower than that of V.amoena seedlings in -5 ℃ contracted with the 5 ℃; taking the SOD activity as index,the increased degree of A.fenzelianus seedlings was significantly higher than that of V.amoena seedlings at 5 ℃ and in 5 ℃ contracted with the -5 ℃. Integrating the physiological characteristic indices,it was considered thatA.fenzelianus seedlings showed stronger cold resistance ability to low temperature than V.amoena seedlings. It's therefore concluded that these two kinds of seedings responds differently to low temperature in terms of physiological characteristics and the conductance rate,and the content of soluble sugar is more sensitive to low temperature.

Recombinant aequorin luminous protein and its application in measuring calcium signal of plant cell

2016/1/15 21:07:43

Recently,the application of recombinant aequorin luminous protein has appeared in measuring calcium signal of plant cell as a new method. This paper introduced mainly the development of the recombinant aequorin luminous protein as a probe,the principle and operates method of measuring calcium concentration,and how to conversion the calcium concentration. It summarized the advantages and deficiencies of the protein,and collected mainly the progress in development studies in measuring calcium signal of plant cell. Meanwhile,using the method provided by other laboratory,the authors got the cold shock kinetics of 〖Ca²⁺〗_cyt and 〖Ca²⁺〗_md for the first time.

Role of systemin and jasmonic acid in systemic wounding response of tomato

2016/1/15 21:07:43

Plants produce systemic wounding signals in the wounding site which leads to the expression of systemic defense gene when plants receive the mechanical wounding or insect attack. Among the systemic defense gene,protease inhibitor gene is a representative sample. Tomato is a good model plant in studying the systemic wound response. Three type of wound-response mutants have been identified in tomato plant at present time,through the function analysis of the tomato wound-response mutants and the reciprocal grafting experiment among them,the results indicate that systemin and jasmonic acid activate defence gene systemic expression through a common signaling pathway. Systemin or its precursor protein prosystemin act at or near the site of wounding to increase jasmonate acid synthesis to a level that is required for the systemic response and jasmonic acid or its derivatives act as a mobile wound signal to induce the systemic defense gene expression. The systemic wound signal pathway in plant can be regarded as the inflammatory response in animal,they have lots of similarities.

Abscisic acid catabolism and 8'-hydroxylase, a key enzyme in the oxidative catabolism

2016/1/15 21:07:19

Abscisic acid(ABA)plays great roles in normal growth and development as well as in adaptive responses to environmental stresses in higher plants. For correcting and accurate actions,physiologically active ABA level is controlled through fine-tuning of de novo biosynthesis and catabolism. Hydroxylation at the C-8'position of ABA is the predominant ABA catabolic pathway in higher plants,which is catalyzed by ABA 8'-hydroxylase,a member of cytochrome P450(P450)series. Recently,the CYP707A family of Arabidopsis has been identified as ABA 8'-hydroxylase by genomic and biochemical approaches. The CYP707A family is present in a wide range of plant and functions in ABA catabolism in plants. This paper summarizes ABA catabolic pathways in higher plants and briefly concentrates on 8'-hydroxylation pathway. The key enzyme in the pathway and the genes coding for it are also described.

Effect of exogenous NO on the growth and inorganic ion content of Festuca arundinacea seedlings under Pb²⁺ stress

ZHANG Yuan-Bing, LIU Ai-Rong, ZHANG Xue-Ping, HUANG Shou-Cheng — 2016/1/15 21:07:19

Effects of exogenous nitric oxide(NO)on the growth and some inorganic ions changes of Festuca arundinacea seedlings under Pb²⁺ stress(1 000 mg/L)were investigated. The result showed that:in comparison with the control,Pb²⁺ stress decreased the fresh weight,dry weight,water content,the root activity,inorganic ions contents such as NO^{^</sup>^-₃,K⁺,Na⁺,Ca²⁺,Mg²⁺and Mn²⁺,but increased the root MDA content,the root membrane permeability and Pb²⁺content of F.arundinacea seedlings. In comparison with Pb²⁺ stress,SNP 0.01-1 mmol/L made the fresh weight,dry weight,water content,the root activity,the inorganic ions contents of the seedlings under Pb²⁺ stress increase first and then decline,and the root MDA content,the root membrane permeability and Pb²⁺ content changed in the opposite direction. The results suggested that there was the dose effect of exogenous NO on the seedlings growth under Pb²⁺ stress,suitable concentration of NO could matigate lead poison and enhance the lead tolerance of F.arundinacea seedlings.}

Effect of environmental light intensity on the morphological microstructure and photosynthetic parameters in the cotyledons of Calycanthus chinensis

CHEN Mo-Shun, KE Shi-Sheng — 2016/1/15 21:07:19

The cotyledons of Calycanthus chinesis played critical roles in forming annual seedlings. In order to investigate the mechanism of light adaption of C.chinesis cotyledons,different layer adumbral nets were used in this experiment,the light intensity were 4%,22%,50% and 100% of full sunlight,respectively. The results indicated that with the increasing of shade,the cotyledons showed lower light saturation point(LSP),lower light compensation point(LCP),thinner blade thickness and higher chlorophyll contents. The cotyledons developed fully with the compactness cells and the upgrowth vascular tissues,consequently,the maximal net photosynthetic rate(P_max)and dark respiration rate(R_d)were highest under 50% full sunlight. When the light intensity was below 22% of full sunlight,the cotyledons trended towards the shade-leaved structure. The present study indicated that the cotyledons of Calycanthus chinensis were tolerant to canopy.

Application of proteomics technology on researches of adaptation mechanisms of plants to adverse stresses

2016/1/15 21:07:19

Expression profiling is an important tool to investigate how plant responds to environmental changes. Plants have the ability to alter their gene expression patterns dramatically in response to environmental changes. The proteome profile reflects the actual state of the cell or the tissue of plant and is an essential bridge between the gene transcription and the actual metabolism. In recent years,due to the rapid developments of techniques in protein separation and protein identification technique based on Mass Spectrometry and Bio-informaics,the proteomics technique has become an important technical approach for plant researchers to assess the changes in protein types and their expression levels under different stresses,and many new insights were provided by using this technique. Present paper reviewed the research progresses of plant proteomics on plant responses to abiotic or biotic stresses,including high temperature,low temperature,salt,drought,pathogens and insects.

Variation of seedling physiological biochemical indexes and early selection of Ginkgo biloba

2016/1/15 21:07:19

In this paper,physiological biochemical indexes and growth characters of different Ginkgo biloba family were studied. The main results were as follows:the differences in physiological biochemical indexes and growth indexes were all significant. The correlation analysis showed that correlation coefficient between NR activity and height,seedling biomass both reached positive significant level,correlation coefficient between PPO activity and height,seedling biomass both reached negative significant level. So they could be selected as early selecting indexes.

Morphogenesis of the protocorm of Cattleya hybird in vitro

DING Lan, LI Juan, YANG Hong, QI Lin-Lin — 2016/1/15 21:07:19

Formation and morphogenesis of the protocorms of Cattleya hybrid were investigated in cytology by light microscopy and electronic microscopy. The results indicated that the protocorms consist of numerous parenchymatous cells coming from proliferation of few embryonic cells originated from epidermal cell or subepidermal cell or parenchymatous cell of mother protocorms. While protocorm was cultured on differentiation medium, the meristematic dome was formed at its anterior end,which developed into a bud subsequently. The differentiation of the conducting tissue always occurred at the opposite end. The developmental mode of the protocorms in vitro is very similar to that of its zygotic embryo. A lot of starch grains were accumulated in the chloroplasts of parenchymatous cells when the protocorms grew up and the starch grains were decomposed during the process of morphogenesis.

Tissue culture of Adelostemma gracillimum seedlings and the inducement of lipoxygenase

2016/1/15 21:07:19

Lipoxygenases are important enzymes in plant octadecanoic acid pathway. Products of this enzyme have important functions in plant growth and development,as well as in response to environmental stress. Culture system of Adelostemma gracillimum was the first time to be constructed in Asclepiadaceae. Chitosan was used in the inducement experiment on A.gracillimum seedlings and the products of LOX were detected through LC-ESI-MS. The results showed that A.gracillimum seedlings induced by 150 mg/L chitosan for 9 h produced 9-LOX and the enzyme can catalyze linoleic acid to 9,10,11-trihydroxy-12-octadecenoic acid. It was speculated that octadecanoic acid pathway induced by chitosan differentiated to 9-LOX orientation.

Structure of cell and histology around sprout tumble of Pinellia ternata

2016/1/15 21:07:19

To study the change of Pinellia ternata around sprout tumble under high temperature stress and explore the biological mechanism about sprout tumble, high temperature stress was given when the petiole of Pinellia ternate was 15 cm high. At different days after treatment, the petiole was cut into 3-5 mm, then used for paraffin microtomy. The slices were observed under microscope and pictures were taken. The results were as follows: with the stress time increasing, the fibrovascular tissue structure of petiole was destroyed more seriously, the vascular bundles and cell membrane were broken, the protoplasm spilled over, and the nucleolus got out and even disappeared in some cells.

Vacuolar sequestration mechanisms of anthocyanins in higher plants

2016/1/15 21:07:19

This review sumed up the mechanisms of anthocyanins being sequestered into vacuole in the cells of higher plants. After being synthesized by the multienzyme complexes locating at the cytoplasmic face of the rough endoplasmic reticulum in the cytoplasm,anthocyanins are enwrapped by membrane to form vesicles which migrate mainly toward the vacuole,coalesce each other to form larger vesicles in the migration,and ultimately bring the anthocyanins to the surface of the tonoplast. In most cases,anthocyanins are expeditiously transported into vacuole by various transporters locating on the tonoplast. Other two minor possibilities are that:(1)the vesicles fuse directly with the vacuole;(2)the tonoplast may independently form a large tubular invagination from which the vesicles“bud off”into the vacuole lumen. In the concrete courses of above possibilities,anthocyanins are sequestrated into vacuole in two forms,namely non-modified and modified. The transtonoplast transport of anthocyanins may be accomplished by four models,namely mediated by ATP-binding cassette(ABC)type transporter,by pH-dependent transporter,by the proteins derived from the 24-kD vacuolar protein(VP24)precursor and by multidrug and toxic compound extrusion(MATE)family transporter. It is speculated that different plant species utilize different sequestration mechanisms to accumulate anthocyanins in vacuole,and multiple mechanisms may be simultaneously used in individual plant species.

Studies on factors affecting aucubin production from the suspension cell of Eucommia ulmoides

2016/1/15 21:07:20

Factors affecting aucubin production from the suspended cell of Eucommia ulmoides,such as the different exohormone combination of 2,4-D,NAA,6-BA,KT and nutrients including casein hydrolysate and yeast extract,were investigated. The results showed that the production of aucubin could be improved in media with 2.0 mg/L 2,4-D,1.0 mg/L NAA,0.5 mg/L 6-BA and 0.5 mg/L KT. Nutrients including casein hydrolysate and yeast extract could improve the production of aucubin,the optimal concentration was 100 mg/L casein hydrolysis and 15 mg/L yeast extract. Therefore,the optimal conditions for the production of aucubin were MS media with 2.0 mg/L 2,4-D,1.0 mg/L NAA,0.5 mg/L 6-BA,0.5 mg/L KT,100 mg/L casein hydrolysate and 15 mg/L yeast extract.

Isolation and expression analysis of an OsWNK gene in rice

2016/1/15 21:07:20

BPHiw008,an EST(expressed sequence tag)which is highly homologous with Arabidopsis WNK1(with no lysine kinase),has been screeded from the rice cDNA SSH library. Using this EST as probe to screen the cDNA library induced by brown planthopper sucking,a full length of cDNA with 2.4 Kb in size was isolated and named as OsWNK. OsWNK is deduced to encode a 677-amino-acid protein OsWNK through BLAST. Only 3 amino-acid residues of OsWNK are different from those of AtWNK1. Result of RNA gel blot analysis showed that OsWNK was up-regulated in brown planthopper-sucked rice,indicating that OsWNK may play a role in resistance to brown planthopper of rice.

Estimating copy number of transgenic gene by real-time fluorescent quantitative PCR(TaqMan)

2016/1/15 21:07:20

Real-time fluorescent quantitative PCR is a new technique which has been developed to be used in quantitative analysis of genes as well with the advantages of simpleness,quickness,short needs of target fragment DNA and none of hazardous radioisotopes in recent years. The development,the principle of real-time fluorescent quantitative PCR(TaqMan)and the techinical flow of estimating copy number of transgenic gene are introuduced in this paper.

Esteras isoenzymes analysis of different germplasm materials of Ilex kudingcha

2016/1/15 21:07:02

The difference of esteras isoenzymes of 50 germplasm materials of Ilex kudingcha was compared in this thesis by means of polyacrylamid gel electrophoresis. Jaccard similarity coefficient of 50 germplasm materials was calculated,and the UPGMA was used for cluster analysis. The results showed that there were 13 esteras isozymes bands observed in all test materials,esteras isoenzymes of all test germplasm materials were obviously different,which embodied on the ratio of flow(Rf),the number of esteras isoenzymes bands and strong or weak of the bands,which showed abundant polymorphism for I.kudingcha. The similarity coefficients among the test materials based on esteras isozymes ranged from 0.3000-1.0000; UPGMA cluster result had batter correlation with the origins of germplasm materials,which could distinguish the germplasm materials from different origin and cluster the germplasm materials from the same origin together. The analysis results of esteras isozymes could be considered as one of the significant references for assessing the genetic diversity and the classification of infra-specific taxa of I.kudingcha.

Changes in content of nutritional components and vigor of seed during fruit development of bitter gourd

ZHANG Man, HUANG Ru-Kui, SUN De-Li, LUO Hai-Ling — 2016/1/15 21:07:02

In order to determine the proper harvesting stage for fruit and seed of bitter gourd,the study was conducted to monitor the changes in the content of total soluble protein,total soluble sugar,vitamin C and the seed germinability during fruit development. The results showed that:(1)the fruit enlarged rapidity from 14 to 26 days after anthesis(DAA)in coincide with the increase of total soluble protein,total soluble sugar and vitamin C contents;(2)from 14 to 22 DAA,seeds dry matter accumulated in a fast rate associated with the decline of seed moisture content. Seed showed germinability since 14 DAA,and enhanced thereafter,the highest germination percentage was obtained in 22 to 26 DAA.

Cloning of soybean glycinin G1 promoter and construction of plant expression vector

2016/1/15 21:07:02

A fragment of glycinin G1 promoter was amplified from soybean cultivars Ji nf37 and Jidou 15,respectively. Sequence analysis indicated that both the fragments are 688 bp and had 96.4%-99.6% homologies compared with the registered sequences in GenBank(Sichuan(DQ250808),Nannong87-c38(AY649096)and Dare(X15121)). The promoter fragment from Ji nf37 contains the completely identical elements for seed-specific expression with DQ250808 except only one base difference existing in the Legumin box,which was supposed to have the activity of seed-specific promoter. It was recombined with the γ-TMT gene and plant expression vector pBG1TMT was constructed. The work laid foundation for metabolic engineering to increase α-tocopherol level in oilseed crop seeds.

Effect of soil nitrogen levels on the growth, photosynthetic properties and artemisinin content of Artemisia annua seedling

2016/1/15 21:07:03

Nitrogen is a major determinant of growth rate in plants. In this study,Artemisia annua were cultured under five soil nitrogen levels for about five months. The effects of different nitrogen levels on the growth,biomass allocation,photosynthetic characteristics and artemisinin content of A.annua were examined. The results showed that,with the increase of nitrogen level,the nitrogen content per leaf mass,maximum net photosynthetic rate,light saturation point and the apparent quantum yield of A.annua increased significantly when nitrogen level were 0-0.4 g·kg^-1,and declined whereafter,which indicated that the photosynthetic properties of A.annua could acclimated adequately to nitrogen conditions. High nitrogen content could enhance the photosynthetic capacity of A.annua,and it could utilize higher light intensity. The root mass fraction and root mass/crown fraction of A.annua increased with the increase of nitrogen level,more biomass was invested into root system,nutrient absorbing organ,which could enhance nutrient-capture ability. With the increase of nitrogen level,leaf mass fraction of A.annua increased when nitrogen level were 0.1-0.6 g·kg^-1,more biomass was invested into leaf,assimilative organ,which could increase their carbon accumulation and improve their competitive abilities. Under 0.4 g·kg^-1 nitrogen level,the maximum net photosynthetic rate,diameter,leaf biomass and total biomass of A.annua were maximal,and the artemisinin content was maximal under 0.2 g·kg^-1 nitrogen level. It was suggested to supplement with 0.2 g·kg^-1 nitrogen during the course of cultivation.

Tests on in vitro germination of Betula Luminifera Pollens

2016/1/15 21:07:03

In this paper suitable temperature and optimized medium were determined for in vitro germination of Betula luminifera pollens through comparison tests under different temperatures and on serials of culture media containing various concentrations of sucrose,H₃BO₃ and Ca(NO₃)₂. It was indicated that the germination of B.luminifera pollens could be promoted in the sucrose(basic)media supplementing suitable concentration of H₃BO₃ and Ca(NO₃)₂,while excess of sucrose,boron or calcium could inhibit their germination; the suitable temperature was 30 ℃ and the optimum culture medium was 10% sucrose+200 mg·L^-1 H₃BO₃+200 mg·L^-1 Ca(NO₃)₂ for pollen germination of this species; under these conditions,the pollen germination rate and pollen tube growth became stable after 4 h and 6 h,respectively.

Comparative experimental studies for cold resistance of five plant species

2016/1/15 21:07:03

Five plant species including four kinds from America and one local species were selected to plant materials for comparing the ability to cold resistance using freezing LT₅₀ based on electrical conductivity method and fitting in with Logistic function between percentage of electrolyte efflux from plant tissues and temperatures of treatment. It was shown that kinds of plants' percentage of electrolyte efflux tend to increasing,and take on an S-curve growth. local Savin Juniper was the strongest to resist cold. Its freezing LT₅₀ was -46.58 ℃. The LT₅₀ of both Fourwing Saltbush were -42.44 ℃ and -40.09 ℃ respectively. The cold resistance ability of Prostrate Broomsedge was the lowest. Its LT₅₀ was only -22.18 ℃.

Cellular localization of MAR binding protein of Dunaliella salina

2016/1/15 10:59:06

To study the function of Matrix attachment region(MAR)binding protein(MBP),full length,N-terminal and C-terminal of MBP gene from Dunaliella salina were amplified by RT-PCR,and fused to the green fluorescence protein gene to construct the eukaryotic vector. Then,the recombinant vectors were transfected into Chinese hamster ovary(CHO)cells by Lipofectamine 2000. Fluorescence microscope and western blotting were performed to determine the expression of the fusion proteins and cellular localization. The results showed that the fusion proteins were expressed in CHO cells,the proteins of MBP and C-terminal part were localized in the nucleus and clearly detected in nucleoli,however, the N-terminal part was distributed into all cells.

A practical method for extracting total RNA from flower and fruit of Litchi

2016/1/15 10:59:06

A practical method for extracting high-quality and productive total RNA from flower and fruit of seedless litchi was introduced. These RNA samples thus prepared meet the needs of many molecular biological researches such as constructing cDNA library,RT-PCR,Northern blot and studies on difference analysis of gene expression.

Primary study on main characters of the genetic variation of Winterfat(Ceratoides lanata)

2016/1/15 10:59:06

The initial analysis of 12 main characteristics of genetic variation of Winterfat in four generations showed that 12 main characteristics variation were not significant within the generations,but the number of branches and the number of male flowers,other variations of the 10 traits have reached a significant or quite significant level,showing that the main variations of all characteristics were among generations. The genetic variation coefficient and phenotype variation coefficient had the same overall trend,but there were differences in degree,displaying the potential for genetic variations in these traits is great; the trait herhabilities in plant height,number of females flowers in axillary,leaves width and renewable branches were high a bit(72-62),and the variations were mainly decided by genetic elements,however,the herhabilities of leaves length and plant spread were lower,and its variation was mainly decided by environment factors.

Changes on the edogenous phytohormone content of the Davidia involucrate seed during dormancy and germination process

2016/1/15 10:59:06

Davidia involucrate is an ornamental tree native to China and one of the very rare and endangered plants,which has double dormancy as well as a long dormant period. In the study,dormancy and germination process of Davidia involucrate seeds was divided into four stages according to radicle length. The changes of ABA(Abscisic acid),IAA(Indole-3-acetic acid),GA(Gibberellin)and KT(Kinetin)content in seeds and endocarp were determined by using HPLC among four stages. In addition,the dynamics on GA/ABA,IAA/ABA,KT/ABA and KT/IAA ratio were analysed among four stages. Finally,above-mentioned data were compared with those in seedlings stage. The results showed that:(1)the endogenous hormones content order is ABA>GA>IAA>KT in endocarp of uncracked seeds,the four endogenous hormones content decreased significantly after seeds were cracked;(2)except ABA,GA,IAA and KT contents in seeds increased gradually with radicle elongation. However,the GA,IAA and KT contents were less compared to those in seedlings;(3)in addition,GA/ABA,IAA/ABA and KT/ABA ratio in seed increased gradually with radicle elongation. The change in GA/ABA ratio was greatest than the other two ones. So it is suggested that the trade-off between ABA and GA contents could control the dormancy and germination process in Davidia involucrate seed.

Preliminary study on the trypsin inhibitors in the sweet potato and peanut

2016/1/15 10:59:06

The protease inhibitor inhibiting animal digestion exists in many plant products. Now,trypsin inhibitors have been isolated and purified from the beans and vegetable seeds. In this work,trypsin inhibitors were extracted and purified from peanut and sweet potato by DEAE-Sepharose 4B FF column. Then the inhibition activities of the trypsin inhibitors were determined using the BAEE as substrates. The relative molecular weight(Mr)and isoelectric point(pI)of trypsin inhibitors were determined by the SDS-PAGE and isoelectric focusing electrophoresis respectively. The results showed that there were at least four components of the trypsin inhibitors in the sweet potatoes. Mr was about 20-25 kD,pI was between 5.0-6.6 and there were at least three components of trypsin inhibitors in the peanuts,Mr was about 30-70kD,pI was between 5.0-5.8.

Drought tolerance of indica/javanica recombinant inbred line at germinating stage in rice

2016/1/15 10:59:06

Drought tolerance of indinca/javanica recombinant inbred line(RIL)at germinating stage in rice by use of 20% PEG-6000 as water-stress agent were studied. The radicle length,shoot length and the dry weight of shoot were measured after the 20% PEG treatment for 7 days. The final class of drought tolerance,derived from the sum of the drought tolerance class of each trait,was used as index in assessing the drought tolerance of RIL.The results showed that eight lines are of drought tolerance among the 270 lines,which could be used as drought tolerance resources in rice breeding and research on mechanism of drought tolerance in rice.

Tissue culture of Tripterospermum discoideum

2016/1/15 10:59:06

The tissue culture and rapid proliferation techniques of Tripterospermum discoideum were studied by using stems with buds, stems without buds and leaves as explants. The explants were cultivated in different MS media with different types and concentrations of plant growth regulators. The main results can be concluded as follows:the stems with buds were the best material in speeding propagation among the three explants. The shoot differentiation was MS+BA2.0 mg/L+saccharose3.0%,the optimum medium for proliferation was MS+BA2.0 mg/L+NAA0.1 mg/L+saccharose3.0%,and best medium for rooting was 1/2MS+NAA0.5 mg/L+saccharose1.5%. At the same time,the mode of plant regeneration of T.discoideum in tissue culture has been discussed.

Condition optimization for tissue culture of Arachis duranensis

ZHANG Zhen-Xia, ZHENG Yu-Zhong — 2016/1/15 10:59:06

Condition optimization for tissue culture and regeneration of Arachis duranensis was studied. The results showed that the leaflet was the best experimental explant; the optimized medium for callus was MS+0.5 mg/L6-BA+0.2 mg/L2,4-D or MS+0.5 mg/L6-BA+0.2 mg/LNAA with illumination of 12h/d; the optimized medium for differentiation was MS+1 mg/LTDZ+2 mg/L6-BA+0.5 mg/LNAA; the optimized medium for root induction was 1/2MS+1 mg/LNAA+1 mg/LPP₃₃₃.

Optimization in vitro culture conditions for Gymnema sylvestre

2016/1/15 10:59:06

The optimum medium for tissue culture of Gymnema sylvestre were studied by using germfree tub-seedling as explants. The effects of plant growth regulator(NAA,TDZ,2-ip,IAA,6-BA and KT)on the seedlings proliferation were studied by complete experiment and orthogonal design. The results showed that the optimum basic medium for G.sylvestre was N68; N68+NAA 0.05 mg/L+2-ip 0.4 mg/L+TDZ 0.001 mg/L was the best for bud proliferation,and can be used in subculture; N68+NAA 0.05 mg/L+TDZ 0.005 mg/L promoted rooting,can be used as rooting medium.

Effects of different plant hormones on flower induction of Rhododendron pulchrum

ZHAO Jian, QIU Shuo, LI Xiu-Juan, ZHANG Cui-Ping — 2016/1/15 10:59:06

Effects of GA₃,NAA,IAA on flower bud growth and flowering stage,flowering quality of Rhododendron pulchrum under supplementary light and temperature conditions were studied. The results were as follows:(a)effects of flower induction of Rh.pulchrum appeared to increase initially and then reached their peaks at 2 000 mg·L^-1 and tended to decline later on with GA₃ treatments,and it mainly displayed that the flower bud grow rapidly,the time of flowering stage promoted and the flowering quantity improved. But there were no effects on flower induction of Rh.pulchrum treated with 0-3 000 mg·L^-1 NAA and 0-3 000 mg·L^-1 IAA.(b)flower bud of Rh.pulchrum was significantly positively correlated with the time of flowering stage promoted and flower diamiter,but it was negatively correlated with flower bud stage. And flower bud stage was negatively correlated with the time of flowering stage promoted(P<0.01).

Effects of Glomus versiforme on active oxygen metabolism of Poncirus trifoliate under iron deficiency and heavy bicarbonate stress

2015/12/16 11:38:17

Effects of protective or defensive system on trifoliate inoculated arbuscular mycorrhizal fungus，Glomus versiforme，were investigated on sand culture under iron deficiency and bicarbonate stress. The results showed that the soluble protein contents and the SOD，POD and CAT activities of root or leaf were significantly higher in mycorrhizal plants than that in nonmycorrhizal plants under iron deficiency and bicarbonate stress. Additionally，carotinoid contents in leaves were enhanced and malindialdehyde contents in both leaves and roots were significantly decreased，which suggested that the arbuscular mycorrhizal fungus G.versiforme was able to enhance the defensive ability of plant itself to avoid harm on cells of trifoliate.

Effects of SNP and cPTIO on physiology of Arabidopsis seedling under NaCl stress

2015/12/16 11:38:17

Three groups of 8weekold Arabidopsis seedlings were cultured，they were respectively pretreated with water，0.5 mmol/L SNP and 200 mmol/L cPTIO for 2 h，after that，200 mmol/L NaCl was added. Fresh NaCl solution was applied every day and some physiological indices such as peroxidase(POD)，superoxide dismutase(SOD)，malondialdehyde(MDA)and superoxide anion(O2)were determined 12 h later. The results showed that SNP could relieve NaCl stress injuries，and promote the growth of seedlings under salt stress，significantly improve the antioxidant enzyme system，POD and SOD activity，significantly reduce MDA and O2 content，thereby enhancing the salt tolerance of plants，while the NO specific scavenger cPTIO could reverse the above effects of SNP.

Effects of nitric oxide on seed germination and physiological reaction of maize seedlings under water stress

CHEN YinPing， TAO Ling， YANG Li， WANG Hui， SI XianPei — 2015/12/16 11:38:17

The effects of sodium nitropprusside(SNP)，an exogenous nitric oxide donor at different concentrations，on the seed germination，growth and physiological reaction of maize seedlings under 0.4mol/L manntol(M)stress were studied. The results showed that：(1)The water stress inhibited significantly the maize seed germination and growth of seedlings，increased significantly the malondialdehyde(MDA)content，cell membrane permeability and content of proline；(2)The treatments with SNP at different concentrations could promote the germination rate，germination energy，index of germination and index of vigor of maize seeds，increased the root length，stem length，root height，dry weight，relative water content and carotenoid contents of maize seedlings，inhibited significantly the MDA content，cell membrane permeability and the content of proline of maize seedlings under water stress. It was shown that exogenous nitric oxide had abating effect on depressed seed germination and growth of maize seedlings，could improve the antioxygen ability of maize seedlings，debasing lipid peroxidation and protect plasma membrane from being destroyed during the water stress to enhance the drought resistance of plants under water stress and the best abating effect occurred with the SNP concentrations set at 100 μmol/L and 200 μmol/L.

Analysis on nutritional components and poisonous elements in the leaf of six colorleaved plants

ZHANG Ying， LI XinLei， LI JiYuan， FAN ZhengQi， TIAN Min， CHEN Sheng — 2015/12/16 11:38:17

In this study，nutritional components and poisonous elements were determined in six colorleaved plants，namely P.ceraifera“Pissardii”，L.chinense var.rubrum，P.serrulata，A.palmatum “Atropurpureum”，A.palmatum and L.vicaryi. The results indicated that the maximum protein and soluble sugar content were found in L.chinense var.rubrum，and the maximum vitamin C content was detected in A.palmatum“Atropurpureum”. The contents of βcarotene in the six plants were different obviously，and the highest content was 15.99 mg•kg1，which found in P.ceraifera“Pissardii”. The content of total nutrient elements in L.chinense var.rubrum was highest. There were eighteen amino acids in the six colorleaved plants，and the contents of essential amino acid and total amino acid were all different，but the proportion of essential amino acid in total amino acid had no obvious difference，all were more than 40%. There were certain amount Pb，Cd，As and Hg in the six plants，and the contents of these poisonous elements were all lower than permissible amount，especially As and Hg.

Relation between callus growth speed and somatic embryogenesis of citrus

ZHANG JunE — 2015/12/16 11:38:17

In order to discuss the cause of loss of regeneration capability of citrus callus，and to look for the relation between callus growth speed and somatic embryogenesis，in this study，calli growth speed of twentynine genes from seven citrus types were detected，and the correlation between callus growth speed and somatic embryogenesis was statistically analyzed. The results showed that correlation coefficient of the growth speed and somatic embryogenesis was 0.3683. It was thus deduced that other influence factors still occured between them.

Study on regeneration system from callus of Rehmannia chingii falbiflora

MA DanDan， XIA GuoHua， JIN FeiFei， LI GenYou — 2015/12/16 11:38:17

Rehmannia chingii f.albiflora is a particular and scarce forma of R.chingii. In the present research，its young leaves were used as explants to study the effects of the growth regulators on callus induction and plant regeneration. The results indicated that the optimal media for induction and differentiation of callus were MS+BA1.5 mg•L^-1+IBA0.5 mg•L^-1 and MS+BA2.0 mg•L^-1+NAA0.1 mg•L^-1，respectively；the effective medium for cultivating adventitious buds was MS+BA2.0 mg•L^-1+IBA0.2 mg•L^-1; moreover，it’s found that the best inducing combination for root formation was 1/2MS+NAA0.05 mg•L^-1；the survival rate of testtube plantlets could reach 96.7 % after transplanting. In addition，the prospects of its gardening application and medicinal value were discussed.

Characteristics of potassium channel of different tobacco genotypes

LU LiMing — 2015/12/16 11:38:04

The characteristics of root cortex protoplast whole cell transmembrane inward K+ current of four fluecured tobacco genotypes with different potassium use efficiency were studied. The results showed that the current density (pA/pF) of NC89 （80） was the highest of all four genotypes，then Nongda 202，Jingyehuang and NC2326 were 60，50 and 40 respectively. The results suggested that the current density (pA/pF) of transmembrane inward K+ current was closely related to the potassium absorption ability and potassium use efficiency of fluecured tobacco，and could be used as a effective method to screen high potassium use efficiency fluecured tobacco genotype.

Seedless fruits producing with somatic hybrid as male parents

2015/12/16 11:38:04

Diploid Shatian pummelo was used as the female and crossed with the allotetraploid somatic hybrids(Citrus jambhiri+C.sinensis(HR)；C.grandis+Rough lemon(SR)；C.aurantifolia+C.sinensis(KV)to generate seedless Shatian pummelo，at the same time，the fruits’qualities were analyzed(using Citrus grandis pollen as the control). Results showed that the seed abortion phenomena were very clear. The rates of deflated fruit seeds of treatments were 41.1%-96.0%，the corresponding value of control was only 2%. There were no significant differences between treatments and control including average fruit weight，pulp weight，pericarp weight，pericarp thickness，fruit shape，percentage of edible and solubility of solid substance，as well as total sugar，Vc and titratable acid content，though some values were lower than those of control. Two suitable allotetraploid somatic hybrids(HR and SR)for seedless fruits producing were selected(the rates of seedless seeds were over 89.1%).

Differentially expressed gene of indica/javanica recombinant inbred line in rice at early stage of PEG stress

2015/12/16 11:38:04

The drought tolerant line and drought sensitive line screened from the population of indica/javanica recombinant inbred line were stressed with 20% PEG(Polyethyleneglycol) for 6 h at fiveleaf stage of rice. Ten of cDNA fragments were isolated by mRNA differentially display reverse transcription PCR(DDRTPCR)，but only four fragments were approved truth by reverse Northern dot blotting. These fragments named as DTDF1，DTDF2，DTDF3 and DTDF4 were sequenced and then analyzed by Blast program. The putative protein of DTDF3 was highly homologous with NBSLRRlike protein B and that of DTDF2 was homologous with Llactate oxidase. The putative protein of DTDF1 was not found homologous protein，and that of DTDF4 was highly homologous with a hypothetical protein.

Effect of NO-3N concentration on nitrate nitrogen metabolism of winter wheat seedlings under water culture

2015/12/16 11:38:04

In cultural liquid Hoagland，with winter wheat set as testing object，this research tested dynamically the absorption and accumulation of nitrate nitrogen，the activities of nitrate reductase，the content of ammonium nitrogen，the nitrogen absorption of the wheat and root activity under the three levels of nitrogen contenthigh，medium and low. This research completely analyzed the influence over the absorption，reduction and transferability of nitrate nitrogen of winter wheat being treated with different nitrogen leves，and investigates the change of nitrate nitrogen metabolism on aboveground and underground parts of plant，under different nitrogen levels. The research indicated that，in water culture，the consumption of nutrient solution NO-3，the change of pH values，complete nitrogen of plant and root activity reflected the effect on metabolism of plant nitrate nitrogen well under different nitrogen levels；Under high nitrogen level，assimilation of NO-3in plant was weaker than that of medium nitrogen level as winter wheat accumulates adequate NO-3itself rather than absorbing more from nutritional liquid. Under different nitrogen levels，no relationship between NO-3 supply and plant NRA was found，the curve for the roots and aboveground prat varied，under high NO-3 level，the aboveground part of the plant was the main assimilated part；on the contrary，under low NO-3 level，the root part of the plant was the main assimilated part. Even NO-3 was a safe nitrogen resource，it could restrain the further assimilation of nitrate nitrogen in plants under too much higher level，on the other side，it could lead to an insufficiency absorption of NO-3 and the decrease of root activity，which was not good to the absorption of nitrogen nutrition of young wheat seedling.

Effects of NAA and PP333 on physiological and biological changes of Jasminum sambac

2015/12/16 11:38:04

Different concentrations of NAA(20，50，100 mg/L) and PP333(100，300，500 mg/L) were sprayed on the stem and leaves of Jasminum sambac at early florescence stage，the effects on J.sambac flower buds formation were studied. The changes of soluble protein content，free amino acid content and protective enzymes activities in new shoots after treatment were observed. The results showed that new shoots excessively grew，florescence and growth phase were delayed after NAA treatment. While it was contrary that florescence was ahead of 4 d after PP333 treatment，the yield of flower increased remarkably to 12.95% than those of control, and the optimum concentration was 300 mg/L. The changes of soluble protein and free amino acid content were stable during flower bud growth stage after PP333 treatment. But it was on the contrary after NAA treatment. The POD activity decreased after NAA and PP333 treatments，and higher than those of control after NAA treatment. SOD activity increased after NAA and PP333 treatment. There were different effects after NAA and PP333 treatment，and suitable concentration of PP333 treatment could increase the yield of bud of J.sambac in this paper.

Seed germination and the protocorm multiplication of Anoectochilus roxburghii

2015/12/16 11:38:04

Seed germination of Anoectochilus roxburghii in asymbiotie culture and the effects of four factors(culture medium，6BA，ZT and NAA)on protocorm multiplication through orthogonal design were studied. The results showed that pollination type was an important factor on A.roxburghii seeds' nonsymbiotic germination，germination ratio of the xenogamy seeds reached 78.53%，and the germination ratio of the geitonogamy seeds reached 69.62%，while the germination ratio of selfpollination was only 39.87%. The most suitable maturity of capsule appeared when it grew for 150 days，the germination rate of such seeds could reach 78.59%. The vitality of the seeds were affected by refrigerated storage:the germination rate of the seeds was decreased with the increase of seed cool storage time. There were no significant changes of seed germination rate between seed treatment by 10% NaClO solution and the control. NAA had a significant effect on the proliferation of protocorm，the optimum culture medium for protocorm proliferation was 1/2MS+ZT0.5mg/L+NAA1.0mg/L.

Reveiw on Rehmannia glutinosa breeding

ZHOU YanQing， YAO HuanLing， ZHOU ChunE， DUAN HongYing， LU ShuXia — 2015/12/16 11:37:44

Medicinal plant Rehmannia glutinosa（Scrophulariaceae）is one of “the top four Huai herbal medicines” in China，which is possessed of important medicinical and economical values. This review summarized the achievements made so far in the breeding of R.glutinosa by introduction，selection，hybridization，tissue culture，gene transformation and DNA markerassisted breeding in order to provide reference for the breeding researchers of R.glutinosa.

Category， functions and chromosomal locations of the gene loci related to colored potato pigments

ZHAO ChangLing， LI SunWen*， ZHANG WeiMing — 2015/12/16 11:37:44

This paper summarized the idea origin，category，functions and chromosomal locations of the gene loci involved in the production and distribution of the pigments in colored potatoes. The idea of the gene loci related to the pigments in colored potatoes originated from the two genetic models which attempted to explain the inheritant behaviors of the colorations of the stem tubers and other parts of tetraploid and diploid potatoes. The overall thirteen gene loci related to the pigments in colored potatoes could be classified into four kinds. The first，second and third kinds were related to the synthesis，acylation and distribution of the potato anthocyanins respectively，and the fourth kind was related to the production of the potato carotenoids. Locus I，P，R and Y encoded a MYBdomain transcription factor，flavonoid 3′，5′hydroxylase，dihydroflavonol 4reductase and βcarotene hydroxylase respectively. Various and complicated interactions among the gene loci synthetically determined the production and distribution of the pigments，especially the anthocyanins，in colored potatoes. Locus D and R were located on Chromosome 2 of potato，E，F，I and PSC on Chromosome 10，P on Chromosome 11，and Y on Chromosome 3. This paper could provide a reference for the exploration on the genetic mechanism of the colorations of colored potatoes.

Cell division frequency difference in various regions of stem tip protomeristem in sugarcane varieties with different stem diameters

2015/12/16 11:37:44

In order to understand the correlation between difference of cell division frequency in various regions of stem tip protomeristem and stem diameter as well as plant height，continuous paraffin sections of six sugarcane varieties were investigated by microscopy at five different growth stages. The results showed that there was significant difference in cell division frequency of various regions of stem tip protomeristem. The difference followed the pattern:peripheral meristem(3.89%)＞corpusprimary cell(2.67%)＞pith(1.46%)＞tunicaprimary(1.3%). There was positive correlation between cell division frequency in different regions of stem tip protomeristem and sugarcane stem diameter，and the correlation coefficients were highly significant between stem diameter and pith zone and tunicaprimary cell zone(r2=0.856*and r2=0.925*，respectively). However，there were negative correlations between stem length and cell division frequency of stem tip protomeristem in different zones，and the correlation coefficient between corpusprimary cell and stem length was significant. The results could reveal the inherent relation between different stem tip protomeristem zones and their characteristics with precise quantifications of cell division frequency in different protomeristem zones of sugarcane. The difference of cell activity in different zones could be the basic cytology of sugarcane stem thickening.

Effect of perennial cultivation on “Dong A” genic male sterile lines in annual upland cotton

ZHANG Xin1， CHEN GuoPing1，2， ZHOU RuiYang1* — 2015/12/16 11:37:44

Genic male sterile (GMS) lines of plant has the characteristic which is easy to restore but hard to maintain. On the basis of frostless climate characteristics in winter of Nanning，Guangxi,“Dong A” GMS lines in annual upland cotton was field cultured for three years. The results showed that，biennial and triennial “Dong A” cotton could flower in early May and open bolls in late June，which were hard for annual “Dong A” in the same environment；comparing to annual “Dong A” cotton，the 100seed weight index，bolls per plant，and yield of biennial and triennial “Dong A” cotton were significantly increased，but the boll weight and the lint rate of them were significantly decreased，and the dead locule rate of them was significantly increased； the fiber quality of biennial and triennial “Dong A” cotton was basically not changed significantly. It was concluded that，the utilization of perennial cultivation on GMS lines for stock breeding had a good prospect in the south subtropical zone.

Amylase and formate dehydrogenase isozymes in the genus Fagopyrum

ZHANG YiZhong1，2， CHEN QingFu1* — 2015/12/16 11:37:44

The amylase isozyme and formate dehydrogenase isozyme of 42 accessions of cultivated and wild buckwheat belonging to eight species of the genus Fagopyrum including seven species of the bigachene group and one of the smallachene group were studied by means of polyacrylamide gel electrophoresis(PAGE). The isozyme samples were obtained from dry and sprouting seeds，respectively. The results showed that dry seeds had no amylase activity but the sprouting seeds had very active amylase isozymes. A total of 23 bands for the amylase isozyme were observed in sprouting seeds of cultivated and wild buckwheat，with F.esculentum and F.tataricum having 10 and 8 bands，respectively. Variations in the amylase isozyme zymographs were large among different buckwheat species but were less so within the same species. The results of zymograph analysis and clustering showed that F.megaspartanium and F.pilus are closely related to F.esculentum and F.tataricum，respectively，supporting Chen’s hypothesis that F.megaspartanium and F.pilus are ancestors of these two species，respectively. Only one band was observed for the formate dehydrogenase isozyme in the dry and spouting seeds of all accessions.

Heatshockinduced heat tolerance in maize seedlings and involvement of proline

LI ZhongGuang*， GUO Ying， YANG ShuangMei， ZHANG LuQi， YU Yan， ZHAO YiXia — 2015/12/16 11:37:44

Effect of heatshock(HS)on heat tolerance in maize seedlings and its relation to proline(Pro)were studied in this paper. The results showed that HS pretreatment at 42 ℃ for 4 h followed by a 4h recovery at 26.5 ℃significantly increased survival percentage of maize seedlings under high temperature at 48 ℃，and the procedure of HS and recovery displayed Pro accumulation in maize seedlings. In addition，pretreatment of different concentration exogenous Pro markedly enhanced the level of endogenous Pro and the activities of antioxidant enzyme ascorbic acid peroxidase(APX)，catalase(CAT)，superoxide dismutase(SOD)，glutathione reductase(GR)and peroxidase(POD)in maize seedlings，and improved survival percentage of maize seedlings under high temperature. These results suggested that the increase of antioxidant enzyme activities by HSinduced Pro accumulation inducement could be one of the physiological basis in the formation of HSinduced heat tolerance in maize seedlings.

Tissue culture and plant regeneration in Begonia fimbristipula

CHEN Gang， CHEN XiongWei， WANG YingHua — 2015/12/16 11:37:44

An efficient and rapid regeneration protocol was developed using leaves from geminating seedlings of Begonia fimbristipula. MS medium containing 1.00 mg/L 2，4D，1.0 mg/L 6BA，200 mg/L LH，200 mg/L CH and YE 200 mg/L was most effective in inducing calli. The frequency of callus induction was above 96.88%. Optimal medium for inducing adventitious buds is MS+1.0 mg/L 6BA+200 mg/L LH+200 mg/L CH+YE 200 mg/L，and the highest frequency reached 77.3%. The proper culture medium of rooting was 1/2MS and the rooting rate was 100%. The survival rate of the plantlet was up to 90% after being transplanted into soil.

Effects of the culture conditions on galanthamine biosynthesis in Lycoris aurea calluses

RU QiaoMei1， PEI ZhenMing1，2， ZHENG HaiLei1* — 2015/12/16 11:37:44

The biosynthesis of galanthamine in callus culture of Lycoris aurea was studied. Some factors affected the biosynthesis of galanthamine were investigated. The results showed that sucrose was the best carbon source，and the concentration of 90 g/L was the most preferable for galanthamine production，which was reached to 0.068 %. Meanwhile，adding 500 mg/L Lphenylalanine to medium，galanthamine content was also improved. Tyrosine and salicylic acid were not suitable for galanthamine accumulation in calluses. Besides，2，4D inhibited the biosynthesis of galanthamine. Moreover，NAA and IBA above 0.5 mg/L could restrain the production of galanthamine，but 6BA treated with high concentration promoted the biosynthesis of galanthamine.

Tissue culture and rapid propagation of dwarflate pomelo

2015/12/16 11:37:44

Aseptic seeding of dwarflate pomelo were used as the materials in the study，We induced cespitose buds from meristem and epicotyl of the seedlings，then regenerated plants from the induced shoots，and results showed that both mature and immature seeds could be germinate on 1/2MS and MS medium，the highest germination rate could reach 96%，and the aseptic seeding from mature seeds were more suitable for the further differentiation. The epicotyl was found to be the optimal explants，the optimal protocol for cespitose buds proliferation was to culutre upicotyl on MS medium 〖JP3〗with 2.0 mg•L1 6BA，0.1 mg•L1 NAA and 40 g•L1 sucrose，and the achieved multiplication coefficient was 8.4. The optimal rooting medium was 1/2MS medium with 0.2 mg•L1 NAA，0.2 mg•L1 IBA 0.2 g•L1 AC，〖JP2〗from which 90% rooting rate was obtained. After ransplanted to nutrition bowl，80% plants survived.

Establishment for induction system of female flowers in steriled seedlings of Cucumis sativus

2015/12/16 11:37:28

The synergistic effect of Spm and IAA,and the effects of agar concentration,KH2PO4 concentration on the induction of female flowers in steriled seedlings of Cucumis sativus for the establishment of an induction system of female flowers were reported. It was found that the induction rate and the amount of female flower in every trial were considerably low and even down to zero when Spm or IAA was respectively added to derooted seedlings cultured on MS basal medium,while the induction effect were obviously higher when 12 mg•L-1 Spm and 0.01 mg•L-1 IAA were added simultaneously,and no female flowers could be observed on the control medium,which showed that Spm and IAA had an significant synergistic effect on the induction of female flowers. Experiment was done with different explant of whole seedlings,derooted seedlings,detoped seedlings without root,top shoots or with agar concentration of 0.5%-0.9%,and with KH2PO4 concentration of 1.0-2.0 mmol•L-1,the highest induction effect for female flowers was achieved on medium of agar 0.7%,KH-2PO-4 1.75mmol•L-1 and from detoped seedlings without root,its induction rate and amount of female flower reched to 46%,54 respectively.

Regenerating triploid citrus plants by embryo rescue technique

2015/12/16 11:37:28

Diploid Shatian pummelo was used as the female and crossed with the allotetraploid somatic hybrids(Citrus jambhiri+C.sinensis(HR);C.grandis+Rough lemon(SR);C.aurantifolia+C.sinensis(KV))to regenerate triploid citrus plants by embryo rescue technique,using C. grandis pollen as the control. Results showed that the regenerating rate of treated sexual hybridization was significantly lower than that of the control(P＜0.01),in which the corresponding value of Shatian pummelo×HR was comparably higher(the rate of seedling is 101%). The optimal medium for embryo rescue was solid MT medium supplemented with GA3 1.0 mg/L,queen bee oar 200 mg/L,lacialbumin hydrolysate 250 mg/L and agar 0.7%. Results of flow cytometry analysis showed that triploid citrus plants were successfully obtained in this study.

Influence on microtubule in wheat mesophyll cell exposed to enhanced ultravioletB radiation and He-Ne laser irradiation

GUOAi-Hua,GAOLi-Mei,LIYong-Feng,HANRong* — 2015/12/16 11:37:28

In this paper,the protoplasts from fresh leaves of wheat seedlings was acted as material,which was exposed to enhanced UV-B radiation(10.08 kJ•m-2•d-1),HeNe laser irradiation(5 mW•mm-2 )and the combined treatments of UVB and HeNe laser. Effects of different treatments on microtubule(MT)cytoskeleton,which was labeled with indirect immunofluorescence localization,was studied through confocal laser scanning microscope(CLSM). The results showed that microtubule of protoplast in wheat seedlings was depolymerized significantly to stick and spot under the condition of enhanced UV-B radiation,the MT bundles was trend to dispersivity and decreasing of fluorescence intensity. However,the depolymerization of MT was decreased by He-Ne laser irradiation after enhanced UV-B radiation. Therefore,the damage of microtubule in wheat seedlings induced by enhanced UV-B radiation that can be repaired partly by He-Ne laser irradiation.

Establishment of SRAP amplification system for Rehmannia glutinosa and primer screening

ZHOUChun-E,GUFeng-Ping,LUShu-Xia,DUANHong-Ying,ZHOUYan-Qing* — 2015/12/16 11:37:28

The single factor design was used to optimize SRAP amplification system of 22 Rehmannia glutinosa lines in five factors(the concentration of template DNA,Taq DNA polymerase,primer,Mg2+and dNTP). SRAP amplification system was established as follows:2.5 mmol/L Mg2+,0.30 mmol/L dNTP mixture,2.5 U Taq DNA polymerase,0.32 μmol／L each primer,20 ng template DNA and 2.5 μL 10 X PCR buffer in 25 μL SRAP reaction system were the best suitable PCR system. 12 primers were collected from 88 primers using the optimized amplification system above.

Construction of plant expression vector harboring AtCKX1 Driven by droughtinducible promoter of RD29B from Arabidopsis

GAOXi-Le,WUGuo-Jiang,LIMe-Ru* — 2015/12/16 11:37:28

In this study,the cloning of AtCKX1 and 5′flanking promoter sequence of droughtinducible RD29B from Arabidopsis was reported. The 1705 bp promoter sequence was analyzed bioinformatically in the database of plant cisacting regulatory element (PlantCARE). The result showed that there were several important cisacting elements,including ABRE (ABA response element),Myb binding site,TATAbox,CAATbox,in the 1705bp promoter region. The binary vector harboring AtCKX1 driven by RD29B promoter was further constructed to be used in coming plant transgene.

Effects of salt stress on some physiological characteristics of Spartina anglica seedling

2015/12/16 11:37:28

Under different salinity conditions（0,20,30,50,100 mmol/L NaCl）,variations of some physiological indicators including MDA,free praline,soluble sugar,soluble protein contents and antioxidative enzymes(SOD,POD,CAT)activity of Spartina anglica seedlings were investigated. The results were showed as follows:the antioxidative enzymes(SOD,POD,CAT)activity was gradually increasing during the forty days under salt stress and there were significant differences with the prolongation of time salt stress. Compared with ckeck,the MDA contents generally decreased with the prolongation of time salt stress. The free proline,soluble sugar and soluble protein contents in leaves showed an increasing trend with increasing salinity and the prolongation of time salt stress. Accumulation of osmoprotectants was the key response of S.anglica salinity stress,and the antioxidative enzymes played important roles in it.

Tissue culture and plant regeneration of Sophora alopecuroides

CAO You-Long, LI Xiao-Ying, LUO Qing, BEI Zhan-Lin — 2016/1/15 3:05:17

In order to study the factors that influence the tissue culture of Sophora alopecuroikes,four callus-inducing media,i.e.P1(MS+2,4-D 0.5 mg/L),P2(MS+6-BA 0.1 mg/L+NAA 0.5 mg/L),P3(MS+6-BA 0.5 mg/L+NAA 0.5 mg/L),P4(MS+NAA 1.0 mg/L+KT 0.5 mg/L),three different media(MS+6-BA 1 mg/L; MS+6-BA 0.5 mg/L; MS+6-BA 1 mg/L+NAA 0.2 mg/L),and four root-inducing media(MS; MS+IBA 1 mg/L; MS+IBA 1 mg/L+IAA 0.5 mg/L; 1/2 MS+IAA 0.2 mg/L)were applied. The results showed that the induction frequency of calluses highly depended on the varieties and concentration of hormone in media. 2,4-D in media,with concentration ranging from 0.2-2 mg/L,was favorable to the growth of calluses of S.alopecuroikes,accompanying earlier browning occurrence. It was found that adding active charcoal into medium could efficiently inhibit the callus browning. In addition,IAA seemed an essential ingredient in medium to the root differentiation of S.alopecuroikes.

The work was conducted at the USDA-ARS,Sugarcane Research Laboratory,Houma,LA,U.S.A. under a Non-funded Cooperative Agreement (USDA Control No. 410334)between the Sugarcane Research Center,Chinese Academy of Agricultural Sciences,Nanning 530007 and the USDA- ARS,Sugarcane Research Laboratory,Houma,LA 70360,U.S.AAssessment of genetic diversity in Saccharum using SSR markers and capillary electrophoresis

2016/1/15 3:05:17

This study was conducted to assess the genetic diversity amongst 12 Saccharum clones from 3 species using 19 SSR markers and capillary electrophoresis(CE). A total of 229 DNA bands were generated showing a size range between 100 and 260 bp. Based on the SSR data,the Jaccard's genetic similarity coefficients ranged from a minimum of 0.09 between CP72-1210(cultivar)and In81-142(S.spontaneum)to a maximum 0.65 between GX87(S.spontaneum)to GX86(S.spontaneum). In the dendrogram using UPGMA method,the 12 Saccharum clones were clustered into two groups. The three S.spontaneum clones formed a single group,while the three S.officinarum clones clustered with the six cultivars,which demonstrated that S.officinarum had a closer genetic relationship with cultivars than S.spontaneum. The SSR markers data generated based on capillary electrophoresis are more accurate,simpler and automatic as compared to other molecular markers or electrophoresis.

Establishment and optimization of ISSR-PCR reaction conditions for Rheum tanguticum

2016/1/15 3:05:17

Inter-Simple Sequence Repeats(ISSR)is a good molecular marker for revealing genetic diversity. Reaction system differed in different species,so optimization of ISSR-PCR reaction is very important. Factors which affect the ISSR-PCR amplification,such as the concentration of Mg²⁺,Taq DNA polymerase,dNTP,primer and template DNA with different annealing temperatures,were optimized and selected by using the genomic DNA of Rheum tanguticum as material. Optimal PCR(20 μL)mix contained 1.5×PCR buffer(15 mmol/L Tris-HCl,75 mmol/L KCl),1.00 mmol/L MgCl₂,0.6 U Taq DNA polymerase,0.125 mmol/L dNTP,0.5 μmol/L primer and 30ng template DNA. The suitable annealing temperature was 57.4 ℃ for primer UBC888. Establishment of the PCR reaction conditions could favor further studies on the genetic diversity of R.tanguticum by using ISSR molecular marker techniques.

Some physiological and biochemical charateristics of different developmental stages of somatic embryogenesis in Anthurium andraeanum

2016/1/15 3:05:17

Using Anthurium andraeanum cv.‘Amigo'as material,some physiological and biochemical characteristics of different developmental stages of somatic embryogenesis were studied. The results showed that the activity of POD and CAT maintained at a high level at the stage of embryogenic callus,and SOD activity present a high level at the later stage of somatic embryogenesis. The content of soluble protein was the highest at the stage of callus formation,and the content of starch went down first and up later but the content of soluble sugar up first and down later contrarily during the somatic embryogenesis. SDS-PAGE analysis showed that there are some specific proteins and high expression proteins at the stage of embryogenic callus. It concluded that the embryogenic callus formation stage is important for the regulation of somatic embryogenesis in A.andraeanum.

Advances in biological functions and biosynthesis regulation of tocotrienols

2016/1/15 3:05:18

The molecular and the biosynthetic pathway of tocotrienol were briefly introduced. And the powerful neuroprotective,anti-cancer,cholesterol lowering and antioxidation properties of tocotrienols as well as the strategies of metabolic engineering used in elevating biosynthesis level of the tocotrienols in plants were mainly reviwed.

Effect of exogenous ALA、SNP and Spd on seed germination characteristics of Platycodon grandiflorum under NaCl stress

DU DanDan， HE Ping*， ZHANG ChunPing， YU ZeLi — 2015/12/16 11:39:04

In order to get the method to improve the salt resistance of seeds and seedlings for Platycodon grandiflorum under NaCl stress，the different concentration of 5aminolevulinic acid（ALA），sodium nitroprusside（SNP），spermidine（Spd）effect on P.grandiflorum seed germination，seed germination rate，seed germination index，and the average length of root were studied under different concentrations of NaCl stress. The results showed that the seed germination was inhibited under 75 mmol•L1 concentrations of NaCl stress，but with different concentrations of ALA，SNP and Spd on the P. grandiflorum seeds，all germination index were significantly higher. Spd(2.5 mmol•L1），ALA （5 mg•L1）and SNP（0.1 mmol•L1）could obviously alleviate the damage of salt stress to the seeds of P. grandiflorum. Spd(2.5 mmol•L1)significantly increased all indexes. The germination vigor was 40.3%，the germination rate was 49.5%，the germination index was 9.28 and the average length of root was 2.80 cm. So we conclude that，ALA，SNP and Spd with appropriate concentration could significantly alleviate the damages to the P.grandiflorum seeds under NaCl stress and promote the salt resistance of the seeds，the best of them was Spd，the second was SNP，furthermore was the ALA.

Sequence analysis and function predication of poroxidase from Conocephalum conicum

2015/12/16 11:39:05

Peroxidases is widespread in biological world. It plays an important role in the cell redox metabolism.In order to furher study the characteristics and functions of Conocephalum conicum poroxidase，we performed bioinformatics analysis of its homology，subellular location，tertiary structure，function and so on. The results showed that the sequence had a complete open read frame of about 1 050 bp and encodes 349 amino acids，and the sequence had similar POD with that of Arabidopsis thaliana(the similarity beween them is 71%). This sequence was predicated to be C.conicum poroxidase which belonged to the poroxidase super family，and R68、F71、H72、P170、H200 and D276 to be the C.conicum POD’s active sites in which the four groups of disulfide bonds：C4 and C122、C74 and C79、C128 and C328、C207 and C239 played a very important role in maintaining the protein’s function，natural conformation and stability. This study would set an important foundation for the study of the interaction mechanisms and functions of the C.conicum poroxidase.

Effects of artificial pollination on fruit development of Pitaya （Hylocereus polyrhizus）

2015/12/16 11:39:05

To study effects of pollination on fruit development of Pitaya（Hylocereus undatus），artificial pollination and natural pollination were treated to flowers as well as ovary and fruit in different development phases were dissected and sliced for observation. Results suggested that artificial pollination treatment made average single fruit weight and marketable fruit percentage raised while average single fruit weight was positively correlated with seed quantity in the fruit. Ovary of Pitaya were comprised of numbers of‘PodospermOvule’structure inside. Ovules grew into seeds after flowering while podosperm into succulent seed stalk. Main edible part of fruit were comprised of numbers of‘succulent seed stalkseed’structure. Artificial pollination might increase the numbers of‘PodospermOvule’developing into‘succulent seed stalkseed’that gave help to fruit enlargement against natural pollination group.

Study on the correlation between physiological indexes as ABA， etc and desiccation tolerance in developing chestnut seeds

ZONG Mei1，2， CAI YongPing1， FAN ZhiQiang2 — 2015/12/16 11:39:05

The partial physiological indexes of embryonic axes and cotyledons in the developing chestnut seeds，and the correlation between them and the desiccation tolerance were studied. Experimental results showed that with the development of chestnut seeds，the contents of ABA，starch and soluble proteins of embryonic axes and cotyledons on the 80 days after anthesis reached synchronously the maximum，the content of soluble sugars of embryonic axes and cotyledons on the 80 days after anthesis was the minimum. At the moment the desiccation tolerance of chestnut seeds on the 80 days after anthesis was the strongest，so it should be the optimal harvest period for chestnut. In addition，by correlation analysis，the acquisition of desiccation tolerance in the developing chestnut seed was related to the changes of matter in embryonic axes. It was significantly correlative with starch and the contents of ABA and soluble proteins，and was negatively correlative with the contents of soluble sugars. Therefore，the period of maximum desiccation tolerance in chestnut seeds could be inferred by the change of starch.

Effect of flooding on some physiological indexes of Panicum repens

ZHAN JiaHong， LAN ZongHui — 2015/12/16 11:39:05

Effects of flooding stress on the contents of malondialdehyde(MDA)，soluble sugars and proteins from young Panicum repens，and the activities of three protective enzymes in leaves were studied. The results showed that distinct changes of their contents were induced by the flooding conditions:the contents of MDA slowly ascended；while the soluble sugars first increased and then decreased；and the soluble protein descended gradually. We also observed that the activities of superoxide dismutase(SOD)and catalase(CAT)fell after rising，but the activity of peroxidase(POD)always went up. Together，these results suggested that flooding had an impact on physiological indexes of P.repens，and the plant could effectively adapt to the flooding environment by rising the activities of protective enzymes.

Primary study on suspension cell culture of Epimediun wushanense

HAN SuJu1， LI YunXiang2* — 2015/12/16 11:39:05

The technological system of cell suspension culture by the flavonoids content of E.wushanense was established through studing the effect of inoculation concentration，hormone combination，sugar concentration and various medias. The result showed that the suspension culture optimum medium of E. wushanense was B5 medium supplemented with 1.0 mg•L12，4D and 0.2 mg•L16BA，the sugar concentration was 4.0% and the optimum inoculation concentration was 2 g per 30 mL. The highest content of the total flavonoids appeared on the medium.

Physiological changes and heat tolerance comparison of five Rhododendron species under hightemperature stress

2015/12/16 11:38:50

High temperature is the important constraining factor of Rhododendron in ex situ conservation and landscape application. In order to reveal the injury of mechanism of five Rhododendron species under hightemperature stress，R.mucronatum，R.molle，R.moulmainense，R.chihsinianum and R.rubiginosum which belong to different subg.Rhododendron were used in the experiment. Four yearsold seedlings of these five species were planted in artificial climate chambers and their changes of physiological and biochemical index were determined under hightemperature of 30 ℃ and 38 ℃ stress，and the heat tolerance was comprehensively evaluated with fuzzy subordinate function and system cluster analysis. The results showed that the activities of catalase(CAT)，ascorbate peroxidase(APX)in leaves of five species were increased with temperature becoming higher，so do the contents of proline(Pro)，hydrogen peroxide (H2O2) and malondialdeyde(MDA). The activity of superoxide dismutase(SOD)increased slightly under 30 ℃ and decreased under 38 ℃. The variational extent of SOD and CAT had positive correlation with the heat tolerance and the increase extent of H2O2 and MDA had the negative correlation under high temperature. The increase extent of Pro had negative correlation with the heat tolerance under 30 ℃ and positive correlation under 38 ℃. There was weak correlation between the increase extent of APX and the heat tolerance. The comprehensive evaluation of subordinate function and cluster analysis indicate that the heat tolerance sequence of five Rhododendron species was R.mucronatum＞R.molle and R.moulmainense＞R.chihsinianum＞R.rubiginosum，in accordance with their behavior in the field.

Effect of two light irradiances on physiological response of Paulownia fortunei seedling roots to Pb stress

2015/12/16 11:38:50

The physiological responses of P.fortunei seedling roots to Pb stress under natural light and 50% natural light treatments were investigated. Results showed that production rate of superoxide anion，hydrogen peroxide and MDA contents consistently increased with the increase of added Pb. Compared to natural light treatment，their increasing trends were less sharper under 50% natural light treatment. Under natural light treatment，the activities of SOD，CAT and APX increased firstly and then decreased with the increase of added Pb，while the activities of POD consistently increased. At 50% natural light treatment，all the antioxidant enzyme activities consistently increased with the increase of added Pb. The activities of SOD and POD under 50% natural light treatment were higher than those under natural light treatment. Results suggested that the decrease of light irradiance relieved the oxidative damage of Pb stress to the roots of P.fortunei.

SPDinduced salt resistance and mechanism in Chinese cabbage

XU FenFen — 2015/12/16 11:38:50

We studied the effects of exogenous SPD of different concentration 0，50，100，150，200 mg/L on the physiological parameters in Chinese cabbage by the method of rhizosphere and foliar spraying. The results were as follows:SPD of 100-150 mg/L significantly improved the plant height，plant dry weight，plant fresh weight and plant water content，reduced the malondialdehyde(MDA)content. enhanced superoxide dismutase(SOD)，peroxidase(POD)enzyme activity of leaf，significantly increased the chlorophyll content and the net photosynthetic rate of Chinese cabbage under the salt stress.

Effects of nitrogen fixing bacteria on photosynthesis and chlorophyll fluorescence in sugarcane at elongating stage

2015/12/16 11:38:50

Use of biological nitrogen fixing organisms is an effective approach to reduce the high cost of nitrogen fertilizer in sugarcane production system. The present experiment was conducted to investigate the effects of using nitrogen fixing bacteria(strain A01) on photosynthetic characteristics of different sugarcane genotypes during elongating stage. Six sugarcane varieties，viz.，GT24，GT9，YC84/153，GT18，CP65/357 and F172 were grown under nitrogenfree conditions in greenhouse. The endophytic nitrogen fixing bacteria was inoculated to sugarcane plants during seedling stage. And the chlorophyll content，photosynthetic rate(Pn)，intercellular CO2 concentration(Ci)，stomatal conductance(Gs)，transpiration rate(Tr)，maximum quantum yield of PSII(Fv/Fm)and actual photochemical efficiency of PSII(φPSIIR) were measured in +1 leaves of sugarcane during elongating stage. The results showed that the nitrogen fixing bacteria increased the chlorophyll content，Pn，Gs and Tr in the leaves of all the six sugarcane varieties. The chlorophyll fluorescence parameters such as Fv/Fm ratio was regulated in certain degree under dark adaptation，but φPSIIR was significantly enhanced in plants inoculated with the bacteria. The results suggested high utilization efficiency of nitrogen fixed by the endophytic bacterium.

Dynamic changes of stomatal density and stomatal index during the leaf development in Catharanthus roseus

YANG Ying， WANG YongFei， MA SanMei* — 2015/12/16 11:38:50

The present paper focused on dynamic changes of stomatal density and stomatal index during the developmental stage of Catharanthus roseus leaves. It was found that at all developmental stages，stomata mainly distributed along veins on adaxial surface，while uniformly distributed on abaxial surface of the whole blade. The ten pairs of leaves numbered from bottom to top in one shoot were divided into ten developmental stages which were numbered from top to bottom，e.g.leaves in the 10th nod were in stage Ⅰ，the rest might be deduced by analogy. Due to cell division and cell enlargement during the first three stages，the leaves of C.roseus grew rapidly. In stage Ⅲ，the leaf length could reach (4.5±0.4) cm. Cell division and cell enlargement basically stopped at stage Ⅸ. Stomatal density and stomatal index kept stable from stage Ⅸ to stage Ⅷ. In the last two stagesstage Ⅸ andⅩ，stomatal density and stomatal index increased abruptly.

Adventitious buds formation from immature or near mature embryo cultures of Ginkgo biloba

CHEN Ying， XU CaiPing， SHENG LiLi, LI ShuXian， CAO FuLiang* — 2015/12/16 11:38:50

The formation of adventitious buds of Ginkgo were studied by using near mature and immature embryos cultures. The results were as followed:(1)Ginkgo embryo had characteristic of physiological afterripening. The rate of small cotyledon embryos(1.0-3.0 mm) was 26.1%，the rate of large cotyledon embryos(3.0-5.0 mm)was 422% and the total rate of the seed with embryo was 71.1% in October； (2)The cotyledons from large cotyledon embryos became swollen and few callus was produced after cultured on different induction mediums，but the small cotyledon embryos of all could induced callus and the green spots were initiated on the surface of callus after subculuring two times； (3)The adventitious buds formed from cotyledon of large cotyledon embryos after subcultured on modified MS medium with 0.2 mg•L1 IAA and 0.75 mg•L1 KT. The highest emergence of adventitious buds was 20.0%； (4)The highest frequency of embryo bud germination of near mature embryos (42.5%)and the highest number of bundle buds per explant(3.0)were obtained on modified MS medium with 0.01 mg•L1 NAA and 1.0 mg•L1 6BA.

Influencing factors of efficient and rapid propagation in vitro of Bromeliads

2015/12/16 11:38:50

This paper，every stage was studied by using lateral buds as explants. The results showed that the variety had remarkable influence on the contamination rate and the browning rate of explants，and the induction rate and the proliferation multiple of tufty buds. The variety and concentration of exogenous hormone were the major influencing factors in every stage such as the bud induction, proliferation and the root induction. In the bud induction stage，it got the best result of induction differentiation，which occured after 48 days with 40% induction rate and average six buds per explant when they were cultured on 1/2MS basal media supplemented with NAA 0.2 mg•L1 and 6BA 2 mg•L1. In the bud proliferation stage，when the concentration of 6BA solely supplemented in the media between 0-4 mg•L1 was higher，the proliferation multiple was higher. When the concentration of NAA solely supplemented in the media was 0.2 mg•L1，the proliferation multiple was the highest. It got better proliferation on the media supplemented with both 6BA and NAA than that on the media supplemented with 6BA or NAA solely. On MS media supplemented with 3 mg•L1 6BA and 0.5 mg•L1NAA，the proliferation multiple of G.dissitisflora and G.‘Claret’both got to the top，which was 5.24，3.84 respectively. On any the same media，the proliferation multiple of G.dissitisflora was remarkable higher than that of G.‘Claret’. It was better to root when the plantlets grew on 1/2MS media containing 0.5 mg•L1 NAA and 0.5 mg•L1 IBA，and the rooting rate could reach 91.03%，and the average number of the roots was 5.3 piece per plant.

Effects of aluminum stress on growth characteristics and biomass allocation of Plantago virginica

CHEN Xiang1， LU YaoDong1*， HUANG Wei2， HU XueHua2 — 2015/12/16 11:38:35

The objective of the present investigation is to observe the effects of different Al3+ concentration (100，500，800，2 000 mg/L) along with different duration times (10，20，30 d) on growth characteristics and biomass allocation of the invasive plant P.virginica. The results showed that 100 mg/L of Al3+ had little effect on the growth of P.virginica. Under 2 000 mg/L of Al3+，both leaf areas and leaf numbers of P.virginica decreased. Spike number of P.virginica was promoted under 2 000 mg/L Al3+ stress for a short treatment time，but its growth velocity was inhibited over a long period 2 000 mg/L. With aggravating Al3+ stress，a trend was found that its biomass allocation prefered aboveground to underground. The proportion of its spikes increased rapidly to a peak and then declined rapidly over the subsequent days under high concentration of Al3+，revealing an adaptive strategy for its reproducing during unfavorable circumstances.

Absorption of exogenous iodine in rhizosphere and its effects on physiological parameters of cherry tomato plants

YU WenJin， YAO Yan， WEI HuiMing， LONG MingHua， TANG XiaoFu — 2015/12/16 11:38:35

Cherry tomatoes(Lycopersicon esculentum var.cerasiforme)were cultured in deep flow hydroponic solution supplemented with graded levels(1.0-6.0 mg•L1)of iodide(I). The results showed that the iodine content in roots，leaves and fruits increased with increasing of I concentration. After I treatment，initially，the iodine content in roots increased rapidly for the first week；a decrease in the second week followed；and afterwards，a tendency of gradual increase was observed，with more significant changes at higher concentrations of I(3.0-6.0 mg•L1) as compared to lower ones(1.0-2.0 mg•L1). During the first three weeks after the I treatment，the iodine content in leaves increased significantly with the passage of time；after the third week，the increase tended to slow down for the plants treated with high concentrations of I(4.0-6.0 mg•L1)，while no significant changes were found at medium or low ones(1.0-3.0 mg•L1). For plants with I treatment of 1.0 mg•L1，the iodine content in roots was higher than that in leaves；for I treatment of 2.0-6.0 mg•L1，the iodine content in roots was higher than that in leaves at the early growth stage，but lower than leaves at the later growth stage，which suggested that iodine did not transfer to leaves until it had accumulated in the roots to a certain amount. Making monofactor and twofactor regression analyses respectively of the correlation between the iodine content in ripen fruits and that in the leaves and the roots after I treatments of 1-6 weeks，we found that it was the first week after I treatment that the iodine content in fruits was affected most，and the iodine content in ripen fruits could be predicted with the iodine content in leaves and roots using the dualistic linear regression equation. The I concentration of 2.0-6.0 mg•L1 significantly affected the deoxidize amount of triphenyl tetrazolium chloride(TTC) in roots，and the influence mainly appeared within the first three weeks after adding I to the solutions. The low concentrations of I(1.0-2.0 mg•L1) did not affect chlorophyll content significantly，but the medium or high concentration of I(3.0-6.0 mg•L1) significantly reduced it. Two weeks after treatment with I concentration of 2.0-6.0 mg•L1，the photosynthetic rates(Pn)，stomatal conductance(Gs)，intercellular CO2 concentration(Ci)，transpiration rates(Tr)significantly reduced，while the stomatal limitation values(Ls) increased significantly.

Responses of Mikania micrantha to parasitization of Cuscuta campestris in total soluble protein content and activities of antioxidant enzymes

2015/12/16 11:38:35

To develop efficient biocontrol techniques for the invasive weed Mikania micrantha，the use of the obligate parasitic plant，Cuscuta campestris Yuncker as a biological control was investigated. In this experiment，whether the impacts of the parasite on host soluble protein content and activities of some antioxidant enzymes were affected by the density of the parasite was tested. The responses of M.micrantha to parasitic densities of 0，1，2，4 and 8 individual seedlings of C.campestris per host plant were examined.On the 30th day of parasitization，infection with more than 1 C.campestris seedling significantly lowered soluble protein content. Compared with control，activities of superoxide dismutase (SOD) and peroxidase (POD) significantly increased at parasitic density of 1 parasite per host plant；but as parasitic density increased，both SOD and POD activities decreased.Moreover，SOD activity was not changed but POD activity was significantly decreased at parasitic density of 4 parasites per host，and both SOD and POD activities were significantly decreased at parasitic density of 8 parasites per host，compared with control. The infected plants had significantly lower catalase (CAT) activity but higher SOD/CAT，SOD/POD and SOD/ (CAT+POD) ratios than the control at all parasitic densities. The results indicated that the effects of C.campestris infection on M.micrantha were density dependent，which provided a basis for refining this strategy for biological control of M.micrantha. The optimal costeffective number of parasites to control M.micrantha was 4 per host plant in the field.

Studies on isozymes during somatic embryogenesis of Tapiscia sinensis

ZHANG Bo， JING DanLong， LI XiaoLing，WANG YuBing, ZHANG DeChun* — 2015/12/16 11:38:35

Polyacrylamide gel electrophoresis was used as a tool to analyze isozyme patterns such as peroxidas〖JP3〗e(POD)，esterase(EST)，amylase(AMY)and superoxide dismutase(SOD)in somatic embryogenesis of Tapiscia sinensis. The results showed that a higher expression of EST，POD，AMY and SOD isoenzymes indicated that there was a higher physiological metabolic rate in the stage of globular embryos，and SOD became more which might be associated with the processes of somatic cells from the differentiation into embryonic regeneration. POD continued to change and strengthen in the process of somatic embryogenesis；At the same time，the expression of AMY also was strengthened. The activity variations of POD，EST，AMY and SOD were closely related to the induction of embryogenic callus and the development of somatic embryos in somatic embryogenesis of T.sinensis. These〖JP2〗 enzymes could be used as the signs during the development of somatic embryos，and would be as a reference label of the specific developmental stages.

Isolation and identification of pathogens that caused Curcuma alismatifolia anthracnose

2015/12/16 11:38:35

The pathogens that caused Curcuma alismatifolia anthracnose were isolated using usual tissue isolation. Strains were cultivated，purified，reinoculated in the field and laboratory. The strain that caused the anthracnose was reisolated. Morphological and molecular biological techniques were used to identify the strain. Results showed that 6 strains were isolated from infected leaves of C.alismatifolia，but only Cum3 caused the anthracnose. The incidences of Cum3 disease reinoculated in the field and laboratory were 96.7% and 100% respectively. Cum3 is a kind of fungi belonging to the genus Fusarium，and its colonial morphology is a circular form，with white mycelium，diaphragms and branches. There are three kinds of conidia:microconidia，macroconidia and chlamydospores. At last，the sequence of internal transcribed spacer (ITS) region of Cum3 was analyzed，the length was 528bp and the sequence was compared with other species in the Genbank. The symptom in the field indicates that the pathogen of Curcuma alismatifolia anthracnose is Fusarium proliferatum.

Study on the development correlation between male and female gametes and the induction of 2n gamete of Rongan kumquat （F.crassifolia）

2015/12/16 11:38:35

With Rongan kumquat(F.crassifolia)as the experiment material，the process of meiosis and the development of microsporocyte，female gametophyte and the natural incidence of 2n pollen were observed and buds were induced mutation with colchicine of different concentrations and times. The main results showed that:(1)The proportion of 2n pollen of Kumquat was 0.31% under natural state. (2)When the longitudinal diameter of the bud was 0228-0270 cm，the microsporocyte was premeiosis Ⅰ；When the longitudinal diameter of the bud was 0274-0318 cm，the megaspore mother cell was in the early stage of meiosis，and the microsporocyte was in the stage from meiotic metaphase to tetrad，male and female gametes did not develop synchronously. (3)The induction rate of 2n female gametes was 9.09% when treated with colchicine of 0.25% for 2 days. (4)0.67% of 2n pollen was observed when treated with colchicine of 0.30% 4 days after treated. It was found that 2n pollen germinated 3 hours later than n pollen and 2n pollen tube growth rate was also lower.

Study on embryo rescue of interspecific hybrids between Hedychium

XIONG YouHua1，2， ZHUANG XueYing2， LIU Nian1* — 2015/12/16 11:38:35

The object of this study was to improve the germination capacity of embryoes of Hedychium hybrids through embryo rescue in vitro. Hybrid ovules of H.coronarium×H.gardnerianum were used to investigate the effect of different growth stage of embryo，different culture medium and lowtemperature treatment on the germination capacity of embryoes. The results indicated that MS medium supplemented with 0.1 mg/L BA and 0.1 mg/L NAA showed suitable for embryo rescue in vitro culture. It was showed that the optimum germination capacity of embryoes reached 40.6%，which was obtained by taking 60 day old immature embryo as explant. Lowtemperature treatment of fruits for 3-6 d，had a positive effect on improvement of the germination capacity of embryoes.

Studies on anther callus induction of rice by orthogonal test

WANG Hui， XIAO XiaoJun， QI ZeMin*， HUANG ZuoXi， WANG Fang — 2015/12/16 11:38:35

Using the anther of Milyang 46 and the F1 hybrids of Pei’ai 64/8006，the correlation of pollen development with the distance of the pulvinus was studied. The results showed that most of the microspores were in the late uninucleate stage when the distance of the pulvinus was 5-7 cm for Milyang 46；while it was 7-9 cm for the F1 hybrids of Pei’ai 64/8006. Besides，the effects on anther callus induction rate of the types of medium，the ration of plant hormones，the concentration of the sucrose and 2，4D were investigated by using orthogonal tests L9(34)，and the best conditions for callus induction of the two accessions were selected:the Milyang 46 was SK3+4 mg•L1 2，4D+1 mg•L1 NAA+0.5 mg•L1 KT+3%sucrose while the F1 hybrids of Pei’ai 64/8006 was HE5+2 mg•L1 2，4D+1 mg•L1 NAA+0.5 mg•L1 KT +6%sucrose.

Dynamic changes in endogenous hormones in Taxus chinensis var.mairei seed during stratification

2016/1/15 3:04:59

In order to characterize relationship between endogenous phytohormones and dormancy of Taxus chinensis var.mairei,changes in contents of four kinds of endogenous phytohormones,i.e.,abscisic acid(ABA),gibberellin(GA3),indole acetic acid(IAA)and zeatin riboside(ZR)in spermoderm and endosperm of Taxus mairei were analysed by enzyme-linked immunosorbent assays(ELISA). The results showed that ABA contents were gradually decreased in endosperm during stratification,while GA contents were increased and IAA and ZR contents firstly increased and then decreased. Calculated ratio data of GA/ABA,IAA/ABA and ZR/ABA exhibited gradually increasing trends within stratification time and dormancy was relieved accordingly with them. In spermoderm,endogenous ABA,GA,IAA and ZR contents were gradually decreased through stratification. The change in GA/ABA ratio was greater than the other two ones. So it was suggested that the tradeoff between ABA and GA contents could control the dormancy process in T.chinensis var.mairei seed.

Toxic of chromium(Cr⁶⁺)on the callus of Alternanthera philoxeroides

2016/1/15 3:04:59

This study focused on changes of several representative physiological characteristics and ultrastructure in the callus of Alternanthera philoxeroides under different concentrations of Cr⁶⁺ stress(0,0.1,0.2,0.5,1.5 mmol·L^-1). The results showed that with the increasing of Cr⁶⁺,(1)total chlorophyll(Chl),chlorophyll a/b(Chl a/b)and total soluble protein contents showed the same trend,which increased firstly and then decreased.(2)Superoxide anion(O^{^</sup>₂)and hydrogen peroxide(H₂O₂)content irritably increased compared to the control group,then decreased,however,it's always higher than that of control.(3)The soluble sugar content enhanced with the increasing of Cr⁶⁺ concentration.(4)Activities of supeoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)and ASA and GSH contents all increased first and then decreased.(5)Transmission electron microscope observation indicated that Cr⁶⁺ also imposed injury action on ultrastructure of callus cells:thylakoid twisted,chloroplast envelope broke and large starch grain formed,mitochondria disappeared and formed vacuolization,nucleolus lost and chromatin condensed in nucleus,and endoplasmic reticulum vanished by means of vesiculation. The conclusion could be reached that toxic effects of Cr⁶⁺ on callus resulted in disorder of physiological and biochemical metabolism and destruction of ultrastructure of callus cells.}

Cloning and sequence analysis of chalcone synthase gene Chs from Fagopyrum esculentum

2016/1/15 3:04:59

The cDNA ORF sequence of chalcone synthase gene(FeChs)was amplified by RT-PCR from Fagopyrum esculentum. Its accession number is GU172166.1 on GenBank. The anlyasis indicated the cDNA sequence was 1 179 bp,encoding 392 amino acids and had a homlogy 78%-92% with other plants'CHS genes. The deduced amino acids sequence of FeChs contained the highly conserved catalytic active sites,and the typical Tag sequence of CHS with GFGPG.

Construction of SSH-cDNA library from different developmental stages of Siraitia grosvenorii

2016/1/15 3:04:59

In order to reveal the differentially-expressed genes involved in triterpenoid saponin biosynthesis of Siraitia grosvenorii,50 DAF(days after flowering)and 70 DAF fruits from different developmental stages were employed to construct subtractive cDNA library by suppression subtractive hybridization(SSH)in this study. Total 641 positive clones were selected randomly and sequenced from the forward-subtracted cDNA library(70 DAF as the tester and 50 DAF as the driver),and 622 high quality sequences were obtained. The rate of the recombination fraction was above 96%. The distribution of inserted fragments ranged from 101 bp and 934 bp,and the average fragment size was 500 bp. Followed by BLASTN and BLASTX for sequences dates,201 ESTs showed no significant matches to any other sequences in NCBI nucleotide sequence database,they were probably novel genes. The other 421 ESTs carried with remarkable identity to proteins with known function in the database,which fell into several functional categories including energy and secondary metabolism,transcription factors,ripening,senescence and pathogen-resistance. Further results indicated that the cDNA library quality were conformed to SSH library standard and could be used in further studies,also would provide reference for studying the differentially expressed genes,exploring molecular mechanism of triterpenoid saponin biosynthesis,and increasing productivity and quality of mogrosides in S.grosvenorii.

Effects of arbuscular mycorrhizal fungi on the rooting, growth and enzymatic activity relating to disease resistance of beach plum(Prunus maritima)cuttings

2016/1/15 3:04:59

The effect of inoculation with Glomus mosseae,G.diaphanum or G.etunicatum on the rooting,growth and enzymatic activity relating to disease resistance of beach plum cuttings were studied. The results showed that the largest percentage rooting(47.6%),the maximum number of lateral fine roots(20.4),the largest dry weights(DWs)of roots(0.26 g),the largest DWs of shoots(3.55 g),greatest heights of cuttings(51.3 cm),and the largest total leaf areas of cuttings(1 012.9 cm²)were observed following inoculation with G.mosseae. The greatest root lengths were observed alter G.etunicatum inoculation(19.3 cm). Except for the numbers of primary root generated,G.mosseae and G.etunicatum inoculations were significantly better than the controls in all the parameters studied. As for the numbers of primary roots,inoculation with AM fungi gave values close to the control. Greater uptake of macronutrients(P,K,Mg,and Ca)and micronutrients(Mn,Cu,Zn and B)uptake was observed in beach plum for all three Glomus species 80 d after inoculation. Among the three Glomus species,G.mosseae was most effective,G.etunicatum next and G.diaphanum gave macronutrient uptake values close to controls. Compared with controls,the activity of POD,polyphenolase and PAL were increased after inoculation with all three Glomus species. Among the three Glomus species,G.mosseae was most effective,G.etunicatum next and G.diaphanum was close to controls. This study showed the beneficial effects of inoculation with each of three Glomus fungi,especially G.mosseae and G.etunicatum,on inducing the rooting,growth and enzymatic activity relating to disease resistance of beach plum cuttings.

Effects of high temperature on key enzymes activity of nitrogen metabolism in flag leaf and protein content in grain after flowering of early-indica rice

2016/1/15 3:04:59

Estigate the effects of high temperature on key enzymes(GS and GOGAT)activity of nitrogen metabolism in flag leaf and protein content after flowering in two different early indica rice,high temperature(31.5 ℃)and optimal temperature(23.5 ℃)are treated to two different type early indica rice(Xiangzaoxian 24 and Zhuliangyou 611). The results showed that the GS and GOGAT activity were highest on 10 d under high temperature treatment and were slowly decreased under optimal temperature treatment. Activity of two kinds of key enzyme in Xiangzaoxian 24 were both higher than that in Zhuliangyou 611. Compared to optimal temperature treatment in Xiangzaoxian 24 under high temperature,the protein content was 4.75 mg higher on 22 d and 1.88mg higher for Zhuliangyou 611. These results suggest that high temperature can help to improve two early indica rice(especial the Xiangzaoxian 24)the GS and GOGAT activity in flag leaf and grain protein content.

Studies on callus induction and gametophyte regeneration of Bartramia pomiformis

2016/1/15 3:04:59

This research was conducted to select a suitable culture system for the callus induction and gametophyte regeneration of Bartramia pomiformis. And axenic mosses were inoculated on MS and Knop solid media with different hormone combilations for callus and buds induction and differentiation. The result showed that MS medium containing 0.5 mg/L BA,0.1 mg/L 2,4-D was optimal for callus induction,with the induction rate of 33.3%. Optimal medium for inducing gametophyte is MS+1.0 mg/L BA,and the achieved multiplication coefficient was 10.0. And Knop with no hormone was as well as MS in generation of gametophyte. This investigation could provide some important references for callus induction and systematics of B.pomiformis.

Perspectives and research advances on the Thermogenesis

2016/1/15 3:04:59

Compared with the international research,the floral thermogenesis study is still at its initial stage of development. Few studies in this field have been reported in China. Based on overseas researches in this field and unpublished data from our research group,the concept of floral thermogenesis is summarized and put forward for the first time in domestic research field. Floral thermogenesis could be defined as:the flowers in some plant taxa produce amount of heat actively,and maintain remarkably constant temperatures in face of fluctuating ambient temperature. It is an active process of heat production and favorable for plant reproductive development. In this paper,existing researches in this field are summarized and the latest advances from phenomenon observation to the internal control mechanism are reviewed.

Progress on nitrogen uptake and transport in plant

2016/1/15 3:04:59

Nitrogen is a major nutrient element which plays a key role in plant growth and development. Nitrogen is available to plants both in the forms of nitrate and ammonium. A different molecular mechanism of nitrogen in their uptakes and transports is formed in plant evolution. In this paper,the physiological characteristics,molecular mechanism and relevant genes of nitrogen uptake and transport in plant are summarized,which may provide theoretical principles for the genes related to the nitrogen uptake and transport in rice.

Effects of media temperature on rooting and leaf photosynthesis of Bougainvillea glabra cuttings

XIE Zhi-Nan, ZHONG Zan-Hua, LAI Rui-Yun, ZHANG Xue-Qin, SU Ming-Hua — 2016/1/15 3:04:42

The effects of media temperature(18 ℃,20 ℃,25 ℃,30 ℃ and 33 ℃)on rooting and leaf photosynthesis of Bougainvillea glabra cuttings were measured under the condition of intelligent monitor and control. The results showed that with the increasing of media temperature,an increasing of the net photosynthesis rate,transpiration rate,stomatal conductance and intercellular CO₂ concentration in B.glabra cuttings leaves,while the stomatal limitation decreased. The net photosynthesis rate was lower and there was no obvious differences of treatment 18 ℃ and 20 ℃. The differences of net photosynthesis rate among treatment 25 ℃,30 ℃ and 33 ℃ were obvious. The rooting number per cutting was less,and the root length was shorter,while the survival rate of the cutting was lower when the media temperature was lower than 20 ℃. When the media temperature was higher than 30 ℃,the fallen number and the basal stems rotting rate of cutting increased,while the survival rate decreased. In summary,25-30 ℃ is the optimal media temperature for rooting of Bougainvillea glabra cuttings.

Effects of aluminum stress on growth and characters of photosynthesis and transpiration in polygonaceae plants

LIU Qiang, YIN Li, LONG Wan-Wan, XIAO Yi-An — 2016/1/15 3:04:42

Three polygonaceae plants(Polygonum lapathifolium,P.perfoliatum and P.hydropiper)were used to study the effects of five aluminum(Al)concentrations(0,25,50,100,400 μmol·L^-1)on leaf photosynthesis,transpiration and chlorophyll fluorescence characteristics with the solution culture method. The results showed that the high Al treatment(400 μmol·L^-1)significantly decreased the growth of aboveground and root system,the leaf chlorophyll content,Chla/Chlb,net photosynthetic rate(Pn),water use efficiency(WUE),quantum yield of electron transport at PSII(φPSII)and coefficient of photochemical quenching(qP)of three polygonaceae plants. In the low and medium concentrations of Al treatments(25～100 μmol·L^-1),the biomass was significantly promoted in P.lapathifolium,significantly reduced in P.perfoliatum,promoted firstly and then reduced again in P.hydropiper compared to the control. Meanwhile,the chlorophyll content,Chla/Chlb,Pn,transpiration rate(Tr),intercellular CO₂ concentration(Ci),maximal photochemical efficiency(Fv/Fm)and qP were not significantly changed in P.lapathifolium and P.hydropiper,but WUE,φPSII and coefficient of photochemical non-quenching(NPQ)were significantly decreased in P.hydropiper only. However,in P.perfoliatum,leaf photosynthesis,transpiration and chlorophyll fluorescence characteristics other than Ci,Fv/Fm,were decreased remarkably. These results indicated that P.lapathifolium could enhance the tolerance to Al stress under low and medium Al treatments by maintaining steady chlorophyll content,Chla/Chlb,WUE,Pn,photochemical reaction efficiency at PSII and increasing non-radiative dissipation of absorbed excitation energy through NPQ.

Esterase isozyme on sprouting seeds of genus Fagopyrum

2016/1/15 3:04:42

The esterase isozyme of thirty-three collections of cultivated and wild buckwheat belonging to eight species of genus Fagopyrum including seven species in the big-achene group and one species in the small-achene group were studied by means of polyacrylamide gel eletrophoresis(PAGE). The results showed that there were twenty-two different bands of esterase isozyme,with a range from four bands to eight bands among different buckwheat species. F.esculentum and F.tataricum had eight bands and seven bands,respectively. The variations of esterase isozyme zymograph were much great among different buckwheat species but less in the same species. The results of the zymograph analysis and the clustering showed that there were great differences of zymographs between the big-achene group and the small-achene group and that F.megaspartanium and F.pilus were close in clustering tree to F.esculentum and F.tataricum,respectively,supporting Chen's hypothesis that F.megaspartanium and F.pilus might be the ancestor species of F.esculentum and F.tataricum,respectively.

Genetic diversity in kenaf(Hibiscus cannabinus) UG93 cytoplasmic male sterile lines,maintainer lines and restorer lines revealed by ISSR and cpSSR markers

2016/1/15 3:04:42

8 ISSR primers and 7 chloroplast SSR(cpSSR)primer pairs were employed to study genetic diversity among 9 pairs of kenaf UG93 cytoplasmic male sterile/maintainer lines and 5 restorer lines. Nucleus genome genetic similarity coefficients(SC)based on ISSR ranged from 0.333 to 1.000,with the average of 0.583,0.689 and 0.812 among maintainer lines,maintainer and restorer lines,and restorer lines respectively. Chloroplast genome SC based on cpSSR ranged from 0 to 0.286. The 23 tested lines were clustered into 4 haplotypes,with all 9 sterile lines were classified into a single type. 5 of 9 couple of sterile/maintainer lines showed some degree diversity which indicated that replacement of nucleus genome between sterile line and maintainer line was not complete. The results will help generating new kenaf sterile resource and excellent cross combinations.

Comparative analysis of three different methods of total DNA extraction used in Taxus

2016/1/15 3:04:43

Species of Taxus in China are endangered and were listed as the national first-class protected wild plant. In present study,three different methods of total DNA extraction,i.e.SDS,CTAB and DNA extraction Kit(DNeasy Plant Mini Kit),were used to isolate genomic DNA from dried leaf materials of Taxus. The total genomic DNA yielded by the three methods were quantified and analyzed by spectrophotometer,agarose gel electrophoresis and PCR reaction respectively. The results of the three methods of DNA extraction were compared and analyzed,which indicated that the CTAB method could yielded relatively pure and high amount total DNA and was highly suited for use directly in downstream applications. The method of DNA extraction using Kit(DNeasy Plant Mini Kit)also could yield high quality total DNA with the highest purity of the DNA comparing to other methods used in this study. However,the relatively lower DNA yield and higher costs of this method might be limited to its wide use. Whereas the SDS method could not yield high quantity of total DNA in most sampled accessions in the present study,which indicated low purity of the yielded DNA of SDS method could not be used in downstream application directly. We also discussed the factors influencing on the total genomic DNA yield and purity,which could help to improve and select the suitable protocol of DNA isolation of plants.

Transferring disease resistance gene NPR1 into Siraitia grosvenorii

2016/1/15 3:04:43

Using cotyledon of Siraitia grosvenorii as explants,an efficient and reliable transformation system was established. Several factors affecting genetic transformation of Agrobacterium-mediated S.grosvenorii were studied. The results showed that 5 days aged cotyledon using for infection explants,pre-culture 1 d,bacterial infection time 20 min,co-culture 4 d with 22 ℃,AS concentrate 100 μmol/L,10 mg/L of hygromycin was suitable screening concentration for the differentiation of adventitious bud and root could get the better transformation efficiency under 20 mg/L of hygromycin. The transformants were confirmed by PCR and Southern blotting analysis with the transformation rate 1.5%.

Cytological observation of embryo sac and embryonic development in the new mandarin cultivar ‘Wuzibayueju'(Citrus reticulata Blanco)

2016/1/15 3:04:43

Enzymolysis oscillation squash method and routine paraffin section were used to study embryo sac fertility of ‘Wuzibayueju' and the embryonic development of three pollinated combinations(‘Wuzibayueju'×‘Wuzibayueju',‘Wuzibayueju'×‘Ponkan',‘Wuzibayueju'×‘Bayueju'). The results were as follows:(1)The embryo sac fertility of ‘Wuzibayueju' was normal. Mature embryo sac had one egg,two synergids,three antipodal cells and a big central cell containing two polar nuclei.(2)The embryonic development of self-pollinated ‘Wuzibayueju' and cross-pollinated ‘Wuzibayueju'×‘Ponkan',‘Wuzibayueju'×‘Bayueju'were normal. Two weeks after self-pollination,the zygotic embryo became globular-shaped and a few heart-shaped. Three weeks later,heart-shaped embryos and torpedo-shaped embryos were observed. Four weeks later,embryo reached at the torpedo stage. Five weeks later,cotyledonary embryos were observed. And seven weeks later,cotyledonary embryos were developing into seeds while funiculi were still existent. Those results indicated that embryo sac fertility of ‘Wuzibayueju' was normal and there was no embryo abortion during embryonic development. Therefore,the embryo abortion was not the seedless reason of the new cultivar ‘Wuzibayueju'.

Tissue culture of Impatiens blepharosepala and ultrastructural observation of callus

2016/1/15 3:04:43

Leaves,stem and root of Impatiens blepharosepala seedling were cultured in vitro,and two types of callus having different differentiation were observed by electron microscope. The results showed that the induction rate was highest on MS+NAA 0.5 mg/L+6-BA 0.5 mg/L,the induction rate of leaves was 100% and the rate of stems was 67%,but the roots can't be induced. Callus of leaves can be induced buds on MS+NAA 0.5 mg/L+6-BA 2.0 mg/L after 40 d,but there was no organ generated from stems. Cells of callus having differentiation arrayed irregularly and there was obviously similar tissue differentiation by electron microscope.

Comparison of the physiological characteristics of paper mulberry(Broussonetia papyrifera) and mulberry(Morus alba)under simulated drought stress

2016/1/15 3:04:18

Under the treatment of different concentrations of polyethylene glycol(PEG 6000),the carbonic anhydrase activity,photosynthetic responses to light and CO₂,and chlorophyll fluorescence characteristics in leaves of two woody plant species,paper mulberry(Broussonetia papyrifera)and mulberry(Morus alba),were determined. The drought resistance of the two species plants was compared. The results indicated that the activity of carbonic anhydrase in paper mulberry did not vary significantly. There are significant differences of the activity of carbonic anhydrase in mulberry among treatments with different concentration of PEG 6000. PEG 6000 had little influence on net photosynthetic rate of paper mulberry,but caused significant inhibition on mulberry. No significant variation of maximum quantum yield of photosystem II(Fv/Fm)and actual photosynthetic efficiency of PS II(ΦPSⅡ)of paper mulberry among the treatments of PEG was observed,but those of mulberry were significantly inhibited by PEG. Fv/Fm and ΦPSⅡ of paper mulberry was also higher than those of mulberry. The photochemical quenching coefficient(qP)and electron transport rate(ETR)of paper mulberry was significantly higher than those of mulberry. The stable carbonic anhydrase and the strong photosynthetic capacity is an important mechanism for paper mulberry to adapt to karst-arid environment.

Effects of iron stress on growth, physiological character and iron distribution of the three citrus rootstocks

2016/1/15 3:04:18

The three citrus rootstock seedlings such as trifoliate orange,sour orange and red tangerine were taken as experimental material,the effects of iron stress on the growth,physiological character and iron distribution of the three rootstocks were investigated with hydroponics. The results showed that the growth index and chlorophyll concentrations in leaves of the three rootstocks were significantly lower in the iron deficiency(0 μmol·L^-1)than in the treatments of low iron(5 μmol·L^-1)and optimal iron(50 μmol·L^-1),the activities of POD,CAT in leaves and roots decreased and the proline content in roots increased significantly under the iron deficiency treatment,and the activities of SOD in leaves and roots of the three rootstocks changed irregularly. Under the treatments of iron deficiency and low iron,iron contents in roots showed no difference among the three rootstocks,but were significantly lower than those of optimal iron treatment,respectively. The leaf/root and the stem/root of Fe translocation coefficient in the three rootstocks decreased with increasing supply of Fe,whereas no significant difference was found in the leaf/stem of Fe translocation coefficient.

Evaluation on soil pollution in Pingle Manganese Mine of Guangxi using Vicia faba root-tip cells micronucleus testing

2016/1/15 3:04:18

In order to study the pollution status of soil in Pingle Manganese Mine wasteland of Guangxi,four sampling plots respectively in peanut field,persimmon field,orange field and peach field were selected and investigated,the PI and MCN‰ of root-tip cells under soil extracts from all plots were determined. By using micronucleus test technique of Vicia faba root-tip cells,this paper evaluated the pollution status of soil in Pingle Manganese Mine wasteland of Guangxi,and analyzed the reasons that might cause soil pollution. The results showed that the MCN‰ of root-tip cells of V.faba treated with the soils by means of maceration was obviously higher than that of the cells treated with the negative control group(P<0.05); orange field was under the most serious pollution,the PI had reached up to 2.93,while peanut field got the least,the PI was 1.97. In conclusion,these indicated that the soil in this wasteland was more or less polluted,suggesting that it's not suitable for crops at the early stage of farming.

Glutamate oxaloacetate transaminase isozyme in 10 species of genus Epimedium

2016/1/15 3:04:18

The glutamate oxaloacetate transaminase isozyme in 32 young leaf samples of 10 species of Epimedium(including 7 species of the big-flower group and 3 species of the small-flower group)were studied by means of polyacrylamide gel eletrophoresis(PAGE). The results showed that 15 bands of glutamate oxaloacetate transaminase isozyme were discovered in all accessions,likewise,2-15 bands in different species and 2-10 in different plant individuals,showing that zymographs among species were obviously different. Therefore,this could be an auxiliary evidence for the species identification.

Study on the construction of antisense expression vector of Glycine max oleic acid desaturase gene and its transfer to Nicotiana tabacum

2016/1/15 3:04:18

The Glycine max oleic acid desaturase gene fad2-1 was cloned from its genomic DNA by PCR. The amplicon was linked to pMD18-T vector and transformed into E.coli JM109. After sequencing,the correct gene was reversely inserted in pBt expression vector in order to construct plant antisense expression vector,which was transformed into Agrobacterium tumefacien strains LBA4404 by freeze-thawing method. The modified strain LBA4404 was confirmed by double enzyme digestion and PCR detection. The antisense fad2-1 was introduced into tobacco by Agrobacterium tumefaciens-mediated leaf disc transformation,and 75 kanamycin-resistant tobacco plantplets were regenerated. PCR,RT-PCR methods and GC-MS analysis were used to detect the npt-II gene,the transcript of antisense fad2-1 and oleic acid content to get the positive transgenetic plants. The results indicated that the size of the isolated gene was 1196bp,bearing 96.7% identity with the published data in NCBI database. The antisense fad2-1 expression vector was successfully constructed and transformed into Agrobacterium tumefacien strains LBA4404 and tobacco cells,and antisense fad-1 gene was successfully integrated into the genomes of tobacco cells and expressed in transgenic tobacco plantlets.

Effects of He-Ne laser on photosynthesis of wheat seedlings exposed to enhanced UV-B radiation

GAO Li-Mei, LI Yong-Feng, HAN Rong — 2016/1/15 3:04:18

The wheat seedlings(jinmai 8)were exposed to He-Ne laser(5 mW·mm^-2),enhanced UV-B radiation(10.08kJ·m^-2·d^-1)and the combined He-Ne laser irradiation and enhanced UV-B radiation. After being treated for different days(4,5,6,7,8 d)circularly,the changes of the rate of electron transport and relative permeability of chloroplasts membrane,fluorescence emission spectrum were tested with electric conductor and low temperature fluorescence spectrometry. The activities of phosphoenolpyruvate carboxylase(PEPC),carbonic anhydrase(CA),glycolate oxidase(GO),malate dehydrogenase(MDH),chlorophyllase(chlase)and ATPase were measured by photometer. The results showed that the rate of electron transport,the activities of PEPC,CA and ATPase had declined by enhanced UV-B radiation. However,the relative permeability of chloroplasts membrane,the activities of GO,MDH and chlase had increased by enhanced UV-B radiation. Distribution of excitation energy also changed. So the photosynthetic activities of chloroplasts were almost lost. However,the certain dose of He-Ne laser irradiation could partly repaire these damages induced by enhanced UV-B radiation.

Characteristics and dynamic changes of stomata on the hypocotyl of Brassica chinensis var.parachinensis

2016/1/15 3:04:18

The stomata distribution,stomata density,stomata index of Brassica chinensis var.parachinensis on hypocotyl surface were investigated with cultivars“youqing 60 days”and“sijiu-19”by light microscopy. The results showed that ordinary epidermal cells were usually long-flat in shape. Oval-shaped or strip-shaped stomata were observed on hypocotyl epidermis 3 days after seeds germination. The average stomata density on hypocotyl of“youqing 60 days”and“sijiu-19”decreased first and then increased during two years tests. The average stomatal density reached maximum on the 27th day and 21st day respectively. With the growth of seedlings,the average stomatal index on hypocotyl of“youqing 60 days”in the first 24 days increased to a maximum. Comparing with the first 3 days,the difference in stomatal index were significant. The average stomatal index on hypocotyl of “sijiu -19” reached maximum on the first 21 days after germination during the two years experiments,and comparing with that on the 12th day,15th day and 27th day,the difference between them were significant.

Using ¹⁵N isotopic dilution method to quantify the biological nitrogen fixation in sugarcane

2016/1/15 2:42:11

The quantification of biological nitrogen fixation in sugarcane was investigated with ¹⁵N isotope dilution technique in greenhouse bucket culture test,using cassava as reference plant. The experimental results showed that Ndfa accounted for 11.3514%,and the nitrogen fixation rate was 0.9269 g/bucket for whole growth duration of sugarcane. Ndfa and the nitrogen fixation amount in different organs of the plant showed leaf>stalk>root. Ndfa in leaf accounted for 13.2668%,a little higher than that in the whole plant,but the difference was not significant. The percentage of nitrogen from air,fertilizer and soil occupying the total nitrogen of the plant was 11.3514%,7.6857% and 80.9629%,respectively. The nitrogen use efficiency of sugarcane was 58.7583%. It was concluded that all organs of sugarcane could fix nitrogen through biological nitrogen fixation,and leaves could be used to estimate the amount of biological nitrogen fixation in place of whole plant of sugarcane.

Variations of laccase activity and isozymes of Toxico- dendron vernicifluum barkley at tapping season

2016/1/15 2:42:11

It was successful to research on the variations of laccase activity and isozymes of two Toxicodendron vernicifluumsspecies. Barkley varieties which were Gaobachi and Dahongpao in Pingli County of Shaanxi Province were analyzed by spectrophotometry,SDS-PAGE and isoelectrofocusing electrophoresis. The results revealed laccase activity declined from June to July,then rose from June to September and the laccase activity in Gaobachi was higher than Dahongpao at tapping season. Molecular weight was 110 kDa. In T.vernicifluum, there were more than four isozymes which were different between two varieties and it could be a biochemistry index to appraise T.vernicifluum barkley varieties.

Indoor potted plants of Marantaceae and Pteridophy- tes for purification of formaldehyde polluted air

2016/1/15 2:42:11

Ten plants from Marantaceae families and ten plants from Pteridophytes were tested for their abilities of removing formaldehyde(FDH)in the air. Each of the plants was placed for seven days in a 1.0 m×1.0 m×0.8 m glass box filled with FDH with the initial concentration of 15 mg/m³. Some plants such as Neottopteris nidusVolulum,Calathea lubbersiana showed the top resistant ability to FDH; some plants such as Calathea ornata,Calathea setosa,Calathea freddy,and Calathea roseo-picta showed medium resistant ability to FDH; other species such as Neottopteris nidus,Pteris fauriei,Pteris ensiformis cv.Victoriae,Pteris cretica cv.Albolineata,Nephrolepis cordifolia,Cyclosorus parasiticus,Blechnum orientale,Maranta bicolor,and Calathea zebrina showed no resistance to FDH. The absorption of plants for FDH in the glass chamber was found especially apparently different during the first three days. The fastest purification rates of FDH were found in species such as C.zebrina,Microsorum punctatum,and the slowest were found in species such as C.parasiticus,P.ensiformis cv.Victoriae,N.nidus cv.Volulum,and C.setosa. In conclusion,8 species of the potted plants which could be recommendable to be used for FDH purification were N.nidus,Calathea rotundifolia,P.cretica cv.Albolineata,C.ornata,Platycerium bifurcatum,N.nidus cv.Volulum,C.roseo-picta,and C.freddy,because these plants have high absorption ability to FDH and have less damage.

Effects of Cr⁶⁺ stress on 5-MeC content of DNA in wheat seedlings

HEI Shu-Mei — 2016/1/15 2:42:12

The effects of Cr⁶⁺ on the 5-methyl cytosine(5-MeC)contents in leaves and roots DNA in 3-day-old and 10-day-old wheat seedlings with 5-100 mg/L Cr⁶⁺treatment were determined by the reversed-phase high performance liquid chromatography(HPLC). The Hypersil BDS-C18 column(5 μm,150×4.6 mm I.D.)and mobile phase of 5% methanol,-4.75 mmol/L sodium hexanesulfonate, -0.2% triethanolamine-water,pH 5.5 at the flow rate of 0.7 mL/min was used. Detection was performed with UV detector at 273nm. The results showed that the 5-MeC contents in leaves and roots DNA in 3-day-old and 10-day-old wheat seedlings with Cr⁶⁺ concentration increased from 5 to100 mg/L were all higher than that in control,but the 5-MeC contents caused by 100 mg/L Cr⁶⁺was lower than that in control in the roots DNA in 3-day-old seedlings. And the results suggested that 3-day-old seedlings were more sensitive to Cr⁶⁺stress than 10-day-old seedlings, and the roots were more sensitive than the leaves. Therefore,the change of DNA methylation level caused by heavy metal Cr⁶⁺could have an effect on normal growth and development in wheat seedlings.

Germfree germination of seeds and rapid prolife- ration in vitro of Cymbidium floribundum

2016/1/15 2:42:12

Cymbidium floribundum seeds were used to investigate the effects of salt contents,plant growth regulators and light on seed asymbiotic germination. Based on this,the tissue culture and rapid propagation technical system of C.floribundum were established,by investigating the medium formula and culture conditions of proliferation and redifferentiation of protocorm,strong seedlings and rooting of bud. The optimal medium for asymbiotic germination of C.floribundum seeds was 1/6 MS+NAA 0.5 mg·L^-1+6-BA 2.0 mg·L^-1+ meshed potato 50 g·L^-1+AC 1.0 g·L^-1; the light-intensity of culture was 1.25 μmol·m^-2·s^-1; the seed germination rate was 63.6%. The optimal medium for subculture for protocorm multiplication was 1/4 MS+6-BA2.0 mg·L^-1+NAA 0.5 mg·L^-1+AC 1.0 g·L^-1+PE 200 g·L^-1; the proliferation coefficient was 6.5/60 d; the bud redifferentiation rate was 60.2%. The optimal medium for bud redifferentiation was 1/4 MS+6-BA2.0 mg·L^-1+NAA 0.2 mg·L^-1+AC 1.0 g·L^-1+PE 200 g·L^-1; the proliferation coefficient was 4.0/60 d; the bud redifferentiation rate was 85.0%. The optimal medium for strong seedlings and rooting were 1/6 MS+6-BA 3.0 mg·L^-1 +NAA 1.0 mg·L^-1+AC 1.0 g·L^-1+cane sugar 20 g·L^-1+PE 200 g·L^-1 and 1/4 MS+6-BA 2.0 mg·L^-1 +NAA 1.2 mg·L^-1+AC 1.0 g·L^-1+cane sugar 20 g·L^-1+PE 200 g·L^-1,respectively. The rooting percentage was 100%,and transplantsurvival rate reached 90%. This technology could be used to seedling breeding and germ- plasm protection of C.floribundum.

Over-expression of Arabidopsis thaliana AtNPR1 gene in rice enhances the rice resistance to bacterial blight and blast diseases

2016/1/15 2:42:12

The AtNPR1 gene is a key regulator of systemic acquired resistance(SAR)in Arabidopsis thaliana. Overexpression of AtNPR1 in A.thaliana enhanced resistance to both bacterial and fungal diseases. To investigate the effect of AtNPR1 over-expression in rice on its disease resistance, the gene was transformed as a restorer in Gui99 of Guangxi. PCR and DNA dot blot hybridization analyses revealed that 79 transgenic plants were obtained. Northern hybridization and RT-PCR analyses indicated that AtNPR1 gene was over-expressed in rice. The transgenic plants were challenged with Xanthomonas oryzae pv.oryzae(Xoo)and Magnaporthe grisea,the rice bacterial blight and fungal blast pathogens,respectively. They showed significantly enhanced resistance to Xoo and M.grisea.

Effects of ABA on biochemical parameters of normal and irregular leaves of Bougainvillea spectabilis

2016/1/15 2:42:12

The effects of ABA on some biochemical parameters of normal and irregular leaves of Bougainvillea spectabilis Willd were studied. Normal and irregular leaves of B.spectabilis were treated with ABA for 6 hours at different concentrations. Then the contents of chlorophyll, soluble sugar,soluble protein,free proline and SOD enzyme activity were measured. The results showed that the contents of chlorophyll,SOD activity,and free proline of irregular leaves and soluble sugar of normal leaves increased at first and then decreased and they reached maximum when treated with 100 μmol/L ABA. The soluble sugar of normal leaves,the protein content of normal and irregular leaves reached maximum when treated with 50 μmol/L ABA. This indicated that 50-100 μmol/L ABA could improve the resistance of B.spectabilis.

The protoplasts isolation and culture of Pogostemon cablin cv.Shipaiensis

2016/1/15 2:22:26

A protocol for protoplasts isolation and culture from the cell suspension of Polgostemon cablin cv.Shipaiensis were developed. The suspention cells grew 3-11 days after subculture was incubated in the enzyme solution consisting of 0.5%(w/v)pectolyase Y-23,0.2%(w/v)macerozyme R-10 and 0.8%(w/v)cellulase R-10 in 9%(w/v)mannitol buffered with 0.1% MES and 0.02% CaCl2,for 8 h,the yield was 1.65×106 protoplasts·mL^-1 PCV(Packed Cell Voloume)and the viability was above 86%. In thin layer culture of protoplasts,observed division rate was 13.5%.

Effects of temperature stress on germination and vitality of Papave somniferum seeds

LI Yan-Rong, CHANG Ying, WEI Yu-Jie, ZANG Guang-Peng — 2016/1/15 2:22:26
The effects of temperature stress on the germination and vitality of seeds of Papave somniferum L. were studied to preserve poppy germplasm resources and prevent seed aging. The results showed that poppy was high temperature sensitive and was able to germinate under low temperature. The favorable temperature for their germination was 10-20 ℃,and the optimal temperature was 18 ℃,the temperature lower than 5 ℃ and higher than 20 ℃ all could affect its germination. The temperature should be controlled within 60 ℃ and the heating time under 60 minutes when seeds pelleting. Under normal inventory conditions,with storage time increased,the poppy seed vigor dropped,and germination rate declined less than 50% as the storage time over 5 years,and was not suitable for breeding,so as the germplasm resources,it must breed again every 5 years.

Correlation between changes of photosynthetic characteristics of different sugarcane varieties and their cold tolerance under low temperature

2016/1/15 2:22:26
In order to explore the correlations between changes of photosynthetic characteristics and cold tolerance in different sugarcane varieties,7 newly bred sugarcane lines and 2 main cultivars from Sugarcane Research Institute,Guangxi Academy of Agricultural Sciences were employed to examine the changes of chilling injury index,chlorophyll content and photosynthetic perameters under low temperature stress at seedling stage. The chilling injury index was continually increased with the prolonging of low temperature treatment,but different sugarcane varieties showed different changes. The chlorophyll content in all the sugarcane varieties were continually decreased with the prolonging of low temperature treatment. There were significant differences in net photosynthetic rate and stomatal conductance between room temperature treatment and low-temperature treatment. The Fv/Fm,ΦPSⅡ,Fv'/Fm',qP and ETR were decreased obviously,but Fo,Fs and qNP were increased after the low temperature treatment. There were significant correlations between chlorophyll content,various photosynthetic perameters and chilling injury indexes,and the correlation coefficients between Fv/Fm,Fv'/Fm',ΦPSⅡ and chilling injury indexes were higher than 0.800,which suggested that Fv/Fm,Fv'/Fm',ΦPSⅡcould be used as the indexes of cold tolerance identification in sugarcane.

Four halophytes(or salt-tolerant species)and their accessory structures of leaf related to adversity tolerance in the desert of northern Xinjiang

2016/1/15 2:22:26
Paraffin sections,scanning electron microscope and temporary slide methods were employed to study the accessory structure related to adversity tolerance of four halophytes(or salt-tolerant)in the desert of northern Xinjiang,which included Karelinia capsia,Chenopodium glaucum,Salsola ikonnikovii and S.ferganica. The results indicated that:(1)the anatomical structure of the leaf and stem revealed:all the leaves were observed with the keratinized cuticle and sunk stomata,and a number of vascular bundles existed in the stems; many of crystals were presented in leaf and stem cells;(2)observation of the accessory structures of different leaves showed that large amount of salt bladders were discovered on epidermis of C.glaucum,which was consisted of 1-2 small stalk cell and a large bladder-like cell; there were short and hard trichomes and small papillary structure on the epidermis of S.ikonnikovii; there were large number of epidermal trichomes with nodes on the leaf of S.ferganica; K.capsia had multicellular salt glands with a black cap cell on the top and multicellular trichomes on the epidermis. Above structures and features reflected the diversity of adaptability of different plants to the salinity and drought environments,and which would provide some new experimental evidence in physiology aspect for the halophytes and xerophytes.

Search for proper dose of ⁶⁰Co-γ ray in Actinidia chinensis radiation breeding

2016/1/15 2:22:26
In this experiment red flesh Actinidia chinensis‘HongYang',yellow flesh A. chinensis‘GuiHai No.4' and green flesh A. chinensis ‘ChangGuo'branches were treated with ⁶⁰Co-γ ray in dose of 25 Gy,50 Gy,100 Gy and 150 Gy respectively and cutivated in randomized blocks design after treatment. The untreated branches used as control. Surivial rate and height of plant were examined. The result indicated that it was obviously that the inhibition of both the survival rate and the height of plant would be enhanced with increase of the radiation dose ranged in this experiment. Different varieties of A. chinensis quite different sensitivity to radiation. The medial lethal dose of ⁶⁰Co-γ radiation for these three varieties of A. chinensis branches were 50.6 Gy(Hong Yang)、64.4 Gy(GuiHai NO.4)、71.7 Gy(Chang Guo). The results would provide an important experimental basis for the radiation breeding of other varieties of A. chinensis.

Relationship between postanthesis carbon and nitrogen remobilization and grain growth of high quality rice under different water management

2015/12/16 11:40:11
Field experiments were conducted in a randomized complete block design with two factors with three water management，wettingdrying irrigation of water(WIW)，deficit irrigation of water(DIW)，submerged irrigation of water (SIW) and two indica high quality rice genotypes (Baguixiang and Guihuazhan) to investigate the relationship between postanthesis carbon and nitrogen remobilization and grain growth of high quality indica rice under different water management in 2007 and 2009. Results indicated that grain growth was closely correlated with carbon and nitrogen remobilization under three different water management conditions.（1）dry matter contribution rate of culm and sheath and leaf blade to grain were from 16.86% to 25.68%. Initial grain filling potential was significant positive correlated with dry matter remobilization rate and efficiency of culm and sheath；maximum grain filling rate ，active filling period and active grain filling duration were significant positive correlated with leaf dry matter remobilization rate and efficiency after flowering and significant negative correlated with clum and sheath；（2）the contribution rate of the remobilization of carbon assimilation of culm and sheath to grain yield and starch yield was WIW＞DIW＞SIW，however，the contribution rate of the remobilization of carbon assimilation in leaves to grain yield and starch yield was SIW＞DIW＞WIW. The soluble sugars accumulation of stem and leaf reduced along with the increase in grain amylose content and accumulation increased. The rapid decline period (after flowering 3-12 d) of soluble sugar accumulation in leaves was synchronized with the rapid increase amylose content and accumulation period (after flowering 6-12 d)；（3）nitrogen contribution rate of culm and sheath and leaf blade to grain were 44.05% to 11766%. The contribution rate of leaf total nitrogen remobilization to grain nitrogen accumulation was greater than those of culm and sheath；the contribution rate of nitrogen assimilation of stem and sheath，and leaf to grain nitrogen assimilation was SIW＞DIW＞WIW.

Comparison of four alfalfa varieties’ several physiological items change in reagents of the acid， alkali and salt

2015/12/16 11:40:12
The comparison test was set up to know the four foreign alfalfa varieties’physiology response under adversity conditions and confer their antiintimidating traits according to their sensitivity to the adversity factors，and convenient for adjusting measures to local conditions and exerting its most productivity. Using vitriol and sodium hydroxide and lixivium from soil with different pH value reagents，the pH values were from 3 to 12，and there were ten steps altogether；when the pH value was 7 with different thickness of salt reagents，they were respectively 0.1%，0.2%，03%，0.4%，0.5% and 0.6%；once more，when the pH values were 8，9，10，11，12 respectively，with 0.2%，0.3%，0.4%，0.5% and 0.6% salt thickness accordingly. Then four alfalfa varieties were planted，their physiology items were measured and to their sensitivity was analysed. The results showed that the character of acidresistant:Gonden Empress＞Grangeur＞Algonquin＞Victoria；the character of alkaliresistant:Grangeur＞Algonquin＞Gonden Empress＞Victoria；the character of saltresistant:Grangeur＞Gonden Empress＞Algonquin＞Victoria；the character of salt and alkaliresistant: Grangeur=Algonquin＞Gonden Empress＞Victoria. Therefore，four alfalfa varieties should be plant them in different soil，according to their character of the antiintimidating.

Probing the cause of the low germination rate of Merremia boisiana seed

2015/12/16 11:40:12
To find out the reasons for its indoor low germination rate，the permeability of seed coat，seed anatomical structure，seed vigor and seed endogenesis inhibitors were examined. The results showed that the permeability of seed coat was poor. Chipped seeds reached the highest value of water absorption in less than 38 h as the intact seeds. The proportion of empty seeds was as high as 30%. Seed vigor was low and averaged at 35%. The germinability，shoot and root growth of cabbage seeds were significantly inhibited by methanol extracts of M.boisiana seeds. When methanol extracts of seeds reached a concentration of 25 mg/mL，germination and growth of cabbage seeds were seriously inhibited，suggesting rather high content of endogenesis inhibitors in M.boisiana seeds. Low germination rate of M.boisiana seeds indicated that seed's generation of a new plant was not the main reason for the outburst overspread disaster in the recent years, and the key factors should be further researched.

Effects of light and temperature on seed germination of Eupatorium odoratum

2015/12/16 11:40:12
The effects of different light and changing temperature conditions on seed germination of Eupatorium odoratum  were studied by simulating environmental conditions in climatic chambers. The results was as followed:the seed of E.odoratum could germinate from 15℃/10℃ to 40℃/35℃ under light，and the optimum changing temperature was 30℃/25℃. Under this changing temperature，the germination rate was 47.5%；at 15℃/10℃，the seed of E.odoratum couldn't germinate under dakness；from 20℃/15℃ to 35℃/30℃ temperature conditions，the higher temperature，the seed germination peak appeared earlier；from 15℃/10℃ to 30℃/25℃ temperature conditions，the seed germination rate of E.odoratum increased with increasing temperature，and higher than 30℃/25℃，the germination rate reduced. Appropriate light was more conducive to seed germination of E.odoratum. E.odoratum became invasive species and proliferated speedily could be closely related with seed germination adaptability to light and temperature.

Study on in vitro propagation and tube plant vitrification of Dianthus chinensis

2015/12/16 11:40:12
Effects of medium ingredients on seeds germination，buds inducement proliferation，tube plant vitrification and rooting were studied in order to establish efficient rapid propagation technology system of Dianthus chinensis cultivar Diana F1 White. The results showed that proper seeds germination medium was 1/2 MS，and seed germination ratio was 83.33%. Height of aseptic tube plant reached to 6.99 cm，and leaf number per plant was 26.7 on 30th day after sowing in medium. Max.vitrification plant ratio was 83.33% during bud inducement period for tube plant of Diana F1 White. The optimal medium for buds inducement was MS+6BA 3.0 mg/L+NAA 0.3 mg/L+agar 8.0 g/L+sugar 8.0 g/L after light intensity was promoted to 2 000 lx. Bud inducement ratio and buds number per explant were 90% and 7.2 respectively，and the ratio of vitrification tube plants decreased to 3.33% after 30 days’culture. The optimal rooting medium was 1/2 MS+1.0 mg/L IBA+agar 8 g/L+sugar 40 g/L，and rooting percent，roots number and average root length was 100%，24.7 and 4.7 cm respectively after 30 days’ culture. Survival rate of tube plant was more than 95% after they were transplanted in disinfected humus. The above results would provide scientific base for micro propagation of D.chinensis cultivar Diana F1 White.

Comparison and analysis of SCoT and ISSR markers for genetic diversity of longan

2015/12/16 11:40:12
The diversity and genetic relationship among and within thirtysix varieties of longan and one Dimocarpus confinis from China，Vietnam and Thailand were analyzed using SCoT（Start Codon Targeted） and ISSR(Inter Simple Sequence Repeat) markers. The results showed 127 strips were amplified by 12 SCoT primers while 117 strips by 15 ISSR primers. The average strips amplified by SCoT and ISSR primers were 10.58 and 7.8，respectively. The genetic relationships were analyzed using unweighted pairgroup method of arithmetic average cluster analysis(UPGMA)and the genetic Jaccard similarity coefficients were calculated. The cluster analysis results showed that the thirtyseven materials could be clustered into 6 groups at the similarity coefficient level of 0.672 and 0.685. Comparison and mixture analysis of the two molecular markers demonstrated that both SCoT and ISSR were efficient approaches for genetic diversity analysis of longan germplasm. These results would provide theoretical basis for longan germplasm storage and utilization in future.

Preliminary study on genetic diversity of Magnolia sargentiana (Magnoliaceae) through SNP marker

WANG JiaYuan1， WU ChuanFang2， TANG Ya1* — 2015/12/16 11:40:12
Magnolia sargentiana is a species endemic to China and is listed on the Red List of vulnerable species. It is only found in limited localities in Sichuan and Yunnan provinces. Using samples collected from the Mamize Provincial Nature Reserve and the Meigu Dafengding National Nature Reserve in the southern Sichuan Province，its genetic diversity was investigated with single nucleotide polymorphism (SNP) marker，based on PCR and sequencing technique. In the four sequences (510 bp lengths) that amplified with specific primers，a SNP locus was found on every 73 bp sequence on average，which suggested that the different M.sargentiana populations in the two nature reserves had a high level of genetic diversity；the similarity among the amplified sequences reached to 97%，and 36% of the amplified sequences was similar to the sequence used for making primers，which indicated that the sequence we selected was suitable for genetic research of M.sargentiana. The results would provide useful information for further conservation research and protection policy making on M.sargentiana or other protected species.

Cloning and sequence analysis of full length of mulberry (Morus alba) actin gene

KONG WeiQing， YANG JinHong — 2015/12/16 11:39:57
As a standard reference model of gene expression and regulation，actins play important roles in plants. In this study，the full length 1 612 bp of actin gene from mulberry was cloned by means of chromosome walking. Its CDS was 1 312 bp in length，encoding 377 amino acid residues. Homology analysis of the deduced amino acids showed above 90% identity with the actins from rice and grape，et al. The number and site of introns was conservative with rice and grape，too. Cluster analysis of actins from 24 species indicated that these genes were divided into two subgroups of Class I and Class Ⅱ.

Cloning and identification of APX gene from Isatis indigotica

SUN XiaoDong*， LI AiLing， HAN LiMin — 2015/12/16 11:39:57
To construct the recombinant eukaryotic expression plasmid of pTZ57R/T/APX. APX gene was amplified from the genomic DNA of Isatis indigotica by polymerase chain reaction(PCR). APX gene was then inserted into pTZ57R/T eukaryote expression vector. The positive colonies were screened and identified by PCR 〖JP3〗and sequencing. The specific gene APX which was about 1 257 bp，was successfully amplified and pTZ57R/TAPX were constructed. The DNA sequence of APX gene was as the same as the APX gene nucleotide sequences published in Genebank. pTZ57R/TAPX recombinant was successfully constructed. And the construction provides the basis for the further study.

Alternative splicing of flowering regulatory gene LFY in Arabidopsis thaliana

YU LiXia， YAO YouLin， WU XiaoLu， TANG XiaoQian， YAN Bo* — 2015/12/16 11:39:57
To study the expression of flowering regulatory gene LFY gene in Arabidopsis thaliana，we used reverse transcriptionpolymerase chain reaction and isolated three alternative splicing(AS)fragments named as LFY1239，LFY1263，and LFY1275 respectively. Sequence analysis confirmed that fragment LFY1263 contained an open reading frame of 1 263 bp，and was identical to the previously reported and predicted fragment，while LFY1239 lacked 36bp at the 3′ end of the first exon，and LFY1275 had an additional 12bp derived from the 3′ end of the first intron. Expression analysis showed that LFY1239 was only detected in rosette during the vegetative stage，while LFY1263 and LFY1275 presented in both floral organs and rosette during both vegetative and flowering stages. Furthermore，LFY1263 appeared to be the most abundant transcript. The expression ratio of LFY1275 to LFY1263 was higher in floral organs than in rosette leaves，which suggested that such ratio might be associated with the flowering regulation.

Effects of lanthanum on the active oxygenscavenging enzyme activities and ultrastructure in Potamogeton crispus leaves

HU JinZhao — 2015/12/16 11:39:57
The ultrastructural and physiological effects of lanthanum on Potamogeton crispus were studied in present hydroponic experiment. It was found that the foliage grew better in LaCl3·8H2O than in the medium without LaCl3·8H2O. The chlorophyll content，average auto-fluorescent intensity，nitric acid reducase(NRase)activity，peroxidase(POD)activity，catalase(CAT)activity and soluble protein content increased when the plants were treated with lower La3+ concentration，whereas superoxide dismutase(SOD) activity，membrane permeability，O〖ST4.〗2〖HT〗. generation rate and malondialdehyde(MDA) content decreased. The TEM results provided an ultrastructural evidence for the effect of La3+ on P.crispus. The optimum La3+ concentrations which promote P.crispus growth were suggested to range from 10 to 15 mg·L1.

Heterosis analysis of F2 kernels nutrition in shsu double recessive sweet corn

CHENG XinXin， LIU Zheng — 2015/12/16 11:39:57
Heterosis is an universal phenomenon of genetics. The heterosis of sweet corn in different characters were different，and the genetic characteristics and heterosis of F2 kernels of 10 cross combinations with 5 shsu inbred lines were analysised according to the P(P-1)/2. The results showed that the 4 quality traits of F2 kernels were difficult to achieve the desired effect at the same time. The total soluble sugar content，cane sugar content，starch content and soluble protein content in kernels were affected by parents which were basically influenced by a low parental content. In general，the parents with high content of total soluble sugar were used in order to get higher F2 kernels and cane sugar content and starch content were influenced by mailparents.

Influence of colchicine on radicle growth of Vicia faba and the polyploid inductive effect

YAN QiuJie， YANG Qiong — 2015/12/16 11:39:57
In order to test the optimum induction combination(time and concentration)，and analyse the inductive effects of colchicine induced the radicle polyploid in Vicia faba. The radicle cells were treated with the colchicine of 〖JP3〗five concentrations (0.025%，0.050%，0.100%，0.150%，0.200%，respectively) combining various culture time (12，24，48，60 h，respectively). And the radicle cell were treated with distilled water simultaneously，which were taken as the control. The 〖JP2〗optimum induction combination induced by colchicine was confirmed by significant difference of the root tip inductivity and the radicle expanding ratio. And the chromosome number of radicle cells and stoma number of seedling were detected to identify the induction effect. The radicle amount and seeding height were analized subsequently. The optimum inductive condition was 48 h combining colchicine concentration of 0.100%. There were significant differences in stoma number between inducted time and inducted concentration，however，the radicle number and height of V.faba seedling showed significant difference in inductive concentration but no significant difference in culture time.

Preclinical pharmacokinetics of novel soybean iosflavone sulfonate

PENG You1，2*， TAO ChunYuan1， DENG ZeYuan2 — 2015/12/16 11:39:58
The oral bioavailability of genistein (GE) in its benzensulfonate prodrug was studied in search for its novel prodrug. The plasmas were collected at different points of time after the intragastric or intravenous administration of genistein benzensulfonate (GBS) 40 mg/kg to rats. The GBS and GE contents in plasma were determined by HPLC. The compartment model was fitted and pharma cokinetic parameters were calculated by DAS 2.1.1.The results indicated that the dynamic processes of GE were consistent with two compartment model after intragastric administration of GBS prodrug to rats. The relative oral bioavailability of GE in prodrug GBS was 198.6%. In conclusion，the above results demonstrated that the oral bioavailability of GE in prodrug had been improved remarkably.

Antimicrobial activities of 26 Yunnan plants extracts against clinical multidrug resistant pathogens in vitro

YANG CuiXian1，2， ZUO GuoYing1*， HAN Jun2， WANG GenChun1 — 2015/12/16 11:39:58
To determine the in vitro antimicrobial activities of 80% ethanol extracts from 26 plants originated in southeast of Yunnan. The ethanol extracts of the collected plant samples were subjected to screen of the antibacterial and antifungal activity by the agar hole diffusion test. The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)or minimum fungicidal concentration(MFC)were determined by serial dilution with a standard broth microdilution method. The results showed that 9 of the 26 extracts were active with different potency against strains of Staphylococcus aureus，Escherichia coli，Pseudomonas aeruginosa and Candida albicans. The most active extracts against Staphylococcus aureus and methicillinresistant S.aureus(MRSA)were from Rhodoleia parvipetala Tong，Diplopanax stachyanthus Hand.Mazz，Sarcosperma kachinense Exell var.simondii Lam.et Royen，Coelogyne leucantha W.W.Smith and Sunipia bicolor. Their MICs/MBCs(mg/L)were 512/2048＞2048，512/2048＞2048，256512/2048＞2048，5121024/1024＞2048 and 5121024/＞2048，respectively. Sladenia celastrifolia Kurz showed strong activity against P.aeruginosa and C.albicans，and the MICs/MBCs(MFCs)were 5122048/＞2048 and 2048/＞2048 mg/L，respectively. All these plants showed weak inhibition against Escherichia coli.

Effects of Ceratocystis fimbriata on phenolics content， PPO and PAL activity in sweet potato

WANG JingJing1， LIU MeiYan1， XIE YiPing2， SUN HouJun2， ZHANG Jian1* — 2015/12/16 11:39:58
Two varieties of sweet potato including high resistanceNanjing92 and high susceptibleYantai252 to Ceratocystis fimbriata were used for detection the content of total phenol，chlorogenic acid，flavonoid and the activity of PPO and PAL in sweet potato leaves. The results showed that flavonoid content，chlorogenic acid content and PAL activity of Nanjing92 were higher than that of Yantai252 and even reached significant or extremely significant difference in normal condition，which could be regarded as physiological indices of identifying and selecting resistant variety. It was also found that total phenol content and PPO activity of two varieties were not significant different in normal condition，however，when infected by C.fimbriata，total phenol content and PPO activity of Nanjing92 increased sharply and significant or extremely significant higher than that of Yantai252 within 2th to 8th day. The total phenol content and PPO activity raised speedily and maintained long period which could increase the resistance ability to C.fimbriata of sweet potato.

Identify and genetic analysis on the cryotolerance of overwintering cotton in Nanning， Guangxi

2015/12/16 11:39:43
The overwintering cryotolerance of cotton materials derived from different germplasm and their hybrids were identified in Nanning，to provide the basic materials for perennial cotton breeding in cryotolerance. Based on the selection of the optimal stress temperature and time，5 physiological and biochemical indexes(relative conductivity，soluble sugar，soluble protein，MDA and proline)of cryotolerance and overwintering survivorship in open field were used to identify comprehensively on the overwintering cryotolerance of 63 materials，then they were clustered into 4 categories:23 materials in cryotolerant type，19 materials in middle cryotolerant type，9 materials in middle cryosensitive type and 12 materials in cryosensitive type，accounting for 36.51%、30.16%、14.29% and 19.05% respectively. All materials in cryotolerant type were relative with germplasm of perennial island cotton，which indicated that the cryotolerance of overwintering in perennial island cotton was very useful for breeding. Cytoplasm effect of overwintering survival wasn’t significant in testing cotton materials，the overwintering survival of hybrids were in regression trend of mean type in ancestors.

Optimization of genetic transformation system of potato and introduction of maize starch branching enzyme gene SBEⅡb into potato

FAN YaLi1， RUAN Ying2*， LIU ChunLin2 — 2015/12/16 11:39:43
The highly effective potato transformation systems were established and optimized using tube potato plantlets as transformation materials in this study. The maize starch branching enzyme 〖JP〗gene(SBEⅡb)was transformed into potato using Agrobacterium tumefaciensmediated transformation method. 762 stem sections were infected by Agrobateriummediated tumefacien method. 4 transgenic plants were obtained by PCR measurement，implying that SBE Ⅱb 〖JP2〗gene had been integrated into potato genome. It was also found that GUS gene could normally be expressed in stem sections and potato microtubers of the trasgenetic potato plants.

CP gene cloning and sequence analysis of Liu’an isolates of Leek yellow stripe virus， Onion yellow dwarf virus and Shallot yellow stripe virus

WEI ChuanBao1，2*， HUA JunYa1， YANG Yu1， WEI YangYang1， SHI LiLi1 — 2015/12/16 11:39:43
Specific primers were designed to amplify CP genes of Leek yellow stripe virus(LYSV)，Onion yellow dwarf virus(OYDV)and Shallot yellow stripe virus(SYSV)Liu’an isolates. Then the CP genes were cloned into pGEMT vectors and were sequenced. Sequence analysis indicated that CP genes of LYSV，OYDV and SYSV Liu’an isolates consists of 864，771 and 774 nucleotide acids respectively. Multiple aligments showed that CP gene of LYSV Liu’an isolate shared 76.12%-84.31% nucleotide acids identities with 68 LYSV CP genes reported on Genbank，CP gene of OYDV Liu’an isolate shared 81.06%-90.40% nucleotide acids identities with 86 OYDV CP genes reported on Genbank and CP gene of SYSV Liu’an isolate shared 88.63%-94.32% nucleotide acids identities with 11 SYSV CP genes reported on Genbank. These results indicated that the divergence in LYSV CP among different isolates was high，whilst divergence of OYDV CP was moderate in different isolates. All the results indicated that the variability of LYSV CP gene among different isolates was high，OYDV CP gene among different isolates was in the middle and SYSV CP gene among different isolates was low. Phylogenetic analysis showed that LYSV CP had little relationship with OYDV CP and SYSV CP and OYDV had closer relationship with SYSV than LYSV.

Study on technology of aseptic sowing and rapid propagation of Demdrobium officinale

2015/12/16 11:39:43
Seeds of Demdrobium officinale could be used as explants to rapid propagate by the way of seed→protocorm→the whole plant→transplant，and medium in each culture stage and some other factors were studied comparatively. The results showed that all seeds of D.officinale which were 60-180 d，especially 150-180 d after pollination could germinate，with a germination rate of 87.2%-94.4%. The suitable medium for seed germination was MS+6BA 1.0 mg/L+NAA 0.1 mg/L+potato 200 g/L+active carbon 1.0 g/L. The best medium for the formation and multiplication of protocorm was MS+6BA 1.5 mg/L+NAA 0.1 mg/L+banana mud 100 g/L+active carbon 1.0 g/L，the propagation coefficient was about 20 times/50 d. And the protocorm differentiated on MS+6BA 1.0 mg/L+NAA 01 mg/L+potato 200 g/L+active carbon 1.0 g/L medium，which could have some multiplication in the meantime. After being cultured on MS+6BA 0.5 mg/L+NAA 0.2 mg/L+banana mud 100 g/L+active carbon 1.0 g/L for a generation，the vigorous plantlets were transferred to rooting medium which was 1/2MS +NAA 0.8 mg/L+mineral salt A 02-0.5 mg/L +banana mud 100 g/L+active carbon 1.0 g/L and were cultured for 50-70 d，with 100% rooting rate. Mineral salt A could effectively control the formation of callus or protocorm，and obviously improve the quality and quantity of root regeneration plants. The best time for the transplantation of root regeneration plants was March  May and September  October in Guilin area. The plantlets were transplanted into rotten pine bark handled with high temperature in seedbed of greenhouse resulted in more than 90%survival rates.

Comparative study on several physiological characters among the reintroduced population of Cycas debaoensis

LI ZhengWen1， CHEN LiLi1，2， LI ZhiGang1， LI SuLi1， LI Nan2* — 2015/12/16 11:39:43
Comparative study on the activity of superoxide dismutase(SOD)，catalase(CAT)，peroxidase(POD)，malonal dehyde(MDA)as well as the content of proline，soluble sugar and soluble protein in leaves of the seedlings of Cycas debaoensis in Huanglianshan Nature Reserve of Guangxi was carried out in the present paper. Results showed that the activity of SOD and CAT was increased in the whole development stage，while CAT activity decreased at earlier stage and then increased；MDA activity declined；proline showed a slow increasing tedency and the soluble sugar and soluble protein continuously accumulated. According to a comprehensive analysis，seedlings of C.debaoensis could be divided into two classes，plants with five leaves，which had a relatively higher environment adaptation；plants with three or four leaves merged into one class，which had a lower adaptability.

Comparative study of physiological indexes of Lilium formolongi under different photoperiod conditions

LIU Wei1， LIU JiuDong2，3， ZHOU HouGao4* — 2015/12/16 11:39:43
Three physiological indexes of Lilium formolongi under long day and short day conditions were studied. The results were as follows：physiological activities of the upper and middle part leaves had close relation with growth and development state of plant. For the upper and middle part leaves，soluble sugar and starch indexes decreased to a lower level，while total nitrogen index increased to a higher level at initial stage of bud morphological differentiation. Moreover，each C/N index of three parts leaves in reproductive growth state was higher than each C/N index of three parts leaves in vegetative growth state.

Response of cell membrane and lightresponse curve parameters of Spartina anglica to NaCl stress

SHI GuiYu1，2， KANG Hao1， YI LiNa1， LI MingXia1 — 2015/12/16 11:39:25
This research covers the study of lightresponse curve parameters，membrane permeability and malondialdehyde contents of Spartina anglica in NaCl salt stress. The results indicated that there was no significant change in membrane permeability or malondialdehyde contents at the salt concentrations no higher than 300 mmol•L1. In the same condition，the relative high Amax(＞30 μmol•m2•s1)，high apparent quantum yield(＞0.05 mol•mol1Photons)，relative low dark respiration rate(＜1.5 μmol CO2•m2•s1) as well as light compensation point(＜20 μmol•m2•s1) could be prerequisites for organic matter accumulation，competition，establishing populations and pervasion. However，at the salt concentration higher than 500 mmol•L1，the member permeability and malondialdehyde contents increased obviously，Amaxand apparent quantum yield decreased significantly, and dark respiration rate and light compensation point increased slightly. All these changes showed that cell membrane and enzyme of photosynthesis of S.anglica was damaged，so their growth was inhibited. In addition，relative lower photosynthetic rate and significantly lower transpiration rate of S.anglica under salt stress provided the conditions for their existence and growth by higher water use effective and remission of persecution of osmotic potential to cell membrane.

Moisture gradient affects the growth and physiological characteristics of Houttuynia cordata

WANG XiaoPeng1，3*， YE MeiRong1， YANG AnZhong2， SUN Zhen1， ZHANG Jian1 — 2015/12/16 11:39:25
This study aims to determine effects of moisture gradient on the growth and physiological characteristics of Houttuynia cordata for developing an ecological cultivation pattern. Eight kinds of water treatments were conducted to simulate the dynamic change of soil water moisture in the natural habitat of forest. The results showed that biomass was significantly reduced by decrease or increase of moisture as compared to the control；plant height and LAI were inhibited under the condition of heavy drought，moderate drought and waterlogging；heavy drought significantly elevated the root to shoot ratio；decrease or increase of moisture all increased the content of MDA except light moisture group；compared with the control，the activity of SOD and CAT，as well as content of MDA were significantly increased under the condition of drought；the activity of CAT，POD，SOD and the content of MDA of moisture group were respectively lower than those of drought group except for content of proline. In particular，the activity of CAT，POD and SOD，the content of MDA and proline of waterlogging group were generally lower than those of others. H.cordata had been stressed by the condition of moderate drought or moderate moisture. In summary，light drought and light moist conditions were favorable for the cultivation of H.cordata.

Molecular Cloning and expression analysis of 3hydroxy3methylglutarylCoA Reductase Gene from Eucalyptus urophylla×E.grandis

JIANG Chun1， ZHANG HuaLing2， PENG Jiang2* — 2015/12/16 11:39:25
The gene encoding 3hydroxy3methylglutarylCoA reductase (HMGR)was cloned by Rapid Amplification of cDNA Ends (RACE)method，the fulllength EuHMGR cDNA was 1955 bp，containing a 1 560 bp open reading frame，encoding a peptide of 519 amino acids. Bioinformatics analysis indicated EuHMGR contained two HMGCoA binding motifs and two NADP(H)binding motifs；and it had a 3D structure with“V”homologybased on modeling，containing Ndomain，Sdomain and Ldomain. There was a 319 bp intron in EuHMGR gene genomic DNA sequence compared with the cDNA sequence. Tissue expression profile analysis by RealTime quantitative PCR indicated EuHMGR expression was the highest in stem，followed by leaf 〖JP3〗and no expression in root. The paper would supply some information to the function analysis and genetic transformation of EuHMGR from Eucalyptus urophylla×E.grandis.

Research on callus induction and cell suspension culture of Atractylodes lancea

TAO JinHua1， PU XueLian1， JIANG Shu2 — 2015/12/16 11:39:25
Effect of different plant growth regulator on petiole，leaf and rhizome callus induction was studied by orthogonal design in this paper. It showed that petiole，leaf and rhizome callus induction rate were 99.0%、83.5% and 71.5% in their optimum culture conditions respectively. And Petiole was the most easy to form callus. There was a significant promotion effect of 2，4D on the callus induction. The optimal composition for petiole callus induction was 0.4 mg•L1NAA、4.0 mg•L1 2，4D and 0.4 mg•L1 KT；and induction rate reached 99.0% when petiole being cultured for 20 d. In addition，when cell from petiole callus was cultured for 18 d，the maximum yields of cell biomass，polysaccharide and atractylodin were 9.07 g•L1、15.68 mg•L1 and 19.62 ug•L1respectively.

Physiological and biochemical changes in lateral root mutant of rice

ZHANG Xia1， HUANG Bin1， LI Yang1， HAO ZaiBin1，2* — 2015/12/16 11:39:25
The activities of super oxide dismutase(SOD)，peroxidase(POD)，catalase(CAT)and succinate dehydrogenase(SDH)，the content of chlorophyll，malondialdehyde(MDA)，and proline(Pro)，and the electrophoresis of soluble protein were compared among no lateral root mutant(RM109)，original varieties(Oochikara)and their hybrids(F1). The results showed that SOD activity of Oochikara was stronger than RM109 by 50%，F1 was 72% of Oochikara；the POD activity of RM109 was higher than Oochikara by 125%，F1 was lower than Oochikara by 3%；the CAT activity of RM109 was 56% of Oochikara，F1 was only 11% of Oochikara；the SDH activity of RM109 was 60% of Oochikara. All above differences were up to be very significant. All of the revealed characteristics changes in physiology and biochemics might be caused by the gene mutation.

Application of ESIMS combined with 1H NMR in analyzing seed phospolipids of six species of Michelia

TANG AnJun1， Naomichi BABA2 — 2015/12/16 11:39:25
To date，there was no example to authenticate seed phospholipids based on their fingerprinting. By means of electrospray ionization mass spectrometry(ESI MS)and proton nuclear magnetic resonance(1H NMR)spectroscopy，phospholipids fraction extracted from seeds of six species from the genus Michelia were detected. It was firstly found that distinct difference in spectral fingerprinting region between m/z 895910 in ESI MS and 5.30-5.40 mg/L in 1H NMR among these six species. Thus，it suggested that the spectral differences shown by ESI MS and 1H NMR among seeds of these six species could be applied to identify them. And in a wider sense，for analyzing seed phospholipids of other plant species，a combination of ESI MS AND NMR was an effective tool.

Study on dynamic accumulation of glycosides in Picria felterrae

NING DeSheng， FANG Hong， LIANG XiaoYan*， CHEN HaiShan — 2015/12/16 11:39:25
The content of glycosides of P.felterrae in different growth period were detected with HPLC to reveal its dynamic accumulation. The correlation analysis of glycosides and the growth situation was conducted throught comparing the chromatograms and peak areas of different growth period. The results showed that the variation trend of content of glycosides，picfeltarraenin IA and Ptarraenin IB was nearly consistent. The content of picfeltarraenin IA and Ptarraenin IB both reached to the maximum when the plant grew up to 75 days，then subsequently decreased slowly，but increased again in early fruit stage(namely 120d)when the biomass reached to the maximum. Therefore，it was suggested that the best collection stage for P.felterrae was the early fruit stage(120 d).

Studies on isolation、purification、structural identification and its antitumor activity of polysaccharides from Momordica grosvenori swingle’s root

YAN XiaoJie1，2， LU FengLai1， CHEN HuanYing1，2， LI DianPeng1* — 2015/12/16 11:39:25
Polysaccharide(CPS)of Momordica grosvenori swingle’s roots was extracted by water extractingalcohol precipitating method，and then was pufified by DEAE52 cellulose. Their compositions were analyzed by paper chromatography and high performance liquid chromatography(HPLC). And the effect of CPS on H22 subcutaneous planting of mice was studied. Results:CPS was isolated into five fractionates by DEAE52 cellulose column chromatography，its simple sugars composition included glucose，arabillose and xylose. Compared with model group，each dose group had no significant functions in suppressing tumor growth(inhibition rate＜50%)；Besides，Compared with model group，each dose group had improved influence on the index of thymus(P＜0.01)，but had reduced influence on the index of spleen(P＜001).

Physiological response of Spartina alterniflora seedlings to cadmium stress

2016/1/20 18:33:54
The physiological and biochemical response of Spartina alterniflora seedlings to different concentration of Cd investigated by cultivating the material in the simulated seawater and sand. The results showed that as the concentration of Cd increasing,the biomass of S. alterniflora leaves and roots was decreasing,while the membrane permeability,malondialdehyde(MDA),the enzymatic activity of SOD and POD tended to reduce,so S. alterniflora could make the best of its self-resistance. With the increase of the concentration of cadmium,photosynthesis characteristic of S. alterniflora changed greatly. The net photosynthetic rate,intercellular CO₂ concentration(Ci)and stomatal conductance(Gs)dropped and reduced.

Shoot growth controlled by gorwth regulator in longan

2016/1/20 18:33:54
The effects of shoot growth controlled by growth regulator spraying on flower bud differentiation(red point appearing)and flower inflorescence spindle elongation 6-9 cm and leaf expansion was studied in order to regulate shoot growth safely and efficiently. The results indicated that foliar-spraying 200 mg/L ethephon and 150 mg/L paclobutrazol,or soil application 4 grams per plant paclobutrazol had the best effect on preventing shooting inflorescence from branch leaves. Shooting of inflorescence was singificantly controlled by 300 mg/L ethephon with siphon infusion.

Influence of shading on net photosynthetic rate and quality of Siraitia grosvenorii cultivated in south Guangxi

2016/1/20 18:33:54
Different shading treatments(shading rate 0,30%,50% and 70%)were used to study the influence on net photosynthetic rate and quality of Siraitia grosvenorii at the late growth stage through the field conditions. The diurnal variation of net photosynthetic rate varied from a bimodal curve under control group(shading rate 0)to a unimodal one under shading condition and curve peak value appeared at 12:00 noon. Curve peak value,the content of mogroside Ⅴ and dried fruit weight under the treatment of 50% shading rate were higher than those under the treatments of other shading rates.The results showed that at the late growth stage of S. grosvenorii cultivated in south Guangxi,the treatment of 50% shading rate increased net photosynthetic rate,and it was good for dried fruit weight and improving the content of mogroside Ⅴ.

Applications studies of the modified exponential model on photosynthesis—light response and CO₂ response curves of Mirabilis jalapa

2016/1/20 18:33:54
To study changes of photosynthetic efficiency parameters and explore the applicable model,five models were used to describe photosynthesis-light response and CO₂ response curves of Mirabilis jalapa,respectively. The results were as follows:(1)The light response and CO₂ response curves fitted R² of modified exponential model were both 0.999,the light saturation point and the maximum net photosynthetic rate were 797.299 μmol·m^-2·s^-1 and 7.879 μmol CO₂·m^-2·s^-1,respectively; the CO₂ saturation point and the photosynthetic capacity were 1 264.447 μmol·mol^-1 and 16.783 μmol CO₂·m^-2·s^-1 respectively,which most proximally fitted the measured values. The modified exponential model was superior to nonrectangular hyperbola model,rectangular hyperbola model,modified rectangular hyperbola model and exponetial model;(2)The mean square error(MSE)and mean absolute error(MAE)of fitted and predicted values in the modified exponential model were much lower than those in other models. In conclusion,the modified exponential model was optimal model for describing the photosynthesis-light response and CO₂ response curves of M. jalapa,which could provide a reference application in physio-and ecology of M. jalapa.

Effects of trace elements on chlorophyll fluorescence parameters of clonal eucalypt seedling

2016/1/20 18:33:54
The effects of level and form of trace elements on chlorophyll and its fluorescence in seedling leaves from 3 eucalypt clones were studied by way of sand culture. The results showed that the chlorophyll content and fluorescence parameters were influenced by factors significantly. The eucalypt clone was an important effective factor and had high significantly influence on all traits. In the factors of trace elements,there existed high significant differences in Fe(B)to Fo,Fm,Fv/Fo and Fv/Fm,Zn(C)to Fo and Fv/Fm,Mn(D)to chlorophyll content(SPAD value),Mo(F)to chlorophyll content and Fo,B(G)to chlorophyll content,Fo,Fv/Fo and Fv/Fm,respectively. There were also significant relationships between Fm and Fo,between chlorophyll content and Fo,Fm,Fv/Fm and Fv/Fo.With the increase of Fe,Zn,Cu,Mo,B solution nutrition,Fo and Fv/Fm increased,but Fv/Fm decreased under the higher concentration. Compared with chlorophyll fluorescence,U₆ and DH_32-29 had higher Fo,Fm value and lower Fv/Fm,Fv/Fo value,but DH_32-29was more susceptible to abundance and deficiency of trace elements in soil. The results also showed that applying chlorophyll fluorescence would be a good method to evaluate the health of eucalypt species in trace element nutrient.

Applications studies of the modified exponential model on photosynthesis—light response and CO₂ response curves of Mirabilis jalapa

2016/1/20 18:33:54
To study changes of photosynthetic efficiency parameters and explore the applicable model,five models were used to describe photosynthesis-light response and CO₂ response curves of Mirabilis jalapa,respectively. The results were as follows:(1)The light response and CO₂ response curves fitted R² of modified exponential model were both 0.999,the light saturation point and the maximum net photosynthetic rate were 797.299 μmol·m^-2·s^-1 and 7.879 μmol CO₂·m^-2·s^-1,respectively; the CO₂ saturation point and the photosynthetic capacity were 1 264.447 μmol·mol^-1 and 16.783 μmol CO₂·m^-2·s^-1 respectively,which most proximally fitted the measured values. The modified exponential model was superior to nonrectangular hyperbola model,rectangular hyperbola model,modified rectangular hyperbola model and exponetial model;(2)The mean square error(MSE)and mean absolute error(MAE)of fitted and predicted values in the modified exponential model were much lower than those in other models. In conclusion,the modified exponential model was optimal model for describing the photosynthesis-light response and CO₂ response curves of M. jalapa,which could provide a reference application in physio-and ecology of M. jalapa.

Cloning and bioinformatic analysis of SAMS gene in a mangrove tree Sonneratia alba

2016/1/20 18:33:54
Sonneratia alba is one of the most salt-tolerant mangroves. S-adenosylmethionine synthetase(SAMS)which was in response to abiotic stresses played an important role in salt tolerance. In order to study the biological mechanism of S. alba for adapting stress,the coding region of SaSAMS gene which codes a key enzyme in the S-adenosylmethionine(SAM)biosynthetic pathway was isolated. Based on the annotation of root transcriptome of S. alba,primers was designed and cDNA of SaSAMS gene was amplified by PCR. The complete coding sequence of SaSAMS gene was 1 182 bp and it encoded a protein of 393 amino acids. Homology comparison and evolutionary tree analysis showed that the SAMS amino acid sequences were relatively conserved in plants. The SaSAMS gene was cloned from a mangrove tree S. alba for the first time and this would facilitate further investigation on molecular mechanism of stress tolerance and regulation networks of stress-related genes in S. alba.

Squalene synthase gene cloning and analysis of Camellia oleifera based on RNA-Seq

2016/1/20 18:33:54
Camellia oleifera is a very important woody oil tree,the content of squalene is one of most important factor to evaluate quality of in the C. oleifera oil,to increase the content of squalene from molecular biology method. Changes of SQS related Unigene among seeds development were analyzed based on RNA-Seq,changes of the suqalene content in C. oleifera oil among seed development also tested. The result showed the same variation law. So SQS gene RACE primers were designed according to Unigene,and the full length SQS of C. oleifera was cloned,including 1 490 bp nucleotide,and the ORF is 1 266 bp long,encoding 422 amino acids sequence,which was blasted with that of other plants,the phylogenetic tree were mapped and analyzed. Then bio-informatics of SQS were studied,including structure,physicochemical properties,amino acid composition,and trans-membrane domain,signal peptide recognition,phosphorylation sites,function of secondary structure and domain analysis. The gene expression changes from August to October were similar to those of squalene content in seeds. It is concluded that the gene sequence encodes to be SQS of C. oleifera theoretically, which was a basis for the further study of functional verification and improving suqalene content in C. oleifera seed oil.

Effects of medium components and subculture period on plantlet regeneration of creeping bentgrass

DENG Yi-Fei, ZHANG Lu-Na, WU Yi-Cheng, CHEN Qin, DENG Zhi-Rui — 2016/1/20 18:33:54
Mature seeds of creeping bentgrass were used as explants to study the effects of 2,4-D concentration,combination of 2,4-D and 6-BA,and sucrose concentration on callus induction and the effects of subculture time,6-BA concentration,sucrose concentration on plantlet regeneration. The results showed that combination of 2 mg·L^-1 2,4-D and 0.1 mg·L^-16-BA produced the best result with 94% callus induction rate,and 30 g·L^-1 sucrose gave the best result,callus induction rate being 82%. During regeneration process,1 mg·L^-16-BA could bring the best differentiation frequency(62%)and 40 g·L^-1 sucrose resulted in the highest plantlet regeneration frequency(52%).The callus after the second time subculture(40 d after explants were laid on medium)gave the highest differentiation frequency(71%)which began to decline as subculture prolonged. After the fifth time subculture(100 d),only 18% differentiation frequency was obtained.

Establishment of an efficient regeneration system of Jijing88

2016/1/20 18:33:54
Jijing 88 was used as the research material and different media and hormone combinations were selected,in order to obtain its optimal in-vitro regeneration system. The results showed that NMB medium was the most suitable medium for rice callus induction among the 5 medium. The most suitable plant hormone ratio was investigated by using orthogonal test L₉(3³),and the induction rate was the highest when 2.0 mg/L 2,4-D,0.2 mg/L 6-BA,0.10 mg/L NAA were added. The differentiation rate and regeneration rate were the highest when 2.0 mg/L6-BA,0.5 mg/L NAA were added.

Clone and stress expression analysis of iron superoxide dismutase from Isatis indigotica

2016/1/15 2:06:44
Superoxide dismutase is a scavenger of superoxide anion free radical in vivo. They can effectively prevent reactive oxygen species damage to organism. An iron superoxide dismutase gene was isolated from Isatis indigotica, and was named as IiFeSOD. The full-length cDNA of IiFeSOD contained a 834 bp open reading frame(ORF)and encoded 277 amino acids, which might locate in mitchondria. Real-time PCR analysis showed that IiFeSOD had the highest expression in leaves,then in stems,and the least in roots. The expression level of IiFeSOD could be induced by NaCl. Levels of IiFeSOD transcripts could increase at three days after salt stress. However,the expression of IiFeSOD first increased and then decreased after salt stress at 100 mmol/L. Activity of SOD was also induced by NaCl and showed the analogous change rule with gene expression. Studies showed that massive accumulation of SOD was closely related with the stress tolerance of plants.

Effects of exogenous spermidine on the expression of HSP70 in cucumber roots under waterlogging stress

ZHANG Jian, LIU Mei-Yan, SHEN Jie, LIU Dan, YU Li-Xuan, LI Xue-Hua — 2016/1/15 2:06:44
RT-PCR and Western Blot were adopted to study the effects of exogenous spermidine(0. 5 mmol/L)on HSP70 in mRNA and protein expression of cucumber roots under waterlogging stress. The results showed that under waterlogging stress,the expression of HSP70 in cucumber roots soaked with distilled water at mRNA and protein level increased and then decreased with the extension of waterlogging time,when the waterlogging time was up to 4 h,the expression quantity of HSP70 mRNA and protein were extremely higher compared with no waterlogging treatment; the expression of HSP70 in cucumber roots soaked with exogenous spermidine at mRNA and protein level increased within 24 h,when the waterlogging time was up to 24 h,the expression quantity of HSP70 mRNA and protein were extremely significant different compared with no waterlogging treatment. Under waterlogging stress,the expression of HSP70 mRNA and protein cucumber roots soaked with exogenous spermidine were extremely higher when the waterlogging time was up to 12 h and 24 h compared with soaked with distilled water. Exogenous spermidine could induce the expression of HSP70 and protein in cucumber roots increased under waterlogging stress,and could relieve the harm caused by waterlogging stress.

Influence of enhanced UV-B radiation on F-actin of wheat mesophyll nucleus

2016/1/15 2:06:44
Nuclei which were treated after enhanced UV-B radiation were taken as materials in this research. The nucleus actin of wheat mesophyll was researched by fluorescent with FITC-ph and flow cytometry. The results showed that F-actin truly existed in the wheat mesophyll nucleus,which increased in its content after treated with the enhanced UV-B radiation. The FITC-ph fluorescent labeling displayed that the distribution and the content of the nucleus F-actin. We found that F-actin in the nucleolar was higher than the other areas in the wheat mesophyll nucleus. We speculated these were closely with “partition-bundle division”.

Changes of PS Ⅱ chlorophyll fluorescence characteristics in leaves of Arabidopsis thaliana plants with alternative oxidase under different light conditions

2016/1/15 2:06:44
Several chlorophyll fluorescence parameters were investigated under different light conditions(133,165,204,248,297,580,922,1 175,1 465 and 1 855)by a chlorophyll fluorometer of alternative oxidase gene overexpression lines(XX-2)and wild-type(WT)lines of Arabidopsis thaliana plants. The results showed that the parameter of maximal PSII quantum yield(Fv/Fm)didn't have significant change in two type leaves. The parameters of effective PSII quantum yield(Y(II))and coefficient of photochemical quenching(qp)decreased according to the increase of light intensity; the Y(II)and qp of XX-2 leaves increased at the same light intensity compared with WT. The parameters of coefficient of nonphotochemical quenching(NPQ)and quantum yield of regulated energy dissipation[Y(NPQ)]increased with elevation of light intensity; the NPQ and Y(NPQ)of XX-2 leaves declined at the same light intensity compared with WT. The quantum yield of non regulated energy dissipation[Y(NO)] declined with the aggrandizement of light intensity; the Y(NO)of XX-2 leaves decreased at the same light intensity compared with WT in the range of 165-1 855 μmol·m^-2·s^-1. The parameter of electron transport rate increased at the early stage of light treatment,the highest was found at 580 μmol·m^-2·s^-1,then declined according to the increase of light intensity; the ETR of XX-2 leaves increased at the same light intensity compared with WT.

Synthetical evaluation based on effects of pollination on fruit quality of ‘Red Fuji' apple

XU Chen-Shan — 2016/1/15 2:06:44
‘Red Fuji' Nagafu No. 2 was used as female parent and nine varieties were used as male parents to study the effects of pollination treatments on mature fruit of ‘Red Fuji' apple by synthetical evaluation function based on principal component analysis. The results showed that there were significant differences on main economic characters of fruit such as anthocyanin,fruit weight,vitamin C,ratio of TSS to acid,soluble sugars,soluble protein and titratable acidity. However,there was no obvious difference on fruit shape index and firmness. Six principal components(PC)whose total cumulative contribution reached 96.02% were extracted by principal component analysis. The synthetical evaluation function was composed of scores of principal components and relative variance contribution. From high to low,the arranging order of the synthetical scores of the nine pollination treatments,which were assessed by the synthetical evaluation function,was ‘Xueqiu',‘Meihong',‘Huoyan',‘Starking',‘Daoge',‘Quanjiahong',‘Xuanli',‘Hehong' and ‘Fenya'.

Chemical constituents from the leaves of Garcinia oblongifolia

2016/1/15 2:06:44
The compounds of the leaves of Garcinia oblongifolia were isolated by column chromatography,and their structures were inentified by the basis of physicochemical evidence and spectroscopic analysis. Eleven compounds were obtained and identified,which were Iisophytol(Ⅰ),α-Tocospiro B(Ⅱ),hendecane(Ⅲ),Palmitic acid(Ⅳ),(+)-(3R)-3-hydroxyl-4,4-dimethyl-4-butyrolactone(Ⅴ),Benzoic acid(Ⅵ),3,4-Dihydroxy-acetophenone(Ⅶ),Trans-p-hydroxycinamic acid(Ⅷ),4-Hydroxy 3-methoxy-benzoic acid(Ⅸ),β-Sitosterol(Ⅹ),Daucosterol(Ⅺ). All the compounds were isolated from this plant for the first time.

Activity of α-amylase inhibitor of chickpea seeds during their germination and maturation

2016/1/15 2:06:44
Inhibitory activity of α-amylase from chickpea seeds(embryo and cotyledon)during germination and maturation against α-amylase derived from 5 different sources were determined. The results showed that α-amylase inhibitor mainly exists in the embryo. During germination,1 d after germination,inhibitory activity of embryo extracts reduced to 9.9% against AOA,and there was no inhibitory activity to HSA,PPA and PA; inhibitory activity of cotyledon extracts against AOA and HSA reduced to 21.5%,28.3%,respectively,and there was no inhibitory activity to BSA and PPA. 4 d after germination,both cotyledon and germ extracts on 5 sources were undetectable inhibitory activity. During seed maturation,both cotyledon and embryo extracts on the activity of α-amylase inhibitor increased with the days of seed maturity and reached to the maximum on 30 d,and the inhibitory activity of embryo extract was higher on HSA and PA,48.9% and 47.5%,respectively.

Isoflavone content change research of dwarf mutant soybean

2016/1/15 2:06:44
The isoflavones of stems,leaves and seeds were detected by flow injection chemiluminescene method from the Dongnong 42 and two dwarf mutants(HK8 and HK11); at the same time,the flow injection chemiluminescence and HPLC were compared in this paper. The results of the study showed that the isoflavone content could be more simply and fleetly detected by flow injection chemiluminescene,the RSD was 0.69%-2.30%,and the recovery was 95.5%-104.5%. It can be concluded that the isoflavone can be accurately detected by flow injection chemiluminescene method in this experiment; the isoflavone is proportional to the plant height; the isoflavone content of seeds is higher than that of stems or leaves.

Effects of Se on photosynthetic characteristics of Siraitia grosvenorii seedling under Cd²⁺ stress

2016/1/15 2:05:15
The tissues cultured seedlings of Siraitia grosvenorii were used as the material which were planted in the indoor cultivation and the soil with nutrition bought from market. The cultured seedlings of S. grosvenorii were added different concentrations of Cd²⁺(0,10,50,100,200 mg·kg^-1 )and Se²⁺(1 mg·kg^-1)into mediums under sterile conditions. The correlative photosynthetic characteristics indices were analyzed after cultivating 20 d. The results showed that the low concentration Cd²⁺had the effect on the contents of chlorophyll,photosynthetic rate(Pn),transpiration rate(Tr)and stomatal conductance(Gs)were not big or increased slightly. But under high concentrations treatment of Cd²⁺,the contents of chlorophyll,photosynthetic Rate(Pn),transpiration rate(Tr)and stomatal conductance(Gs)of leaves decreased obviously. With the increase of Cd²⁺ concentration,the rising trend appeared in leaf intercellular CO₂ concentration,the addition of Se delayed the decline of chlorophyll,and made photosynthetic rate(Pn),transpiration rate(Tr),stomatal conductance(Gs)increased,but reduced the leaf intercellular CO₂ concentration(Ci). It was shown that the toxicity of high concentrations of cadmium ions could also hurt photosynthetic characteristics of S. grosvenorii seedling,which,therefore,affected the growth of S. grosvenorii seedling. The mixed treatment of cadmium and selenium reflected that selenium had alleviated function on the toxicity of cadmium.

Effects of two exogenous chemical substances on the osmotic stress-induced damages to rice roots

2016/1/15 2:05:15
The effects of the scavenger of H₂O₂ and specific inhibitor of alternative oxidase on the osmotic stress-induced damages to rice roots were studied. The results showed that PEG 6000 inhibited the growth of rice roots,decreased the relative water content,inceased the H₂O₂ content,and caused cell death. Pretreatment with 5 mmol·L^-1 dimethylthiourea(DMTU),a scavenger of H₂O₂,significantly decreased the increment in the H₂O₂ content and partially alleviated the root cell death and the decrease of the relative water content induced by PEG 6000. However,DMTU had little effect on the growth of rice roots exposed to PEG stress. Under the PEG-induced stress,pretreatment with 1 mmol·L^-1 salicylhydroxamic acid(SHAM),a specific inhibitor of alternative oxidase,significantly decreased the growth of rice roots and relative water content,but increased the levels of H₂O₂ content and cell death. The results suggested that DMTU could alleviate the osmotic stress-induced damages to rice roots,whereas SHAM could intensify those damages.

Comparative research of reactive oxygen species metablisms of the CMS line and the maintainer lines in Kenaf(Hibiscus cannabinus)anther

2016/1/15 2:05:15
Kenaf(Hibiscus cannabinus)anther of the cytoplasmic male-sterile(CMS)line L23A and its maintainer line L23B were employeded as experimental materials to reveal the relationship between CMS and metabolisms of reactive oxygen species(ROS)by observing changes of starch and soluble sugar contents and comparing productions of superoxide anion radical(O^-₂·), respiratory rate as well as content of malonadehyde(MDA)and active oxygen exterminating enzymes(SOD and POD). The results showed that there was no evident difference between respiratory rate of CMS line and that of maintainer line at mononuclear stage, but anther of CMS line had higher content of O^-₂· than that of maintainer line at this stage. Respiratory rate in anther of the CMS was lower than that of maintainer line at binucleate pollen stage, while O^-₂· content in sterile anther approximated to that in fertile anther of maintainer line. There was no evident change in respiratory rate of CMS line at mononuclear stage and binucleate pollen stage, but respiratory rate in anther of maintainer line obviously increased from mononuclear stage to binucleate pollen stage. With the growth of anther, big starch grains content in L23A connective did not change during its microsporogenesis and soluble sugar content in L23A was lower than that of L23B at both of mononuclear stage and binucleate pollen stage. This possibility gave rise to the speculations that the decline of cyanide-resistant respiration in anther lead to disturbance of material metabolism and energy metabolism and starch grains presented in the connective tissue were not utilized by anther of the CMS line, meanwhile the intracellular surplus electrons were not effectively passed on to O₂ by cyanide-resistant respiration, which leads to increase O^-₂· content in anther of CMS line so that there is higher MDA content in anther of CMS line at mononuclear stage and binucleate pollen stage. Simultaneously, POD activity was lower than that of its maintainer line at mononuclear stage and binucleate pollen stage. While activitiy of SOD was higher than that of its maintainer line at mononuclear stage, but lower than that of its maintainer line at binucleate pollen stage. During the abortion of CMS anther, O^-₂·and MDA excessively accumulate in CMS anther and activity of SOD and POD would decline,which disrupted the balance between the production and elimination of ROS and made pollen mother cells apoptosis.

Cloning and sequential analysis of phytoene synthase (PcPSY1)gene in Pogostmen cablin

2016/1/15 2:05:15
Phytoene synthase(PSY)is a key enzyme in plant terpenoid biosynthetic pathway. According the transcriptome dataset of Pogostmen cablin, one unique sequence encoding PSY was discovered. The primers were designed according to the transcript sequence of PcPSY1 from the P. cablin transcriptome dataset with Primer 3 online. The open reading frame of PcPSY1 was cloned using RT-PCR strategy. The cDNA named as PcPSY1 contains a 1 550 bp open reading frame and encodes a predicted protein of 439 amino acids. The GenBank accession number for this gene is KC862310. PcPSY1 has no signal peptide. PcPSY1 is highly homologous to other PSY protein in 21 different species and is more related to that from Osmanthus fragrans. This study cloned and analyzed mechanism of terpenoid biosynthesis in Pogostemon cablin plants.

Polymorphic microsatellite markers from expressed sequences tag in Nicotiana tabacum for obtaining the genes of preponderant expression in zygotes

2016/1/15 2:05:15
In order to obtain the genes of preponderant expression in zygote cells of tobacco,CAP3 program was used to assemble ESTs from GenBank,and MISA program were used to screen microsatellites. Then polymorphic microsatellites were amplified in cDNA of tobacco zygotes library. The results showed that sixteen polymorphic SSR markers from tobacco ESTs deposited in a public sequence database were developed. Of these,nine genes could be successfully amplified in cDNA of tobacco zygotes library. The study would provide new channel to screen genes of preponderant expression in tobacco zygotes.

Multiplex PCR analysis of Indica-japonica differentiation of the chloroplast DNA in weedy rice

2016/1/15 2:05:15
The whole chloroplast genomes of the Indica cultivar 93-11 and Japonica cultivar Peiai64S were analyzed and compared. A multiplex PCR marker of the chloroplast DNA ORF100 and ORF29-TrnC^GCA region was optimized and constructed. More than 200 weedy rice,Asian cultivated rice and common wild rice accessions were then tested using the multiplex PCR. The results showed that there were obvious chloroplast DNA indica-japonica differentiations amongst the weedy,Asian cultivated and common wild rice accessions. Furthermore,the differentiations in weedy rice were correlated to the collected regions that were in accordance with the south indica type and north japonica type of cultivated rice in China. It was suggested that the japonica line of weedy rice should evolve from degraded cultivated rice or the japonica varieties(as the female parent)natural hybridization with other oryza materials for there was no wild form in north China.

Developing SSR primers in Bombax ceiba(Bombacaceae)

2016/1/15 2:05:15
Bombax ceiba is a multipurpose tree species of tropical forests. Nine novel polymorphic microsatellite loci for B. ceiba were isolated and characterized through the combined biotin capture method. Markers were tested on 32 individuals collected from dry-hot valleys in southwest China. The number of alleles ranged from 2 to 6. The expected and observed heterozygosity values ranged from 0 to 0.525 and 0.121 to 0.561,respectively. One locus showed significant deviation from Hardy-Weinberg Equilibrium. No significant linkage disequilibrium was found. The microsatellite markers that were characterized in this study were the first to be developed for B. ceiba and would have great potential for different genetic studies of wild populations of this species.

Preliminary study on relationship of common buckwheat quality with ecological factors and agronomic traits

2016/1/15 2:05:15
Variations of buckwheat grain quality(flavonoid and protein contents)and the correlations of the quality and yield with ecological factors and agronomic traits were studied using 8 common buckwheat varieties in 19 different locations across the country. The results showed that there were significant variations of seed flovonoid content and protein content among different varieties and different locations of the same varieties. There were all significant relationships of yield with plant height,grain numbers of per plant,and grain weight of per plant,of the flavonoid content with the elevation and the average temperature of the growth period,of the protein content with latitude,and their correlation coefficient were 0.271^*,0.588^**,0.495^**,0.270^*,-0.330^** and 0.212^*. The path analysis indicated that grain number per plant the key trait for yield,and the higher the grain number per plant, the higher the yield. The average temperature in the growth period was the key factor of the flovonoid content,and the suitable lower average temperature in buckwheat fertility period could increase the accumulation of seed flavonoid. Latitude was the key trait affecting the protein content,and higher latitude would help to increase the protein content. This study also showed that different varieties had different sensitivities to ecological factors.

Seasonal variation in photosynthesis of an introduced bamboo species Guadua amplexifolia

2016/1/15 2:05:15
Photosynthetic characteristics and main impact factors were measured in different seasons,and the data were analyzed by correlation and path coefficients,and different irradiances were measured in Hua'an County,Fujian Province,in order to study Guadua amplexifolia's photosynthesis characteristic,CO2 assimilation rate on different times in one day. The results indicated that light saturation point(LSP),light compensation point(LCP),dark respiration rates(Rd),and net photosynthesis at saturation light(Pn_max)all reached highest in July,lower in April and October,the lowest in January. In all seasons but April,daily course of net photosynthesis rate(Pn)had midday-depression pattern. The daily mean value of Pn was in the sequence April>July>October>January. All environmental and physiological factors except for relative humidity(RH)had significant correlation with Pn,such as Tr,Ci,Gs,VPD,PAR,Tleaf and so on. The main environmental factors differed in every season. In January,leaf temperature(Tleaf)was the most important factor to Pn. But the effect of RH increased obviously in October. The effect of tested environmental factor to Pn was in the sequence VPD>PAR>RH>Tleaf in the whole growth period.

Allelopathtic effects of Phragmites communis aqueous extract on seed germination and seedling growth of Phalaris arundinacea

FU Wei-Guo, TIAN Yuan-Fei, TANG Juan-Juan, YIN Qi-Lin — 2016/1/21 12:11:49
Allelopathtic effects of Phragmites communis on Phalaris arundinacea was explored by studying the effects of aqueous extract of Phragmites communis with different concentrations(CK: 0; Treat 1:150 g/L; Treat 2:300 g/L)on seed germination and seedling growth of Phalaris arundinacea. The results showed that with the increase in the concentration of aqueous extract from Phragmites communis,seed germination rate,seed germination index,seedling height,seedling taproot length,and chlorophyll content in leaf showed significant or extremely significant reduction,while allelopathic effect indices based on the above indicators showed significant or extremely significant ascension. Activity of antioxidant enzymes of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT),and content of malondialdehyde(MDA)showed varying degrees of upward or downward trend with the increase in the concentration of aqueous extract. All these results indicated that allelopathic stress effect of Phragmites communis on Phalaris arundinacea was becoming more and more obvious with the increase in the concentration of extracts of Phragmites communis.

Physiological effects of manganese on the germination of sweet corn

CHENG Xin-Xin, LIU Zheng — 2016/1/21 12:11:49
Physiological effects of manganese on the germination of sweet corn were explored in this paper. The results showed that the differences of physiological indicators were significant on two genotypes of sweet corn after Mn²⁺ soaking. The rate of seed germination of sweet corn did not change significantly. Germination potential and vigor index of Lvsechaoren increased significantly. The effect of treatment with the concentration of 1.00 g/L was the best. The dethydrogenase activity of Lvsechaoren and Huatian 01 increased with increasing concentration. The POD activity and CAT activity were on the rise and the MDA content of seed germination decreased under the proper concentration of Mn²⁺. In addition,the soluble sugar contents which were treated after Mn²⁺ soaking were not significant than that of untreated,but the difference between two varities was large. So the role of Mn²⁺ soaking was significant in different genotype of sweet corn.

Effects of low-energy N⁺ ion beam irradiation on the main physiological characteristics of rice(O.sativa) with different chromosome sets in seedling period

2016/1/21 12:11:49
Three different doses of low-energy N⁺ ion beam were injected into the embryo of diploid rice seeds and corresponding autotetraploids. The germination capacity,seedling rate and five major physiological indexes superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),malonaldehyde(MDA),and soluble sugar were examined preliminarily at seedling period. The results showed that with the increase of the implantation dose,the germination capacity and seedling rate of experimental materials tended to decline,and the difference between each treatment group and CK reached the particularly significant level(P<0.01)under the maximum dose(5.0×10¹⁷N⁺/cm²). The SOD,POD and CAT activity of diploid materials increased first and then decreased with the dose increasing,and all of them reached the maximum at the dose of 3.0×10¹⁷N⁺/cm²,while the SOD,PODand CAT activity of tetraploid materials showed increase-decrease-increase curve and all of them reached maximum at 1.0×10¹⁷N⁺/cm². The MDA content of diploid materials decreased first and then increased with the dose increasing,while that of tetraploid materials showed decrease-increase-decrease curve. There was no particularly significant difference among the treatment group of soluble sugar content,but the content of soluble sugar was lower than that of the CK. Thus,different doses of low-energy N⁺ ion beam had a significant effects on the germination capacity,seedling rate and main physiological characteristics of rice with different chromosome sets in seedling period,and there were also some differences between materials with different ploidies.

Correlation analysis on nutrition status of soil and leaf nutrients for Pyrus pyrifolia cv.Hosui citrus demonstration orchard

2016/1/21 12:11:49
Bearing trees of Pyrus pyrifolia cv.Hosui in the northern area of Guilin,where physiologically abnormal phenomena were very typical,were selected as the research objects to study the relationship between nutrition of leaves and soil nutrients and its dynamic change rule in different periods by mensurating mineral nutrient contents of leaves and soil in the annual growth cycle,so as to probe into the relationship between the nourishments of leaves and soil in annual growing season. The results were as follows:(1)N,P and K contents of the leaves were abundant in the whole growing season,and the variation range of P contents was the most,while that of N,K contents less,and did not reach a significant level(P>0.05).(2)The soil organic matter,hydrolysis of N contents of the pear gardens were very abundant,and effective P and available K contents were generally high. Available N,P,K contents of the soil in the growth cycle reached a significant level(P<0.05),and the soil organic matter content was relatively stable.(3)N contents in the leaves had a significant positive correlation with soil organic matter,hydrolysis of N,effective P,and available K content of the soil(P<0.05),and P and K contents in the leaves significant negatively and positively correlated with soil hydrolysis of N respectively(P<0.05),while had insignificant relationship with soil organic matter,effective P and available K contents of the soil.

Overexpression of salicylate hydroxylase NahG declines resistance to Ralstonia solanacearum infection in tobacco

2016/1/21 12:11:49
Studies showed that the plant expression vector of pMDC32-NahG was construed in tobacco K326,and obtained NahG-OE T1 lines. Comparing to wild type K326,NahG-OE lines did not exhibited any phenotypic difference,but exhibited higher susceptibility to Ralstonia solanacearum inoculation. Meanwhile,it was found to be coupled with up-regulated expression of JA-depended NtPR1-b,and down-regulated expression of SA-depended NtPR3 and NtPRQ. Results suggested that SA-degrading NahG tobacco plants declined resistance to R.solanacearum infection

Studies on tissue culture and rapid propagation technique of Echinodorus subalatus

2016/1/21 12:11:49
The in vitro culture of Echinodorus subalatus was established by using the unelongated stem as explants. The method of surface sterilization of explants,the effects of different plant growth regulators on the multiplication and rooting were investigated. The results showed that 20% sterile explants could be obtained by twice surface treatment with 0.1% HgCl₂ for 8 min and 6-8 min with an interval of 24 h. When cultured on a 1/2 MS multiplication medium supplemented with 6.0 mg·L^-1 of 6-BA for 42 d,the test-tube seedling exhibited a multiplication coefficient of 9.3. For optimal rooting,induction on a 1/2 MS rooting medium supplemented with 0.2 mg·L^-1 of NAA for 28 d could produce rooted plantlets with 100% rooting rate,and each plantlet generated about 18 roots on average. Rooted plantlets were acclimatized and transferred into outdoor aquatic environment with 100% survival rate.

Establishment and optimization of ISSR-PCR reaction system on Zanthoxylum dissitum

2016/1/21 12:11:50
In order to establish the optimal ISSR-PCR reaction system of Zanthoxylum dissitum,several factors that mostly affect the amplification of ISSR-PCR reaction of Z.dissitum,including DNA template,Mg²⁺concentration,primers,dNTPs,TaqDNA polymerase dosage and annealing temperature,were investigated and optimized by single-fact test and orthogonal-design method in the experiment. The results indicated the most stable and suitable reaction system had been established when the amount of DNA template was 60 ng,the dosage of Taq DNA polymerase was 2.4 U and the concentration of MgCl₂,dNTPs and primers was 2.5 mmol/L,0.15 mmol/L,0.6 μmol/L respectively. Further more,18 ISSR primers that possessed the characteristics of stable amplification and rich polymorphism were screened from 100 ISSR primers,and they were examined by the germplasm test of 10 Z.dissitum parts. The results of the examination proved that this ISSR reaction system had the advantages of clearness,stability and good repeatability. In conclusion,this established ISSR-PCR reaction system provided an available and reliable method for genetic diversity analysis and germplasm classification of Z.dissitum.

Construction of OsRUS 2.1 yeast twohybrid prey vectors and detection of their toxicity and selfactivated activity

2015/12/15 14:36:46
Four fragments of rice(Oryza sativa)ROOT UVB SENSITIVE 2.1(OsRUS2.1)，OsRUS2.1(11317)，OsRUS2.1(1138)，OsRUS2.1(139879)，OsRUS2.1(8801317)，were amplified by PCR from cloned OsRUS2.1 plasmid，and were ligated with pMD18Tsimple vector，then transformed to E.coli TOP10 competent cell. The positive clones were selected and sequenced. The confirmed fragments were subcloned to prey vector pGADT7. The four constructed pGADT7 prey vectors were further confirmed by enzyme digestion and sequencing. The confirmed 4 types of pGADT7 prey vectors were transformed to Y187 yeast competent cells. The selfactivation and toxicity of the plasmids to host yeast Y187 were detected by LacZ and MEL1 activity assays and culturing in auxotroph medium SDLeu DO． Results showed that the four constructed plasmids had no selftranscriptional activity and not toxicity to yeast strain Y187，and they could be used in the following yeast twohybrid experiments．

Bioinformatics analysis of CBF2 in three different chilling resistance grapes and construction of plant expression vector

ZHANG ZheMin， SUN Ping， WANG WangTian， LI MengFei， LI Wei*， WANG YaLin — 2015/12/15 14:36:46
The fulllength gene of Crepeat binding factor 2 was amplified from the genomic DNA in three different chilling resistance grapes:Cabernet sauvignon，Beta and Vitis amurensis. The methods of bioinformatics were also applied to analyze the sequencing results. Sequencing results showed that the cloned CBF2 gene in three different chilling resistance grapes had 969 bp and encoded 253 amino acids，and had 96.7%-97.94% identity with the sequence published on GeneBank. The bioinformatics analysis showed thatCBF2 from three different chilling resistance grapes had continuous opening reading frame，and the deduced amino acid sequence also had AP2 binding domain，so we should have preliminary judegement of cold resistance effect of cloned CBF2. But the analysis from the homology of amino acid sequences，phylogenetic tree，physicochemical property and hydrophobicity and hydrophilia showed that there were some differences among the high chilling resistance grape Vitis amurensis and Beta，and the low chilling resistance grape Cabernet sauvignon.The comparison of sequence homology with the sequence published on GeneBank showed that the worst one was C.sauvignon；and the 7 amino acids from N terminal in C.sauvignon were totally different with Vitis amurensis and Beta；the difference might be related to the higher hydrophobicity in the head of amino acid residues from Cabernet sauvignon. Based on the vector pBI121 with promoter 35S，the plant expression vector pBI121CaMV35SCBF2 was constructed successfully，which could offer the basic researching data to improve the coldtolerance ability of grape.

RACE cloning and bioinformatics analysis of nitrate reductase in Beta vulgaris

2015/12/15 14:36:46
Nitrate reductase is an important and limited enzyme in nitrogen metabolism. The functional research of nitrate reductase plays a key role in increasing sugar beet(Sb) yields. In this study，SbNR gene was cloned by RACE and its 3D structure and physiological function were analysed by bioinformatics and expressed in Arabidopsis thaliana to show the gravity. The results were as follows:the full length of SbNR gene was 2760bp；SbNR was soluble and hydrophobic；secondary structure analyses showed that SbNR belonged to mixed protein；SbNR was a new member of NR family，containing molybdopterinbinding，cytochromebinding，FADbinding and NADbinding domain. It had no signal peptide，but one transmembrane domain in C terminal，and the mutants showed later response to the gravity，and which confirmed that NO involved in plant’s gravity.

Response of yield，nitrogen uptake in Jiyou716 to nitrogen management under different tillage patterns

2015/12/15 14:36:46
Characteristics of rice yield formation and nitrogen uptake under different soil tillage patterns and nitrogen fertilizer management patterns were studied, which provided theoretical basis for rice rational fertilizer application under different tillage systems. With Jiyou716 as materials for field experiments，change of yield，nitrogen uptake in rice at three N rates(N0，N1，N2) and two fertilizering patterns(F1，F2) under notillage and conventional tillage were studied. The results indicated that response of yield and nitrogen uptake of rice to nitrogen amount were basically similar under notillage and conventional tillage，but inconsistent to nitrogen fertilizering pattern. With N rate increasing, the yield，dry matter accumulation and N accumulation of rice increased，and N production efficiency，dry matter harvest index and N agronomy efficiency decreased. Heavy earring fertilizer was better for N accumulation and dry matter accumulation increase under notillage and conventional tillage. But inconsistent response for grain yield. Heavy earring fertilizer was in favor of yield increase under notillage while heavy basic fertilizer and tillering fertilizer was better for yield increase under conventional tillage.

Effects of ALA on physiological characteristics of Cassia obtusifolia seedlings under salt stress

2015/12/15 14:36:46
In order to get the method to improve the salt resistance of seedlings for Cassia obtusifolia under salt stress，physiological characteristics of C.obtusifolia seedlings were studied. Several physiological indexes of C.obtusifolia seedlings under salt stress like photosynthetic pigment，superoxide anion（O〖ST4.〗2〖HT〗.），malondialdehyde(MDA)，superoxide(SOD)，peroxidase(POD) and catalase(CAT) were measured. The rate of decrease was relieved and the content of O〖ST4.〗2〖HT〗.，MDA decreased. The activities of three enzymes including SOD，POD and CAT were increased by the treatment of ALA，and reached the maximin(80.0 U•g1，196.4 U•g1 and 114.1 U•g1 respectively)with the concentration of 50 mg•L1. ALA with concentration of 50 mg•L1 could significantly alleviate the damages to the seedlings of C.obtusifolia under salt stress，and promote the salt resistance of the seedlings.

不同甘蔗品种叶片气孔对水分胁迫的响应

2015/12/15 11:54:15

盐胁迫对盆栽滁菊生理特性的影响

2015/12/15 11:54:15

赤桉单萜合成酶基因的分子克隆与生物信息学分析

2015/12/15 11:54:15

柚木无性系愈伤组织诱导及植株再生

2015/12/15 11:54:15

Cloning and expression analysis of mads3 in maize CMS-C sterile line and its maintainer line

2015/12/15 11:20:20
MADS-box genes involve in various developmental processes in plant,and play an important role in the regulation of floral development. In this paper,mads3,a member of MADS box gene family was cloned by RT-PCR in maize. Sequence analysis showed that 8 sits single-base substitution mutations in CDS region were found in CMS-C line in comparison to maintainer line,which may result in 5 individual amino acides change. Furthermore,real-time qPCR analysis showed that mads3 was significantly down-regulated in CMS-C line at tetrad stage,and was significantly up-regulated at uninucleate stage. These results laid the foundation for further studying the relationship between MADS-box genes and CMS-C in maize.

Effect of different nitrogen application modes on root growth,rhizosphere soil characteristics and rice yield under no-tillage

2015/12/15 11:20:20
The effect of nitrogen application modes on root growth,rhizosphere soil characteristics and rice yield under no-tillage and cast transplanting rice was studied. The results showed that balanced fertilization made obvious influence on no-tillage and cast transplanting rice root growth,and physiology. The rice shoot dry weight and biomass per plant,root number,root density,root activity of balanced fertilization were higher than other treatments,the differences were significant in rice later growth. However,there were no significant differences in root length. Balanced fertilization increased spike lets per panicle and seed set. Balanced fertilization significantly increased 0-10 cm soil layers organic,available nitrogen and urease activity of 0-2 mm rhizosphere soil in rice late growth. Therefore,it can been concluded that balanced fertilization acquired higher grain yield by improving rhizosphere soil,promoting root growth and increasing effective panicles.

Effects of low temperature stress on antioxidant enzymes activities in the subcellular of two Sabina species

2015/12/15 11:20:20
Seedlings of Sabina przewalskii and S. chinensis were used to study the effects of low temperature stress on antioxidant enzymes activities and subcellular distribution in the leaves of Sabina. The results showed that: under low temperature stress, the contents of malonaldehyde(MDA)and antioxidant enzymes activities increased firstly and then decreased, and compared with Sabina chinensis, the more species and higher activities of antioxidant enzymes were found in S. przewalskii, but MDA contents of S. przewalskii were lower than S. chinensis,it suggested that S. przewalskii showed a prevailing strategy than S. chinensis in order to develop the tolerance to freezing. Moreover,the localization of superoxide dismutase(SOD)and ascorbate peroxidase(APX)in the leaves of two Sabina species was chloroplast>cytosol>mitochondria,the localization of catalase(CAT)was mitochondria>chloroplast>cytosol,the localization of glutathione reductase(GR)was mitochondria>cytosol>chloroplast,the localization of peroxidase(POD)in S. przewalskii was cytosol>chloroplast>mitochondria,the localization of POD in S. chinensis was cytosol>mitochondria>chloroplast,moreover subcellular distribution of antioxidant enzymes(SOD,APX,GR)in organelles were different significantly,this showed that antioxidant enzymes played a major role in one of organelles,which provided theoretical basis for cloning of antioxidant enzymes gene in subcellular fractions.

Physiological response of water stress of Guizhou endemic plant Camellia delicate

2015/12/15 11:20:20
Camellia delicata is Guizhou endemic species and ornamental tree,so research on water stress is very important for the popularize. The paper studied 4-years seedlings,by principal components analysis method of physiological index,tested and evaluated the physiological response of waterlogged for 30 days and water stress. The results were as follows:(1)When the plants waterlogged,leaves grew well,there were no obvious changes in gloss and ornamental. The differences of leaf moisture content,MDA,SOD,POD and CAT were under 5%,soil moisture content increased 3.47%,cell membrane permeability decrease 1.59%. The damage to C. delicate short-term waterlogged was not obvious;(2)With drought time extension,new leaves were shriveled,old leaves broke off plant. Soil moisture content and leaf moisture content were negatively related to drought period,decreased slowly. The cell membrane permeability was positively correlated with drought period,grew slowly. But the plants returned to normal growth after watering 10d,and had not obvious physiological change response;(3)The SOD and MDA increased respectively 0.66% and 5.31%,POD fluctuated and increased 6.47% with drought time. CAT activity also increased 33.33% with drought 20 days,reached maximum 20.8 U·g^-1·min^-1. In the resistant physiology,which played the leading role in drought were leaf moisture content,POD,CAT.

Cold resistance mechanism study on effect of exogenous SOD under low temperature stress of young loquat fruits

2015/12/15 11:20:20
Young loquat fruits'activities of antioxidant enzymes,cell membrane permeability and content of malondialdehyde(MDA)after spraying different concentrations of Poly-aspartic Mangan(MSOD)under low temperature stress were studied. The results showed that MSOD with different concentrations(5.0,12.5,20.0 mg/L)could reduce cell membrane permeability and content of MDA and enhance the activities of superoxide dismutase(SOD),catalase(CAT),ascorbate peroxidase(APX)and peroxidase(POD),the concentration of 12.5 mg/L had the best effect.

Asymbiotic seed germination and high-frequency seedling regeneration of Spiranthes sinensis, an endangered orchid

2015/12/15 11:19:55
Seeds of Spiranthes sinensis were used to find out appropriate media for in vitro germination and seedling regeneration. Results showed that seeds could germinate on medium without plant growth substances,but failed to develop into seedlings; seedlings formed on medium with 1.0 mg·L^-1 KT,0.1 mg·L^-1 IAA and 0.1 mg·L^-1 GA₃; while seeds could not germinate on media containing lower concentration auxin(1.2 mg·L^-1 NAA). Seedlings could remain high proliferation rate(2.8 times)on media containing higher concentrations kinin and lower concentration auxin,they were eventually transplanted to greenhouse after cultured on hardening medium for 80 d. The suitable seedling proliferation and hardening media were 1/2MS+12.0 mg·L^-1 6-BA+0.1 mg·L^-1 NAA+10.0 mg·L^-1 adenine and 1/2MS+1.0 mg·L^-1 KT+0.1 mg·L^-1 IAA+10.0 mg·L^-1 adenine,respectively.

Study on rapid propagation and germplasm conservation in vitro of five species of genus Gynostemma in Guangxi

2015/12/15 11:19:55
Using stem with buds as explants,experiments were conducted to observe the effects of different concentration 6-BA and NAA 0.02 mg·L^-1on induction,differentiation,proliferation of Gynostemma in Guangxi,in order to establish their rapid propagation systems. Meanwhile,different concentrations of the mineral nutrition,sucrose and plant grow inhibitors(CCC,ABA,PP₃₃₃)were researched. The results showed that the best induction medium was MS medium supplemented with 2.0 mg·L^-16-BA and 0.02 mg·L^-1NAA. The suitable proliferation medium for G. compressum was MS+6-BA 2.0 mg·L^-1+NAA 0.02 mg·L^-1 and MS medium supplemented with 1.5 mg·L^-16-BA and 0.2 mg·L^-1NAA was best for others. The rooting medium was 1/2MS medium with 0.02 mg·L^-1 NAA,accounting for 100% of rooting rate. 1/2MS medium and sucrose 40 g·L^-1 were the most suitable for the conservation in vitro of all species. Plant grow inhibitors could slow down the grow speed effectively,and the best kind and concentration were ABA 1.0 mg·L^-1and CCC 1.0 mg·L^-1. Several species were inhibited by CCC 1.0 mg·L^-1and the survival rate was up to 94.5% after being conserved 360 d. PP₃₃₃ was not suitable for all species to conserve in vitro. Vigor testing showed that the ability of propagating and rooting of the plantlets didn't fall down.

Relationship between post-anthesis dry matter remobili- zation from culm, leaf in different position and grain growth of Indica rice varieties Baguixiang and Guihuazhan

2015/12/15 11:19:55
Field experiments were conducted to investigate carbon and nitrogen accumulation & translocation and grain growth after flowering with two indica rice Baguixiang and Guihuazhan as material in 2008 and 2009. The main results are as follows.(1)There were some differences in post-anthesis dry matter remobilization among leaves blade,sheath and internode of the quality rice in different positions. The contribution of the second leaf sheath from top(SS)to the grain was greater than the first leaf sheath from top(FS)and the third leaf sheath from top(TS)and the contribution of the third internode from top(TI)to the grain was more than the first internode from top(FI)and the second internode from top(SI);(2)Differences of the grain in various parts were presented in grain filling rate and initial grain filling potential. Filling rate and grain weight at the inflexion point claimed as: UPG(upper position grains)>MPG(middle position grains)>BPG(basal position grains). Upper position grain showed earlier initial filling potential,higher filling time rate while lower position grain lower filling rate,lagging filling time,and grain weight showed UPG>MPG>BPG;(3)Dry matter translocation rate and efficiency of leaf blade and sheath and internode post-anthesis were proportional to grain were correlated with initial grain filling potential. The corrational between organs and grain initial were displayed as follows:internode>leaf sheath>leaf,and the correlation coefficients in different parts showed inverted 2 from top>3 from top>1 from top. The effect of dry matter remobilization of internode on grain growth was much more than leaf sheath,and least of leaves. The first internode from top and lower position grain growth occurred with synchronicity. The same case happened to the second internode from top and the middle position grain,the third internode from top and basal position grain.

Allelopathic effects of Eucalyptus grandis&#215;E. urophlla leaf volatile on seed germination and seedling growth

2015/12/15 11:19:55
The allelopathic effects of Eucalyptus grandis&#215;E. urophlla plantation leaf volatile on surrounding crop were investigated,and seedling growth of three plants such as Zea mays,pepper,tomatoes were studied by using different quality fresh eucalyptus leaves and the eucalyptus essential oil from fresh leaves,the seed germination. The results showed that:(1)In three kinds of tested plants,E. grandis&#215;E. urophlla leaf volatiles had little effect on zea mays seed germination,great effect on pepper;(2)When the amount was less than 200 g,E. grandis&#215;E. urophlla leaf volatiles had no obvious effect on seed germination of three tested plants; When the dosage was 400 g,it could completely inhibit seed germination of peppers,tomatoes,and could reduce seed germination significantly(P<0.01);(3)With an increase of the dosage of leaf,the Zea mays growth first promoted and then suppressed,and it was significant(P< 0.05)or extremely significant level(P<0.01)to bud height,fresh weight,dry weight;(4)when the amount was less than or equoal to 100 g,the E. grandis×E. urophlla leaf volatiles was not obvious effect on seedling growth of peppers and tomatoes; when the amount reached 200 g,it could significantly inhibit the seedling growth peppers and tomatoes;(5)The eucalyptus essential oil had similar inhibitory effect on the 3 plants species with leaf natural volatile,and the effect was more significant.The allelopathic effects of Eucalyptus grandis&#215;E. urophlla plantation leaf volatile on surrounding crop were investigated,and seedling growth of three plants such as Zea mays,pepper,tomatoes were studied by using different quality fresh eucalyptus leaves and the eucalyptus essential oil from fresh leaves,the seed germination. The results showed that:(1)In three kinds of tested plants,E. grandis&#215;E. urophlla leaf volatiles had little effect on zea mays seed germination,great effect on pepper;(2)When the amount was less than 200 g,E. grandis&#215;E. urophlla leaf volatiles had no obvious effect on seed germination of three tested plants; When the dosage was 400 g,it could completely inhibit seed germination of peppers,tomatoes,and could reduce seed germination significantly(P<0.01);(3)With an increase of the dosage of leaf,the Zea mays growth first promoted and then suppressed,and it was significant(P< 0.05)or extremely significant level(P<0.01)to bud height,fresh weight,dry weight;(4)when the amount was less than or equoal to 100 g,the E. grandis×E. urophlla leaf volatiles was not obvious effect on seedling growth of peppers and tomatoes; when the amount reached 200 g,it could significantly inhibit the seedling growth peppers and tomatoes;(5)The eucalyptus essential oil had similar inhibitory effect on the 3 plants species with leaf natural volatile,and the effect was more significant.

Secretion of organic acids from root apices under Al stress in rye and wheat

2015/12/15 11:19:55
Secretion of organic acids from roots apices in rye and wheat were investigated by a system for in vivo root apex culture and micro-exudates collection,to elucidate the mechanisms for Al-induced secretion of organic acids. The results showed that Al(50,100,200,300 μmol·L^-1AlCl₃)induced secretion of citrate and malate from rye root apices,while wheat root apices secreted malate alone in response to Al treatment. Malate was significantly secreted from root apices of rye and wheat after the treatment with Al for 3 h,and malate was secreted with a high secretion rate within 9 h treatment. There was a lag between the secretion of citrate and Al treatment in rye. At initial 3 h,Al failed to induce significantly secretion of citrate. On the other hand,the addiction of anion channel inhibitor A-9-C(20、60、100 μmol·L^-1)to Al solution,the secretion of organic acids was significantly inhibited. However,the amount of organic acids secreted elevated significantly after the exposure of root apices to Al solution with heterotrimeric G-protein agonist cholera toxin(50 ng·mL^-1). These results suggested that our system for in vivo root apex culture and micro-exudates collection was a available tool for the studies on Al-induced secretion of organic acids,and organic acids were secreted from root apices of rye and wheat by anion channels in different patterns,while heterotrimeric G-protein may mediate the secretion of organic acids from root apices.

Comparison of stability of photosystem II between two Alternanthera invasive species with different invasion ability

2015/12/15 10:55:20
To reveal the invasion mechanism of an alien noxious weed-Alternanthera philoxeroides and reduce or ultimately eliminate its damage,physiological responses of A. philoxeroides and A. pungens with a relatively weak invasion capacity to photooxidative stress(induced by 10 μmol·L^-1 methyl viologen at light intensity of 100 μmol·m^-2·s^-1)were investigated and compared in the present study. Under photooxidative stress,the contents of chlorophyll,carotenoids,total phenolics and flavonoids in A. philoxeroides and A. pungens significantly decreased and leaf cell death rate correspondingly increased,but the former showed a higher tolerance to photooxidative stress,which might be related to total phenolic,flavonoid and DPPH· scavenging capacity in A. philoxeroides. This could be used to explain why A. philoxeroides had more stable photosystem,greater invasion capacity and higher damage.

Effects of forest gap size on growth and photosynthetic characteristics of Toona sinensis

2015/12/15 10:55:20
To study the growth,photosynthetic characteristics and physiological of Toona sinensis under different sizes of forest gaps,to know the response of the light intensity,we were selected three different size of forest gaps including 50 m²(Ⅰ),100 m²(Ⅱ),200 m²(Ⅲ)and opening field(CK),which were in a cypress plantation in hilly region of central Sichuan. The results showed that the plant height,stem diameter,crown area of Toona sinensis were significantly influenced by increased of gap size. The three properties in opening field were significantly lower than which in other size of forest gaps,and those properties in gap Ⅱ were the largest. The net photosynthetic rate at noon,intrinsic quantum efficiency,light saturation point,highest maximum net photosynthetic rate,photosynthetic nitrogen utilization efficiency,light use efficiency,chlorophyll a/b,leaf thickness,leaf fresh mass per unit leaf area,leaf dry mass per unit leaf area,nitrogen in per unit area,malondialdehyde,superoxide dismutase,soluble protein increased with the increase of gap size. On the contrary,the net photosynthetic rate of day,chlorophyll a,chlorophyll b,carotenoids decreased. So the photosynthetic capacity of Toona sinensis decreased with the increase of gap size,and to some extent had a positive effect on differences of photosynthetic capacity and growth. We speculated that non-stomatal limitation was the main factor that "photosynthetic siesta" and inhibited net rate. Therefore,Toona sinensis had a good adaptability to environment and Toona sinensis could keep a high photosynthetic capacity and productive forces in gap Ⅱ at the initial stage of gap formation.

Preliminary study on autointoxication and allelopathy of Panax notoginseng

2015/12/15 10:55:20
Germination and early seedling growth bioassays were used to assess the autointoxication and allelopathy potential of Panax notoginseng which served as donors and P. notoginseng,wheat(Triticum aestivum),rape(Brassica campestris)as receptors. The results were as follows:(1)the autointoxication of P. notoginseng appeared some differences in different density,but showed no obvious regularity. The exudates from seed germination of P. notoginseng inhibited seed germination rate,germination index and seedling height of rape;(2)The allelopathic effects of water extract of P. notoginseng differed among receptors. The wheat showed different promotion or inhibition which mainly manifested in the seedling fresh weight,seedling dry weight,root fresh weight,root number of wheat growth. The water extract inhibited the germination rate,promoted the seedling fresh weight,seedling dry weight,root fresh weight,root length of rape;(3)The allelopathy and autointoxication of P. notoginseng was obvious,the substances may be an important factor affecting the continuous cropping

Transformation of tobacco plants by Yali PPO-GFP fusion gene and observation of subcellular localization

2015/12/15 10:55:20
To explore the subcellular localization of Polyphenol oxidase(PPO)from Pyrus bretschneideri,the 1 779 bp cDNA of PPO gene was cloned and fused with GFP in frame to construct a binary vector pBI121-PPO-GFP. Then,the binary vector was transformed into Nicotiana tabacum by the tumefanciens-mediated method. Using confocal laser scanning microscopy,green fluorescent signals were localized in chloroplasts of the transformed Nicotiana tabacum cell. This results suggested that the Polyphenol oxidase from Pyrus bretschneideri was a chloroplast protein.

Cryopreservation of Jiangxi Yanshan red bud taro (Colocasia esculenta var. cormosus cv. Hongyayu) embryogenic callus by vitrification and its plantlet regeneration

2015/12/15 10:55:20
Using Jiangxi Yanshan red bud taro(Colocasia esculenta var. cormosus cv. Hongyayu)embryogenic calli as materials, the effects of various factors on its cryopreservation by vitrification were studied. The results showed that better preculture condition of Jiangxi Yanshan red bud taro embryogenic calli vitrification cryopreservation was 0.3 mol·L^-1 sucrose for 3 days. Better loading time was 60% PVS2 for 20 min. Better dehydration time of 100% PVS2 was 30 min at 25 ℃. Better thaw temperature was 40 ℃. Better sucrose concentration in washing media was 1.2 mol·L^-1. Better culture condition after cryopreservation was dark culture for 7 d and then transferred to the photoperiod. The average survival rate of embryogenic calli after cryopreservation by vitrification amounted to about 70%. No significant difference was observed in the morphological,physiological and cytological indexes of plantlets coming from control and cryopreserved embryogenic calli.

Tissue culture propagation technology of Aquilaria malaccensis

2015/12/15 10:55:20
Using the young shoots as explants, the methods including sterilization, induction, propagation, rooting and planting of the Aquilaria malaccensis were studied. It was indicated that the best method to sterilize was 4-5 min as 0.1% HgCl₂; 1/2 MS+6-BA 0.2 mg·L^-1+NAA 0.1 mg·L^-1+sugar 30 g·L^-1+ agar 5.8 g·L^-1 as the effective medium for adventitious shoot induction, the induction rate was 70.5%; 1/2 MS+0.1 mg·L^-16-BA+sugar 25 g·L^-1+agar 5.8 g·L^-1 as the suitable propagation medium, and the coefficient was 2.9; the medium of 1/2 MS+ sugar 30 g·L^-1+ agar 5.8 g·L^-1 made the emblings grow strong to cut; 1/2 MS+NAA 5.0+sugar 20 g·L^-1+agar 6.0 g·L^-1 as the rooting medium,the rooting plantlets would be transferred to the medium with no hormone after two days. The rooting rate was 83%. It was a little difficult to transplant, and the survival rate was only 65% in the medium mixed with peat soil and yellow mud(proportion was 2:1).

Allelopathic potential of 8-methoxy dihydrosanguinarine

2015/12/15 10:10:34
Effects of 8-methoxy dihydrosanguinarine isolated from Hypecoum leptocapum on the root growth and root hair development of Lactuca sativa seedlings were studied. The results showed that the root growth was significantly inhibited by 8-methoxy dihydrosanguinarine at the concentration of 50-200 μmol·L^-1; the inhibitory effect on root hairs increased with concentration ranging from 10 to 30 μmol·L^-1. The inhibitory effects of tested compound on the length and number of root hairs showed a dose-dependent manner. Moreover,the mitosis of apical cells could be inhibited by 8-methoxy dihydrosanguinarine at the concentrations above 50 μmol·L^-1 and revealed a positive linear regression with root NGR. Thus,the effect of 8-methoxy dihydrosanguinarine on Lactuca sativa seedlings mainly explored on the inhibition of the mitotic activity.

Physiological and ecological characteristics of Cyperus malaccensis to Cr⁶⁺ stress

2015/12/15 10:10:34
This study adopted a simulated experiment method by the means of planting Cyperus malaccensis in pots in order to study the different concentrations of Cr⁶⁺ stress effects on the plant eco-physiological characteristics and its resistance to the stress,aiming to obtain the species's toxicity mechanisms as well as its resistance to the metal,so as to provide reference to the pollution ecology of salt marsh plants. The results were as follows:(1)The biomass was inhibited drastically as the stress increased;(2)The contents of chlorophyll and the ratio of chlorophyll a and chlorophyll b reduced obviously,and there were obviously negative effects to net photosynthesis rate(Pn), intercellular CO₂ concentration(Ci), transpiration rate(Tr)and stoma conductivity(Gs);(3)The root vigor was inhibited significantly under Cr⁶⁺ stress;(4)The activity of CAT under Cr⁶⁺ stress were lower than the control group(except the concentrations of 50 mg·L^-1), the activity of SOD decreased, but the activity of POD increased all the time. The contents of MDA,proline and cytomembrane permeability increased obviously with the increasing of Cr⁶⁺concentrations. Therefore, it highlighted that the Cr⁶⁺stresses could mangle the cytomembrane system,the structures and functions of the major organelles of tea plant.

Xenia effect of triploid Siraitia grosvenorii based on fourier transform infrared spectroscopy

2015/12/15 10:10:34
The triploid female Siraitia grosvenorii of four strains were pollinated with five different diploid male flowers of S. grosvenorii,respectively. The infrared spectrums of the fruits of their offspring were measured by fourier transform infrared spectroscopy(FTIR),and the effects of male flowers on chemical component of offspring fruits were studied using principal component analysis(PCA)and clustering analysis. The results showed that there was significantly diffirent between peak height of sweet glycosides material characteristic absorption peak in infrared spectroscopy 1 050 cm^-1(wave number)among females(F302, F323 and F322)offspring fruits,which was not significantly diffirent in female F311,while five kinds of female plants pollinated male F302, F323, F322 and F311 progeny fruit in two-dimensional projection of the main component and cluster diagram sort also had a significant impact,but for different strains of fruit sorting female plants affected offspring different,thus described triploid Mangosteen sweet glycosides ingredient content material content, and overall had a more significant effect Xenia, and there is variety-specific.

Effects of different treatments on seed dormancy breaking and seedling growth of Ammodendron argenteum

2015/12/15 10:10:34
This paper studied the approriate method to break Ammodendron argenteum seed dormancy and the status of its seedling growth. The results showed that the most effective methods to break dormancy were slit testa and sand grinding. However,they could easily cause rot and death of the seeds. Use 65 ℃,80 ℃ and 95 ℃ hot water seeds treatment respectively and it was the 95 ℃ hot water treatment for 1 min that had the best breaking dormancy effect and the seeds grew well. 65 ℃ hot water treatment had the worst effect and 80 ℃ hot water could partially break the seeds dormancy. Except seeds under 80 ℃,extending hot water treatment time would increase the seeds death toll and most of the seeds under 95 ℃ died in extension period. The effects of seed soaking with liquid nitrogen treament were all significant,in which 10 min's seeds soaking was most effective in breaking domancy,nontheless,with abnormal seedling growth leading to death. Soaking with sulfuric acid for 120 min had the best effect in breaking seed dormancy. However,overtime treatment would increase the number of dead seeds and cause rot and death of a few seedling roots. In conclusion,soaking with sulfuric acid for 120 min and 95 ℃ hot water treatment for 1 min was the better method to break A. argenteum seed dormancy.

Effects of drought stress and rehydration on physiological characteristics of Ardisia corymbifera var. tuberifera seedlings

2015/12/15 10:10:34
In order to understand the physiological adaptation mechanism and recovery mechanism of karst plant under drought,this study was conducted with karst endemic medicinal plant Ardisia corymbifera. The physiological characteristics,including photosynthetic parameters,leaf relative water content,leaf membrane permeability,and osmotic regulation substances content of A. corymbifera seedlings under drought stress and re-watering conditions were studied. The results showed that under water stress,the leaf net photosynthetic rate,stomatal conductance and transpiration rate of A. corymbifera were close to zero,but the intercellular CO₂ concentration increased,suggesting that the non stomatal limitation was the main cause of the decline of photosynthetic rate. Water stress did not affect the total chlorophyll and carotenoid content per unit area of A. corymbifera,but the Chl a/b and Car/Chl was significantly lower and higher than that in control,respectively. Under water stress,the leaf relative water content,relative conductivity and MDA content of A. corymbifera increased significantly,while the membrane system was injured. Under drought stress,the proline content of A. corymbifera reduced significantly,soluble protein content did not show significant change,the soluble sugar content increased significantly,but increased marginally,these results suggested that osmotic adjustment ability of A. corymbifera to drought was weak. After re-watering,all these parameters could recover and reach the normal level. This study suggested that A. corymbifera was sensitive to drought stress and mainly employed avoidance strategies.

Photosynthetic responses to light of the four sagebrush species

2015/12/15 10:10:34
The light curves of Artemisia vestita,A. mongolica,A. japonica and A. roxburghiana were measured,and parameters of photosynthesis were calculated by non-rectangle hyperbolae function. Our results showed that A. japonica had higher light saturation point(411.0 μmol·m^-2·s^-1),higher maximum net photosynthetic rate(18.627 μmol·m^-2·s^-1)and lower light compensation point(17.867 μmol·m^-2·s^-1)than other species,revealing a higher ability to adapt highlight conditions for A. japonica. A. roxburghiana had the lowest values of dark respiration(0.353 μmol·m^-2·s^-1)and apparent quantum yield(0.038),indicating this species only maintained a low growth rate under low light conditions. Although the four species had similar trends in water use efficiency in response to increasing light densities,A. roxburghiana exhibited higher levels of WUE than those of other species,which might be reflected an adaptation to drought conditions. Furthermore,the parameters derived from light response curves exhibited a higher variation among species,especially for L_cp and R_day. Overall,those results confirmed the hypothesis that the expression of plant functional traits was closely related to the long-term adaptation of plants to environment and survival strategy.

Callus induction of Ginkgo biloba by orthogonal test

2015/12/15 10:10:34
Effects of NAA,KT,2,4-D,sucrose concentration and explants type on the callus induction of Ginkgo biloba were investigated by orthogonal test in this paper. The results showed that explants type was the most important factor among the five factors for callus induction of G. biloba,followed by KT and NAA concentration,while 2,4-D and sucrose concentration were the weakest. The best medium for callus induction of G. biloba was MS+NAA 0.5 mg·L^-1+KT 0.5 mg·L^-1+sucrose 40 g·L^-1,and stem was the best explants. With the stem as explants and with the optimal medium as the culture medium,the highest rate of callus induction in G. biloba was up to 100%.

Establishment of high frequency regeneration system for Eustoma grandiflorum cv. Excalibur Blue Picotee and determination of kanamycin sensitivity

2015/12/15 10:10:34
Eustoma grandiflorum is a popular cut flower and pot flower in the world. In this study,using leaves of sterile plantlets of E. grandiflorum cv. Excalibur Blue Picotee as explants,the effects of different concentration combinations of 6-BA and NAA on adventitious bud regeneration were investigated. The effects of different concentrations of IBA and NAA on rooting were studied,respectively. The sensitivity to kanamycin upon the regeneration of adventitious buds was determined. The results showed that the medium with MS+0.5 mg·L^-1 6-BA+0.01 mg·L^-1 NAA was the optimum medium for adventitious bud regeneration,with 91% regeneration rate. Meanwhile,the medium with 1/2MS+0.2 mg·L^-1IBA was the optimum medium for rooting,and the rooting rate reached 89%. Besides,the minimum concentration of Km inhibiting the regeneration of adventitious buds was 25 mg·L^-1. This study laid an important foundation for the research of genetic engineering on this cultivar of E. grandiflorum.

Physiological responses of Chorispora bungeana seedings in vitro to drought stress simulated by PEG-6000

YANG Ning, WANG Cheng-Liang, LI Yi-Shen, WANG Xin-Xia, CHEN Xia, NIU Tao — 2015/12/14 17:22:12
Chorispora bungeana ia a perennial herb,which grows in Qinhai-Tibet Plateau,where has a elevation of 2 600-3 700 m,and has the features of less rain,drought,low temperature,high ambient ultraviolet radiation. The special growing environment gives it the unique molecular and physiological resistance to stress response mechanism. Recently,there are many reports about the molecular and physiological resistance to the low temperature stress response mechanism. However,it has not been reported about what physiological ecology strategies it has in the process of long-term growth to adapt to drought environment. In this study,Taking C. bungeana plantlets in vitro as material,the current study was carried out to investigate the effect of drought stress on a Qinhai-Tibet Plateau plant C. bungeana with solid-liquid culture medium. There were five drought levels being set: the control(without adding PEG-6000,CK),mild drought stress(5% PEG-6000),moderate drought stress(10% and 20% PEG-6000),and severe drought stress(40% PEG-6000)and different stress duration times(6,12,24,48,72). The physiological and biochemical characters were researched. The results showed that,under drought stress and different stress duration times,the contents of proline always kept rising and was significantly higher than control during the experiment; The content of soluble protein showed an initial increase and then subsequent decrease; The content of MDA showed an initial increase and then subsequent decrease but MDA content was significantly higher than control,and chlorophyll also showed an initial increase and then subsequent decrease,and chlorophyll content was significantly lower than control under 40% PEG-6000 stress. Chlorophyll a content showed the trend of decreased significantly. Chlorophyll b content as remarkably increased under 5% PEG-6000 stress and significantely higher than control under 10%,20%,40% PEG-6000. Besides,K⁺、Na⁺、Ca²⁺、Mg²⁺ contents in leaves of C. bungeana had also obvious fluctuation. K⁺ and Ca²⁺ contents were remarkable higher than control under 5% and 10% PEG-6000 during 6 to 72 h. Na⁺ content was higher comparing with control under 5%PEG-6000 and different stress duration times,but the contents were significately decreased under the concentration and time stress. The content of Mg²⁺showed the declining trend. The study suggested that,under drought stress, C. bungeana started to dealed with drought stress by changing the osmotic regulation substances,photosynthetic system chlorophyll and ion contents. It partly revealed the physiological and ecological mechanism of C. bungeana responding to drought environment.

GmMIR160A, a class of soybean microRNA gene, negatively regulates progress of leaf senescence

2015/12/14 17:22:13
Leaf senescence is known to be a genetic programmed process regulated by the interior and/or exterior factors. Plant hormones including cytokinins,ethylene and auxin directly or indirectly regulate leaf senescence as pivotal interior factors through their redistribution and local concentration gradient by long and/or short distance transport. The molecular genetics study has shown that the cytokinins work as a repressor and the ethylene as a positive regulator respectively during the processes of leaf senescence. However,the molecular mechanism of auxin involved in regulating leaf senescence is largely unknown. The mature microRNAs are a kind of small and short double-strand RNA molecules with 21-23 bp length,which are first transcripted from mircroRNA(miRNA)genes and then processed by the special enzymes. These small RNAs are proved to be involved in many plant growth and development processes by matching their specific targets in an inaccuracy manner. However,there is no evidence that the specific microRNA could regulate plant leaf senescence in soybean so far. Soybean is a both oil crop and the classical seed-setting dependent aging species. To uncover the molecular mechanism of soybean leaf senescence should have both the important scientific significance and the profound application value in the future. In this report,we used real-time fluorescence quantitative PCR(qPCR)approach to explore the expression pattern of GmMIR160A gene during the development of the first trifoliage soybean leaves. We found that the messenger RNA of GmMIR160A was rapidly induced by both exogenous applied auxin and darkness treatment,suggesting that this gene could be both an immediate auxin-responsive gene and leaf senescence-associated gene. To further investigate the role of GmMIR160A gene during the soybean leaf development,the binary vector over-expressing GmMIR160A induced by glucocorticoid-like chemical,dexamethasone( DEX),was constructed and then transformed into wild-type soybean through Agrobacterium-mediated method with cotyledon node as the explants. Using the time order screening approaches including the antibiotic screen,the genome PCR identification and the phenotypic analysis,we finally generated four transgenic lines(Line OX-3,5,7 and 8)with stable integrated insertion T-DNA. Compared with wild types control,these transgenic plants,successful expressing the transgene showed normal morphological characteristics in respect to roots,stem,leaves,flowers,fruits and seeds but exhibited the increased chlorophyll content and higher maximum quantum efficiency(Fv/Fm)for the first trifoliage leaves during the mature stage. Moreover,GmARFs and GmCYSP1,in which the former are targets of GmMIR160 and the latter is thought as a soybean senescence marker,were down-regulated dramatically in the transgenic trifoliage leaves. Taking together,these data indicated that Gma-miR160 might negatively regulate leaf senescence by repression of its targets in soybean. This report uncovered a novel pathway that the plant hormone auxin could modify the processes of leaf senescence by regulating the transcriptional expression of microRNA gene Gma-miR160 and then repressing the messenger RNA level of auxin responsive factors GmARFs and also provided the new clues for investigating how the plant hormones control the progress of leaf senescence.

Determination of theanine and cloning of theanine synthetase gene in Dabieshan wild Camellia oleifera

2015/12/14 17:22:14
Theanine was discovered as the most abundant free amino acid in Camellia sinensis leaves with many physiological and pharmacological function. However,only limited reports have subsequently been published about theanine in the plant kingdom,for instance,mushroom,Xerocomus badius,and some theaceae plants. Theanine has been extensively studied about human nutrition for tea unique taste characteristic “umami”. Moreover,synthesize theanine has not only positive significance in plant classification,but also great economic value for the effective exploration of the plant resources. Furthermore,the mechanism of theanine synthesis in C. sinensis,separation and purification of theanine synthetase and cloning and expression of TS gene were indirectly studied. And HPLC and LC-TOF/MS were applied to determine theanine in the roots and leaves in the young and mature Dabieshan wild C. oleifera plants while molecular biology methods were used to clone theanine synthetase(TS)gene in C. oleifera; Bioinformatic analysis was carried out to analyze the gene sequence. The results revealed that the theanine content of roots in young C. oleifera plants was 0.08 mg·g^-1(fresh weight). However,theanine was not detected in leaves of young plants. It was also found there was no theanine in the roots and leaves in mature C. oleifera plants. One TS ORF(1 071 bp)was acquired from the roots in young C. oleifera plants. The homology of the cloned TS with glutamine synthetase gene(GS,AB117934)and TS (DD410896)were up to 98%. Moreover,the cloned TS showed a high similarity about 99% to GS(AB117934)and TS (DD410896)from C. sinensis at the protein levels. We found that there were 20 phosphorylation sites in the polypeptide chain after bioinformatic analysis. Bioinformatics prediction showed that the protein contained hydrophilic and winded helix domain. However,there was neither signal peptide nor transmembrane in the sequences,and the protein was non-secreted which functioned in the cell. This paper would provide theoretical basis and new approach for study of the synthesis,metabolic pathways and metabolic mechanism of theanine. Furthermore,it was beneficial to develop economical value of C. Oleifera.

Expression pattern analysis of FaGR gene of strawberry leaves under stress of different low temperatures

2015/12/14 17:22:14
In order to investigate the expression pattern of glutathione reductase from strawberry under low temperature stress,so as to further reveal its function in defensing stresses and the molecular mechanism of cold-resistant in plants. We treated strawberry plants(Fragaria&#215;ananassa‘Toyonaka')with different low temperatures including 0,2,4,6,8 and 10 ℃ in artificial chamber for 24 h respectively,and we also treated plants with 25 ℃ for 24 h as control. Subsequently,total RNA samples were extracted from the treated fresh young leaves,and cDNA first strand for PCR reactions was synthesized using these RNA samples. When all cDNA samples were done,we also chose two methods including semi-quantitative PCR and real-time quantitative PCR to detect the expression differences of FaGR between different low temperature treatments. We chose two different methods to determine the expression of FaGR,on one hand to make sure the differences of expression between samples were exactly due to the different temperatures,not because of the different methods; on the other hand,it was expected to see whether semi-quantitative PCR was reliable as we thought for gene expression research. At last we combined the results came from two methods and analyzed the expression differences of FaGR gene between samples treated with different temperatures. The results were described as below: the result of semi-quantitative PCR was generally consistent with that of real-time quantitative PCR,both of them showed that the relative expression of FaGR was greatly modified in diverse grades by different low temperatures. Furthermore,the relative expression levels of FaGR under low temperature at 8 and 10 ℃ increased compared to the control which was at 25 ℃,and the highest relative expression level of FaGR was detected at 8 ℃. However,thereafter,the relative expression level at 6 ℃ dropped dramatically to a level which was slightly lower than the control at 25 ℃. When the temperature was below 6 ℃,the relative expression level of FaGR decreased accompanied with the decreasing of temperatures,each of the expression level at temperature below 6 ℃ was lower than that at 25 ℃. The lowest relative expression level was detected at 0 ℃,which was almost only the half amount of the maximum expression at 8 ℃. These results supported that semi-quantitative PCR was exactly reliable and it could be widely used for gene expression research. Also,these results indicated that FaGR gene was definitely induced by chilling stress but within a certain range of temperature,and the degree of changes under different low temperatures was also different. In a certain range,FaGR was up-regulated by low temperature,however it was slightly suppressed by low temperature out of this range,and 6 ℃ was showed as the critical value in this research. What's meaningful,these results would provide a scientific basis for the further regulation of the gene expression,as well as some basic data to improve the ability of plants to resistant stresses.

Role of the alternative respiratory pathway in mediating the cell death induced by NaCl stress on tobacco suspension cells

2016/5/4 16:06:55
Soil salinity stress is the one of the most widespread environmental stresses that growing plants have to frequently face. The effects of salinity stress on plants are generally associated with the disruption of metabolism. In particular, high level of salt stress can cause cell death. This seriously limits the development, growth, survival, and productivity of plants. The alternative respiratory pathway is a plant-specific respiratory pathway, which does not exist in animals. By using tobacco suspension cells, the present work investigated whether the alternative respiratory pathway could play a role in mediating the high-salt-induced death of plant cells. And, the possible mechanism for this function of the alternative respiratory pathway was also studied. The results showed that with the increase of the concentrations of NaCl, the levels of cell death of tobacco suspension cells increased, and the increased levels of cell death were followed with the increases of the capacity of the alternative respiratory pathway. Similar to the treatment with NaCl, the treatment with exogenous hydrogen peroxide(H₂O₂)also induced an increase in the level of cell death. The relationship among the alternative respiratory pathway, cell death, and H₂O₂ production was studied in the tobacco suspension cell subjected to 200 mmol·L^-1 NaCl. The treatment with 200 mmol·L^-1 NaCl caused an obvious increase of cell death and a significant enhancement of the H₂O₂ production. The 200 mmol·L^-1 NaCl-stressed cells pretreated with salicylhydroxamic acid, a specific inhibitor of the alternative respiratory pathway, exhibited higher levels of cell death and H₂O₂ production than the tobacco suspension cells subjected to 200 mmol·L^-1 NaCl stress alone. These results suggest that high salt stress can induce an increase of the alternative respiratory pathway, which plays a role in alleviating cell death under high salt stress. And, this function of the alternative respiratory pathway may relate to its ability to limit the generation of H₂O₂.

Water absorption properties of Salvia coccinea seed and its germination characteristics under drought stress

2016/5/4 16:06:55
Producing mucilage is a survival strategy of plants that formed in the long-term process of their environment adaptation, and the seeds producing mucilage are called mucilaginous seeds or mucus propagule. Studies show that mucilage has important ecological significance for the dispersal, settlement, viability improvement, germination, survival of seedlings and even the resistance toward the poisoning of toxic chemicals of seeds. Salvia coccinea is a perennial herb(Lamiaceae: Salvia)originating in the tropical regions of the American continent. Due to its high medicinal and ornamental value, S. coccinea is widely cultivated throughout the world. In order to understand the water absorption properties of mucilage on the seeds of S. coccinea and germination characteristics of S. coccinea seed under drought stress. In the present study, seeds of S. coccinea were studied on the morphology and water absorption of seed coat mucilage and seed germination under the drought stress which was created with different concentrations of PEG. The study methods were light micro-scope, scanning electron microscopy observation and seed germination test. The results showed that seeds of S. coccinea were spherical; 1 000-grains weight of the seeds was(1.611±0.0084)g; seeds without mucilage were able to absorb an amount of water of 3 times of their own dry weights; seeds were 25 times of their own dry weights; mucilage was 122 times of its own dry weight. The results indicated that the weights of mucilaginous seeds, non-mucilaginous seeds and mucilage layer increased with the extension of time, but the process of dehydration was much longer than that of absorption. Water absorption of the seeds reached saturation 2 h after watering, and the water saturated seeds were dehydrated to their original dry weights in 36 h after being exposed to air at room temperature. PEG of different concentrations had various impacts on the seed germination of S. coccinea, and the germination potential decreased significantly with the increase of PEG concentration. Seed germination rate was as high as(90.00±8.66)% under 5% PEG and as low as(76.67±10.41)% under 20% PEG. In conclusion, S. coccinea seeds are characterized by fast germination. Mucilage plays a decisive role in the water absorption process of seeds and ensures sufficient moisture supply for seed germination in a short time.

Interactive effects of CO₂concentration and temperature on physiological parameters of Saussurea kansuensis

2016/5/4 16:06:55
Saussurea kansuensis, as a kind of the genus Saussurea of Asteraceae, is an endemic species in the eastern of Qinghai-Tibet Plateau. Our objective was to study the effects of elevated CO₂concentration and increasing temperature on physiological parameters of S. kansuensis by using CO₂ artificial climate box(PRX-250C-CO₂, Shang Hai). The results showed that elevated CO₂concentration and increasing temperature had significant interactive effects on physiological parameters of S. kansuensis. The chlorophyll, soluble sugar and soluble protein contents of S. kansuensis reached the maximum, while the minimum content of MDA and superoxide anion was observed when the CO₂ concentration was 550 μmol·mol^-1. The chlorophyll, soluble sugar and soluble protein contents increased under higher temperature, while the contents MDA and superoxide anion decreased. At CO₂concentration of 550 μmol·mol^-1, elevated temperature could significantly enhance the amount of chlorophyll, soluble sugar and soluble protein, and reduce MDA and superoxide anion free radical contents. The results indicated that the increasing CO₂concentration and temperature could promote the growth of S. kansuensis to a certain degree.

Seed coat obstacle and endogenous inhibitory activity of various parts of Handeliodendron bodinieri seed

2016/5/4 16:06:55
Handeliodendron bodinieri is the relict of rare plant single taxa in China, due to man-made destruction, the special growth environment and the influence of its owe characteristics, with fewer resources, Handeliodendron is listed as national key protection wild treasure and endangered plants. In order to study the effects of rare and endangered H. bodinieri testa on seed germination, through dealing with different conditions(with the seed coat, concentrated sulfuric acid processed testa, the complete removal of the seed coat, revealing only radicle)of seed germination, seed coat permeable skin and the perivarp, aril, seed coat and kernel of Handeliodendron in different concentrations(0, 3.125, 6.25, 12.5 and 25 mg/mL)of methanol extracts of planting cabbage seed germination and seedling growth were studied. The results were as follows:(1)H. bodinieri tasta had good permeability, water absorption without affecting before seed germination. But testa mechanical obstruction, easy to mold had great impact on seed germination and growth.(2)Methanol extracts from fruit shells, arils, seed coats and kernels of H. bodinieri seeds affected the germination and growing of Chinese cabbage seeds and especially greatly affected the growth of radicle. The intensity of inhibition activity was: kernel>fruit shell>aril>seed coat. And the inhibition activity enhanced with the increasing of concentration. The research results revealed the reasons of germination difficulty and the low germination rate and as well to establish the foundation for the artificial propagation and protection and use of H. bodinieri.

Expression analysis of MVA and MEP metabolic pathways genes in latex and suspension cells of Hevea brasiliensis

2016/5/4 16:06:55
The MVA and MEP metabolic pathways are two important plant isoprenoid metabolic pathways in plants. The expression of genes respectively in MVA and MEP secondary metabolic pathways were analyzed in the latex and suspension cells from anther-derived callus of Hevea brasiliensis by using qRT-PCR technology. In addition, expression changes of HbAACT3, HbHMGR4, HbHMGR5, HbDXS2, HbDXR and HbSQS1 genes were further analyzed in the suspension cells under COR treatment. The results demonstrated that expressions of HbAACT1,HbAACT2,HbHMGS1,HbHMGS2,HbHMGR1,HbHMGR3,HbMVK,HbPMK,HbMVD1,HbMVD2 in MVA metabolic pathway and HbIPPI1 and HbFDPS1 genes involved in IPP utilization were relatively higher in latex than that in suspension cells, while HbDXS1,HbDXS2,HbDXR,HbCMS1,HbCMS2,HbCMK,HbMCS1,HbMCS2,HbHDS and HbHDR in MEP metabolic pathway and HbSQS1 were relatively higher in suspension cells than that in latex. Moreover, HbHMGR5, HbHMGR4, HbSQS1, HbDXS2 and HbDXR genes were induced highly or to some degree in suspension cells by COR application. This study lays a foundation for further utilization of suspension cells to analyze secondary metabolism regulation as well as to produce bioactive compounds from anther-derived callus of Hevea brasiliensis in the future.

Cloning and expression of the GGPP synthase gene from the paclitaxel-producing endophytic fungus (Penicillium aurantiogriseum)in Corylus avellana

2016/5/4 16:06:55
Geranylgeranyl diphosphate synthase is one of the key enzymes in taxol synthesis pathway in paclitaxel-producing endophytic fungus. To study taxol synthesis in paclitaxel-producing endophytic fungus, we need to identify the existence of GGPP synthase in the first place. With RT-PCR method, we cloned the ORF fragment of PaGGPPS (GenBank accession number: KM881430)from the paclitaxel-producing endophytic fungus Penicillium aurantiogriseum in Corylus avellana. The bioinformatics methods were employed to analyze and predict the composition of nucleic acid and amino acid sequence of this gene. The results showed that the length of ORF was 1 113 bp, the molecular weight of the protein was 40.98 kD, and the theoretical isoelectric point was 6.168, which suggested that PaGGPPS protein was acidic. The hydrophilicity analysis found that PaGGPPS was a hydrophilic protien. The α-helix was the dominant secondary structure constructional element of the protein which contained one tpolyprenyl_synt function domain. The identity alignment of GGPPS nucleic acid sequences of taxol producing endophytic fungi in hazelnut and other species showed that the identity were highest between Penicillium roqueforti, Aspergillus clavatus and Neosartorya fischeri, respectively 94%, 76% and 76%. The homologous alignment of Amino acids showed that GGPPS genes were divided into two groups. The GGPPS genes from animal, fungal and yeast were clustered in one group, the GGPPS genes from plant were in the other group. The homologous alignment of nucleic acid sequences showed that the homologous were the highest between GGPPSs in paclitaxel-producing endophytic fungus(Penicillium aurantiogriseum)in Corylus avellana and in Penicillium roqueforti, and GGPPSs in paclitaxel-producing endophytic fungus and plant have the longest distance in the phylogenetic tree. After the bioinformatics analysis, we constructed a pET30a prokaryotic expression vector of PaGGPPS. The protein of PaGGPPS was successfully expressed in the soluble state in Escherichia coli. All the experiments we had done can provide some information for the further research.

Discovery and identification of homology gene to Rpi-blb2 in tomato(Solanum lycopersicum)

WANG Jun-Sheng, HU Li-Zong, LIU Hong-Zhan, WANG Yong-Li, CHEN Long — 2016/5/4 16:06:56
Late blight is the most destructive disease in tomato(Solanum lycopersicum)cultivation worldwide and will result in large economic losses. As the variation and evolution of original races, mining new resisitance gene and improving the genetic resistance to late blight is a major issue in breeding new varieties of tomato, because of the new races will overcome the resistance gene-mediated genetic resistance from commercial varieties. Rpi-blb2, a potato late blight resistance protein as seed sequence, has 11 homology sequences from tomato protein sequences databank in NCBI and has a similarity to seed sequence range from 78%-83%. The above data were obtained by blast-protein way in this study. Then the gene structure, gene location, conserved domain feature of 11 protein sequences, and homology sequences of Rpi-blb2 belong to disease resistance protein sup-family of tomato and their evolution relationship were revealed and discussed. The results showed that ten of eleven distributed in the sixth chromosome, one the fifth chromosome; ten sequences distributed the sixth chromosome showed two gene clusters, containing two and three gene locus in the sixth chromosome, respectively. They were co-orthologous for Rpi-blb2 protein, but not paralogous each other, the most of them located in cytoplasm. According to the difference in structure domain and gene location, they were divided into three categories. The first category, a total of four members, contains DUF3542 and NB-ARC conser domain; The second category, a total of six members, like potatoes Rpi-blb2 protein, containing only NB-ARC conserved domain, the NB-ARC domain is located in the middle of these two kinds of protein sequences; The third category(only contains XP_004239406. 1)also has DUF3542 and NB-ARC domain and located on chromosome 5, but the non-conserved sequence at the ends of the domain is shorter. The members of the first two categories have one or two introns in gene structure, the third category does not contain introns. The study provides theory basis for breeding tomato disease resistance cultivar by molecular biology techniques.

Bioinformatics analysis of DXS gene from six tropical plants including oil palm(Elaeis guineensis Jacq.)

2016/5/4 16:06:56
The paper unraveled structure characteristics, function features and phylogenetic relationships among species through analyzing nucleic acid and protein sequences of key gene DXS controlling isoprenoids synthesis of tropical plants as oil palm(Elaeis guineensis Jacq.)by using bioinformatics tools. Isoprenoids like carotenoids and sterols play an important role in life of plant, and have important significance for the protection of human health. Tropical plants as oil palm contains abundant isoprenoids like carotenes and vitamin E, MEP is one of the important pathway for isoprenoids biosynthesis. DXS as the first rate limiting enzyme in MEP pathway, its functions were highly conserved in diverse tropical plants. In order to understand the structure and function characteristics of DXS from tropical plants as oil palm, DXS genes nucleic acid and protein sequences from different plants including oil palm were analyzed such as physicochemical property, protein structure, and conserved domain by the tools of bioinformatics in this study. The results showed that the start codon of DXS gene was ATG, but termination codon was divided into TAG, TAA and TGA; DXS protein belonged to a hydrophilic protein without a signal peptide; It might played a role in the chloroplast stroma as a transport protein; No transmembrane domain was found, there were 36 phosphorylation sites, including 17 serine, 11 threonine and 8 tyrosine sites respectively; Random coil and alpha helix were the major structural elements of secondary structure; It had DXS enzyme character, thiamin diphosphate(TPP)binding sites and PLN02582 conserved multi-domains with tertiary structure analysis; Domains of DXS protein from different plant, were very conservative, and could be used as an important basis for determining genetic relationships among species. The results would provide further information for structure and functional analysis of DXS, and lay the foundation for molecular mechanism research and genetic improvement of quality traits for tropical plants including oil palm.

Optimal medium for pollen germination of Jatropha curcas in vitro using response surface methodology

2016/5/4 16:06:56
Jatropha curcas is likely to become one of the key energy crops in the world because of high oil content in seed which can be refined into high quality biodiesel. J. curcas widely distributes and cultivates in many tropics and sub-tropics areas of China. Due to inadequate pollination and very low productivity, J. curcas was a wild or semi-wild tree for a long time. Effective pollination is a prerequisite for fruit-set and seed-set of J. curcas. Information on the most suitable ingredient concentration ratio of B-K medium which was widely used to pollen culture in vitro for pollen germination and inactions among ingredients in vitro is required to increase rate of hybrid seed set and seed production. Gathered mature pollen grains when petals opened and the anther just began scattering pollen in primary flowering time to do pollen germination test. Ingredient concentration ratio was optimized by response surface methodology(RSM)based on the Box-Behnken design(BBD). The primary variables chosen were sucrose, H₃BO₃, Ca(NO₃)₂·4H₂O and KNO₃. A total number of 30 runs were tested. We constructed a response surface model between the pollen germination rates and four ingredients for the culture medium in vitro by R 3.0 software. The effect order of 4 factors to influence germination rate were: sucrose > H₃BO₃ >Ca(NO₃)₂·4H₂O>KNO₃. There were significant interactions of pollen germination between sucrose and H₃BO₃, sucrose and Ca(NO₃)₂·4H₂O, sucrose and KNO₃. The optimal ingredient concentration ratio which supported the germination of pollen and growth of pollen tubes was 13.77% sucrose + 32.14 mg·L^-1 H₃BO₃+ 22.21 mg·L^-1 Ca(NO₃)₂·4H₂O + 19.95 mg·L^-1 KNO₃ + 200 mg·L^-1 Mg(SO₄)₂·4H₂O. Under this condition, the germination rate theoretically achieved 99.73%. The verification test were made corresponding to the optimal ingredient concentration ratio, the results of pollen germination experiments showed that pollen germination rates of 98.97%, the proportion matched theoretical response value well. It can also verify the effectiveness of the BBD-based response surface model and analysis technique in obtaining optimal ingredient concentration ratio for pollen germination.

Influence of exogenously applied methyl jasmonate on soilborne pathogen, Cylindrocarpon destructans in vitro

2016/5/4 16:06:56
Ginseng(Panax ginseng)is a traditional medicinal plant in China, and the dried root is highly valued for its medicinal properties and is widely used in Chinese traditional medicine. Cylindrocarpon root rot of ginseng caused by Cylindrocarpon destructans is the main disease, which is difficult to be eliminated from soil, leading to serious crop yield losses and limited the re-use of fields for successive ginseng crops. Methyl jasmonate(MeJA), which is one of the plant lipid derivatives, considered as a signaling substance during plant-microbe interactions. It is related to plant resistance to biotic and abiotic stress, can be involved in plant response to pathogens and other adversity stress and signal transmission, and can be used to induce plant disease resistance reaction. The objective of this study was to test the effects of MeJA on C. destructans and to find the possible relationships between MeJA and pathogenic factors of C. destructans. To assess the influence of MeJA on soil pathogens, the direct effects of artificially applied MeJA on C. destructans were evaluated, including the effects on colony growth, spore germination, biomass and phytopathogenic enzyme activities of this pathogen. The results showed that MeJA strongly inhibited its growth and spore germination, whereas it stimulated phytopathogenic enzyme activities. The colony diameter of C. destructans on PDA decreased from(8.23±0.15)cm(control)to(0.71±0.00)cm(800 μg·mL^-1). At the highest concentration(800 μg·mL^-1)of MeJA, the colony diameter of C. destructans had almost totally inhibited, but the diameter had no difference compared with the untreated control at lower concentrations(1-50 μg·mL^-1 MeJA). The biomass reduced by 65.3%-100% and the percent of spore germination decreased by 100% at concentrations higher than 400 μg·mL^-1. Potent suppression of the growth of germ-tubes was observed at different concentrations, especially at(400-800)μg·mL^-1, where the growth of germ-tubes was inhibited completely. However, activities of phytopathogenic enzymes(pectinase, cellulase, amylase, and protease)were greatly stimulated by MeJA at higher concentrations(MeJA>200 μg·mL^-1), while the activity of protease was little changed. The activity of pectinase was high up to(0.61±0.05)U·mL^-1·min^-1 at the concentration of 800 μg·mL^-1. The activity of cellulase was stimulated at high concentrations of MeJA in liquid culture, while it was suppressed at low concentrations(1-50 μg·mL^-1). The activity of cellulase was(0.31±0.02)μmol·min^-1 at 800 μg·mL^-1 of MeJA. Amylase activity substantial increased at concentrations of(200-800)μg·mL^-1, which was(0.45±0.02)μmol·min^-1 at the concentration of 800 μg·mL^-1. It was concluded that MeJA greatly inhibited C. destructans growth and spore germination, but stimulated activities of hydrolytic enzymes of C. destructans at higher doses. There might be different mechanisms presented for the effects of MeJA on hyphal growth and virulence factors of C. destructans. The critical concentration of MeJA inhibitory effects on C. destructans was 200 μg·mL^-1. The research lays a foundation for the subsequent experiment using MeJA to induct disease resistance. This is of help to understand the mechanism of Cylindrocarpon root rot of ginseng and to control pathogen in practice.

Fresh-keeping of Toona sinensis sprouts with tea polyphenol combined with chitosan solution

2016/5/4 16:06:56
The fresh-keeping effects of tea polyphenols(TP)combined with chitosan solution on the sprouts of Toona sinensis were studied. The sprouts of T. sinensis were treated with TP combined with chitosan of different concentrations and storaged at 4(±1)℃, the fresh-keeping effects were studied by determination of sense assess, the physiological and biochemical index, and the rot rate of the sprouts. The results showed that certain concentration of TP combined with chitosan solution could maintain the sensory quality, and reduce the biological senescence, among which 0.3% TP with 0.5% chitosan had significant effects on the fresh-keeping of T. sinensis. The weight loss rate of the sprouts treated with 0.3% TP-0.5% chitosan was only 1.52% on the 12th day of preservation, vitamin C retention rate was 63.08%, chlorophyll loss rate was 20.67%. The solution of 0.3% TP with 0.5% chitosan could effectively reduce the rot rate and defoliation rate, the intact rate was 84% and the defoliation rate was 8% after preservation for 21 d. However, the solution of 1% TP aggravated the corruption of the leaf, covered the original fragrance of the sprouts of T. sinens, and even caused the browning. As a result, the solution of 0.3% TP with 0.5% chitosan has a good application value on the fresh-keeping of sprouts of T. sinens.

Physiological mechanism of cassava South China 8 tissue culture seedling under salt stress

2016/12/26 11:22:52
The cassava(Manihot esculenta)South China 8(SC8)tissue culture seedlings were conducted to investigate the physiological mechanism in response to salt stress. Effects of MS medium treatment contained NaCl(0, 5, 20, 35, 50 mmol·L^-1and R50 mmol·L^-1)on the growth and physiological indexes activities of Cassava SC8 tissue culture seedlings were evaluated. The physiological indexes include chlorophyll, hydrogen peroxide(H₂O₂), malonaldehyde(MDA)content, and the activities of superoxide dismutases(SOD), catalases(CAT), peroxidase(POD), ascorbate peroxidase(APX). The results of physiological showed that the growth of SC8 tissue culture seedlings did not change significantly after NaCl content(≤20 mmol·L^-1)stress for 60 d, but high content(≥35 mmol·L^-1)of the stress had inhibited SC8 tissue culture seedlings growth. The growth of SC8 tissue culture seedlings could be recovered at NaCl concent(50 mmol·L^-1)stress for 30 d after they were transferred to the normal MS medium of 30 d. Chlorophyll and MDA contents were accumulated at NaCl content(≤35 mmol·L^-1)stress, but which were not accumulated at NaCl content(≥50 mmol·L^-1)stress. H₂O₂content had no obvious change at NaCl content stress which it was compared with control. Activities of antioxidant enzymes including POD, CAT and APX were increased at NaCl stress, whereas the activities of SOD, CAT and APX were decreased at higher NaCl stress. The results of real-time quantitative PCR showed that the expression level of SOD, CAT, POD and APX were higher than control at NaCl content(≥50 mmol·L^-1)stress. The results indicated that salt stress for a short period of time would not cause devastating damage to cassava, which could improve the salt tolerance of cassava by regulating the activity of physiological indexes.

Effects of IBA concentration and cutting time on rooting ofRhododendron kiangsiense and Rh. liliiflorum cuttings

2016/12/26 11:22:52
To find out the best IBA concentration and cutting time for rooting of Rhododendron subsect. Maddenia(Hutch.)Sleumer cuttings, we studied the effects of four IBA concentrations and four cutting time on rooting of cuttings of Rhododendron kiangsiense and Rh. liliiflorum, using soil + sand(1:1)and peat + perlite + vermiculite(3:1:1)as rooting media. The results showed that IBA concentration had significant effects on most parameters except for retention number of old leaves. Cuttings treated with 100 mg·L^-1 IBA had the highest rooting rate and length of new shoot and the lowest rotting rate, with good performance of other parameters, and that was the optimum IBA concentration for rooting. Followed by 50 mg·L^-1 IBA, cuttings had the highest root width and new shoot rate but the lowest number of adventitious root. The solution of 200 mg·L^-1 IBA could promote the development of root, but it gave a low rooting rate and significantly inhibited the development of new shoot. The contrast had the worst effects on rooting. Cutting time had significant effects on all rooting parameters, and early spring(April 18th)with hardwood cuttings was the optimum cutting time, which gave excellent effect on most parameters except retention number of old leave, followed by autumn(October 19th)with cuttings transforming from semi-hardwood to hardwood. And summer with softwood cuttings(June 21st)or semi-hardwood cuttings(August 16th)had the worst effects, so that was not suitable for cutting propagation. Species and rooting medium also had significant effects on rooting parameters, such as Rh. liliiflorum had better rooting capacity than Rh. kiangsiense, and peat + perlite + vermiculite(3:1:1)was better than soil + sand(1:1). This study found for the first time that using hardwood cuttings in early spring before new shoots sprouting could significantly promote rooting of these two species cuttings, and that provides the information for cutting propagation of Rhododendron subsect. Maddenia species.

Cloning and expression analysis of 1-deoxy-D-xylulose 5-phosphate synthase gene in Isodon rubescens

2016/12/26 11:22:52
1-Deoxy-D-xylulose-5-phosphate synthase(DXS)catalyses the first committed step of the 2C-methyl-D-erythritol-4-phosphate(MEP)pathway, and its expression level affects the contents of terpenoid. This study was aimed to clone the 1-deoxy-D-xylulose 5-phosphate synthase(DXS)gene from Isodon rubescens and analyze the bioinformatics and expression of the gene.The primers were designed according to the transcript sequence of IrDXS from I. rubescens transcriptome database. The physical and chemical characteristics, signal peptide, subcellular localization, secondary structure of IrDXS protein were analyzed and predicted by the tools of bioinformatics. In this work, a DXS cDNA(IrDXS, KT831764)was isolated from I. rubescens. The full-length cDNA of IrDXS encoded 722 amino acid residues with a predicted molecular mass of 77.7 KD. Sequence alignment showed that IrDXS had high homology to known DXS proteins from other plant species and contained the conserved N-terminal plastid transit peptide, the N-terminal thiamine binding domain and pyridine binding DRAG domain. Phylogenetic analysis indicated that IrDXS belonged to the plant DXS2 cluster. Quantitative real-time PCR analysis showed that the expression of IrDXS was tissue-specific, and accumulation of transcripts was greater in roots and leaves. This study provides valuable information for future experiments on the molecular mechanisms underlying the isoprenoid biosynthesis in I. rubescens.

Effects of endogenous NO and ROS generation in alfalfa under Cd stress

2016/12/26 11:22:52
Seedlings of alfalfa variety Medicago sativa cv. Gannong No. 3 were grown in nutrient solution with the Cd concentration between 0 to 2.0 mmol·L^-1. The generation patterns of endogenous nitric oxide(NO)and reactive oxygen species(ROS)in roots, stems and leaves were studied, and the changes of root activity was determined. The results showed that NO content in seedlings presented a tendency of increasing first and decreasing afterwards, finally recovering normal with increasing concentration of Cd from 0 to 2.0 mmol·L^-1. The NO content was influenced significantly by nitric oxide synthase(NOS)and nitrate reductase(NR)in stems, while NOS, nitrite ions(NO₂^-)and NR determined the NO content in leaves. Similarly, NOS and NO₂^- had great relevance with NO content in roots. The increase of Cd concentration in solution leaded to the significantly rising of content of hydrogen peroxide(H₂O₂), malondialdehyde(MDA), superoxide anion(O^-₂·)and relative electric conductivity(REC)in seedlings, indicating that the stress of high Cd concentration made the ROS vast accumulated, the cell membrane damaged, and cytoplasm outflowed, further caused membrane lipid peroxidation. Besides, there was no significant correlation between NO and ROS due to their different mechanisms of generation.

Relationship between sucrose's synthesis, distribution and the starch accumulation in the root tuber of cassava

2016/12/26 11:22:52
Two cassava cultivars with different starch contents were taken as materials in the study, and were Fuxuan 01 and SC124. The relationship between sucrose's synthesis, distribution and the starch accumulation in the root tuber was studied. Leaf, stem and root sucrose accumulation and starch accumulation in the root tuber of different cassava cultivars were determined, and their relations were analyzed in the different growth periods, respectively. The results showed that the content of sucrose in Fuxuan 01's leaf and stem washigher than that of SC124 in all periods, and the accumulation of sucrose in root was higher than that of SC124 before initial stage of root tuber thickening, but lower after initial stage of root tuber thickening. In the whole growth period, the starch content and starch synthesis rate in the root tuber of Fuxuan 01 were higher than that of SC124. Root's sucrose content was significantly declined with the growth period delayed. The change trend of sucrose content in leaf and stem was opposite to that of starchsynthesis rate, which indicated that sucrose content in leaf and stem decreased while the root starch accumulation rate increased, and the sucrose content in leaf and stem increased while the rate of starch synthesis reduced obviously. The root was the part with the highest content of sucrose, then followed by the stem, and the part with the lowest content of sucrose was leaf in the early root formation period. But the trend was opposite in the mature period, it was the leaf part with highest content of sucrose, then followed by the stem, and the part with lowest content of sucrose was root. The results of correlation analysis showed that the correlation between the sucrose content in leaf, stem and the starch accumulation in root tuber of cassava was positively related. The relation was significant between the starch accumulation in root tuber and the sucrose content in leaf but not in stem. The correlation between the starch synthesis in root tuber and the sucrose content of cassava was significantly negatively related. The relationship between sucrose's transportation, distribution and the root's starch accumulation in cassava was closely related, and the abilities of synthesizing sucrose in leaf and utilizing sucrose in root tuber play key roles in the process of starch accumulation. This study provides the information for the production and breeding of cassava.

Physiological response of transgenic tobacco to drought stress simulated by PEG6000

CHEN Xia, YANG Pen-Jun, ZHANG Xu-Qiang, YANG Ning — 2016/12/26 11:22:52
Chorispora bungeana is a rare alpine subnival plant species, the extremely harsh living environment makes it become a new plant for study the responses of drought stress. In this study we used CbPLDα, CbPLDβ transgenic tobacco plants as material to find the mechanism of plants response drought stress, and we also detected the osmolytes content and protective enzymes activity in CbPLDα, CbPLDβ transgenic tobacco plants, using the ways of the drought stress simulated by PEG6000. The results showed that proline, solube sugar and soluble protein involved in strengthening resistance of CbPLDα, CbPLDβ transgenic tobacco to drought stress simulated by PEG6000,and in different concentrations of PEG6000 simulate drought stress, CbPLDα, CbPLDβ transgenic tobacco proline, soluble sugar and soluble protein content were always significantly higher than that of wild type tobacco plants(P<0.05). This indicated that osmotic adjustment ability of drought stress in CbPLDα and CbPLDβ transgenic tobacco was stronger than the wild type tobacco plants. In the protective enzymes, Superoxide dismutase(SOD)and Peroxidase(POD)have significant complementary effects on mitigating the injury of membrane lipid peroxidation of transgenic tobacco; Catalase(CAT)and Ascorbate peroxidase(APX)played major roles in the mechanism of removal of hydrogen peroxide. SOD,POD,CAT and APX had higher activity in CbPLDα, CbPLDβ transgenic tobaccos than in wild tobacco plants(P<0.05). All of these results indicated that protective enzymes had important biological function in protecting transgenic tobacco from drought stress simulate by PEG6000. It partly revealed the physiological and ecological mechanisms of the CbPLDα, CbPLDβ genes responding to drought environment in C. bungeana. The results showed that both CbPLDα and CbPLDβ transgenic tobaccos involved in the regulation pathway of membrane stability, osmotic adjustment and the regulation of anti-oxidative system. CbPLDα and CbPLDβ genes can provide powerful genetic resources for improving drought resistance of plants. The results strengthen the PLD for functional studies, supplementary plant resistance to drought stress and drought resistance theory breeding germplasm development and utilization, has important theoretical value and practical significance.

Tandem coexpression of CaXMT and mutants of TCS1 and analysis of enzyme activity in vitro

2016/12/26 11:22:52
Caffeine and theobromine as the major component of alkaloids in tea, while caffeine is the important taste compound. As the application of caffeine increased extensively in the fields of food and medicine, caffeine biosynthesis becomes a new hotspot. Nowadays, exploring the method of caffeine biosynthesis is of great significance, when caffeine synthesis mainly relies on chemical synthesis on the market. This research concatenated coffee xanthosine methyltransferase gene(CaXMT)and four mutants of tea caffeine synthase gene(TCS1)respectively in the same expression vector pMAL-c5X, then induced coexpression of the fusion protein, which was analyzed by SDS-PAGE gel electrophoresis, and chromatography were added into the crude enzyme solution by sonication methionine. At last, in vitro enzymatic reaction products were detected by high performance liquid. The results showed that only theobromine generated in the products of pMAL-CaXMT-TM2/3/4 without caffeine. This research provides the information for establishing tandem gene coexpression vector which was used for the biosynthesis of caffeine and theobromine and new ideas to study on caffeine biosynthesis.

Screening of qRT-PCR reference genes for Chenopodium album and C. glaucum of Chenopodiaceae

2016/12/26 11:22:53
Selection of suitable reference gene is a critical step in real-time quantitative PCR(qRT-PCR)analysis. So far, reports on reference gene screening on stress-tolerant gene expression of Chenopodiaceae species are limited. In the present study, by using three reference-gene-analysis softwares-GeNorm, NormFinder, BestKeeper, the stability of three commonly used candidate reference genes β-ACTIN, β-TUBULIN and GAPDH of Chenopodium album and C. glaucum under NaCl and PEG treatments were compared. The results showed that under NaCl and PEG stresses,GAPDH showed stable both in C. album and C. glaucum. The results also demonstated that the same reference genes expressed stable in the same family. β-ACTIN expressed stable in C. album, while β-TUBULIN expressed stable in C. glaucum. We also studied the plants of different gera in the same family and the same stress of the stability of the expression of reference genes. The results suggested that same reference gene expressed stable in the same species. For the same stress in these two species, the results showed that selection of the most stable reference gene depended on the experimental materials and the conditions. The analysis results of the three candidate reference genes were based on the three analysis softwares. In our study, we determined the ralatively stable reference genes in C. album and C. glaucum. This study provides the reference for the further study on qRT-PCR analysis of stress-relevant gene expression in Chenopodiaceae species.

Changes of antioxidant enzyme and ultrastructure in recalcitrant seeds of Panax notoginseng during after-ripening process

2016/12/26 11:22:53
The changes in ultrastructure, activity of antioxidant enzyme and expression of antioxidant enzyme were examined in 3-year-old Panax notoginseng seeds during the after-ripening process. The objective of the present study was to analyze the desiccation sensitivity in recalcitrant seeds of P. notoginseng at the physiological, ultrastructural and transcriptomic levels. The results showed that during after-ripening process of 40 d, the activity of SOD, CAT, APX and GR in seeds increased at first and then reduced, and POD activity gradually increased. At the end of after-ripening period of 40 d, FPKM values of gene related to antioxidant enzymes(SOD, POD, CAT and APX)were 28, 13, 356 and 105, respectively, and all at a high. At the same time, intact cell structure was observed, and mature embryo was morphologically completed. However, the amount of MDA reached a peak value. Summarily, the peroxidations of membrane lipid caused cell membrane degradation and led to cell function loss and even cell death; meanwhile, antioxidant enzymes could not effectively resist the oxidative injury with the prolonged time of after-ripening, which may be the vital reasons for desiccation sensitivity in recalcitrant seeds of P. notoginseng.

Preservation of medicine and edible health fruits of Kadsura coccinea

2016/12/26 11:22:53
We took the fruits of Kadsura coccinea as experiment material, with glycerol, starch, gelatin, agar, sodium metabisulfite according to three different proportions made out to different antistaling agent to treat these fruits,then put them in the average temperature of 11 ℃,average humidity of 85% in the refrigeratorfresh layer to keep fresh preservation. Prepared with chitosan into four different concentrations of mass fraction of antistaling agent to treat these fruits,then put them in the above refrigerator. And used the equipment of vacuum and ozone to treat these fruits and put them in average temperature of 15 ℃,average humidity of 40% in house to keep fresh. Finding the best preservation method by measuring the weight loss rate,content of soluble solids and content of ascorbic acid(Vc)to determine the effects of various kinds of fresh-keeping. The results showed that the four kinds of preservation methods could significantly reduce the weight loss rate, delay of soluble solid and Vc content decrease rate. The preservatives with glycerol 3 g, starch 4 g, gelatin 1.5 g, agar 1 g, sodium metabisulfite 1.5 g mixed with distilled water to prepare a 500 mL was the best way of preservation. The vacuum and ozone preservation experiment of fruits of Kadsura coccinea, the vacuum preservation effect was better than ozone.

Effects of shading on contents of mogrosides and sugars in fruit flesh of Siraitia grosvenorii

2016/12/2 23:49:59
With the common Siraitia grosvenorii variety of “Yongqing 1” as experimental material, we measured the changes of concentrations of mogrosides monomers, soluble sugars, glucose, fructose, sucrose and starch in the fruit flesh, as well as the activities of sucrose metabolism enzymes, including acid invertase(AI), neutral invertase(NI), sucrose synthase(in synthesis direction, SSS; in cleavage direction SSC), sucrose phosphate synthase(SPS)and the net photosynthetic rates, during the late development stage of fruit under different shading treatments. The results indicated that in comparison with control, severe shading(70% shading)resulted in significant increase of mogroside V content: 60 d after shading(90 d after pollination), the content of mogroside V in the fruit flesh of 70% shading treatment was 5.79%, significantly higher than that of control group(4.83%, P<0.01). Meanwhile, the concentrations of glucose, fructose were also much higher than those of controls(P<0.05). On the contary, the changes of concentrations of total soluble carbohydrates and sucrose were far different from those of mogroside V, glucose and fructose: 60 d after shading, the levels of total soluble carbohydrates, sucrose in the fruit flesh of 70% shading treatment were significantly(P<0.05 ), extremely significantly(P<0.01)lower than those of controls, respectively. On the other hand, shading treatment led to a decrease of net photosynthetic rate(P_n): 40 d after shading, P_n of leaves under 70% shading treatment was significantly(P<0.05)lower than that of controls. The above results implied that severe shading treatment could result in the improvement of fruit quality of S. grosvenorii. In additon, under shading treatment, the decrease of photosynthetic rate could be the major reason for the decline of soluble sugar content, while changes of sucrose, fructose and glucose contents should be mainly resulted from the changes of activities of SPS, AI and SSC. In addition, under severe shading treatment, fall of temperature could be regarded as the probable cause for the increase of mogroside V content. The research provides the theoretical basis for the production techniques and the quality control of S. Grosvenorii fruit.

Effects of medicament transfusion following trunks on florescence of longan

2016/12/2 23:49:59
Effects of potassium chlorate and paclobutrazol on florescence of longan were studied in the form of medicament transfusion following trunks, which could increase the utilization of regulators of longan as a new type of infusion technology. The results were as follows: the technology of potassium chlorate transfusion following trunks could not only make the longan bloom earlier, but also increase the proportion of female flower and the rate of flower formation, earing and fruit setting. For the sensitivity to potassium chlorate, Shixia longan had high sensitivity, but poor stability; Chuliang longan had high stability, but poor sensitivity; Guixiang longan had middle sensitivity and stability. Treatment effects of Shixia was the best as a whole. The contents of the treatment were as follows: three times treated, 7.5 L(1 g·L^-1)potassium chlorate applied every time. The flower formation rate, earing rate, female and staminate flower proportion and fruit setting rate of this treatment were 53.67%, 59.38%, 16.01 and 19.58%. Paclobutrazol transfusion following trunks could not only control shoot growth effectively, but also improve flowering of longan. The best concentration of paclobutrazol was 160 mg·L^-1, which made the earing rate, the proportion of female flower, spica length, spica width and fruit setting rate of longan reach 96.53%, 10.36%, 21.95 cm, 12.96 cm and 38.37%. The technology of medicament transfusion following trunks applying potassium chlorate and paclobutrazol had positive effects on flowering and fruit setting of longan, which had reference function for achieving efficient, laborsaving, and environmental orchard management mode.

Establishment of regeneration of Actinidia arguta ‘Tianyuanhong'

2016/12/2 23:49:59
‘Tianyuanhong' was a new Actinidia arguta cultivar, with all red pericarp and pulp. In order to establish the rapid and efficient propagation system in vitro in ‘Tianyuanhong', we used stem with axillary bud, young leaf and leaf stem of ‘Tianyuanhong' as the explants, and used tissue culture methods to study explant types and the best plant growth regulator substance combination in this cultivar. The result showed that stem with axillary bud that sprout in early spring was the best explants, and MS+6-BA 0.5 mg·L^-1+IBA 1.0 mg·L^-1 was the suitable plant growth substances combination; leaf stem was the better experimental material than leaf to induce callus, MS+ZT 0.5 mg·L^-1 was the optimizing plant growth substances combination; the best plant growth substances combination in adventitious bud propagation was MS+ZT 1.0 mg·L^-1; and 6-BA could have the same effect in adventitious bud propagation, and could replace ZT for adventitious bud propagation in production, MS+6-BA 2.0 mg·L^-1+IBA 0.5 mg·L^-1was well; 1/2 MS+NAA 0.2 mg·L^-1was the best plant growth substances combination for rooting. After the seedlings rooted, transplanted the seedlings to the substrate, the seedlings could reach the 98% survival rate in the substrate with the ratio of perlite: peat soil: sand=1:1:1. Through this study, the in vitro regeneration system was established in all-red A. arguta ‘Tianyuanhong', which provides a good technical support for seedling micropropagation and factory nursery, at the same time, the established regeneration system provides the basis for the research of genetic transformation in all-red A.arguta.

Bioinformatics and the expression pattern analysis of soybean PM34 gene

2016/10/30 19:46:39
In this paper, the structure and tissue-specificity of the soybean seeds mature protein gene( PM34 )were studied, which could become the the foundations for further research the function of PM34 gene in soybean or the other plants. The sequence of soybean(Jidou2)PM34 gene was amplified by RT-PCR with the total RNA of soybean leaf as templet. The structure of soybean PM34 gene was predicted by bioinformatics. The expression pattern of soybean PM34 gene in soybean organs was detected by real-time quantitative PCR(qRT-PCR). The results showed that the molecular weight of the protein was 31.7 kDa, the isoelectric point was 6.60, and belonging to the hydrophilic protein. There was absent transmembrane domain and singal peptide. The secondary structure was comprosed of 12.97% alpha helix, 41.30% coil and 45.73% extended strand. The protein tertiary structure prediction showed that the homologous model template was 3 ijr. 1. A, which was a kind of short chain dehydrogenase/reductase and the homology was 54%. The evolutionary relationship demonstrated that soybean had relatively closer relationship with Vigna radiata and Medicago trun-catula than others. In addtion, the result showed that there was a little activity in roots, stems, leaves and flowers, but there was the highest activity in seeds, especially in mature seeds.

Cloning, sequence analysis and prokaryotic expression of LaMCT gene from Lepidium apetalum

2016/10/30 19:46:39
The active components of Lepidium apetalum, cardiac glycosides have been well studied both chemically and pharmacologically, but little is known about the cardiac glycosides biosynthesis pathway. Using cultivated seedlings as material, the transcriptome data of Lepidium apetalum, designing specific primers and using PCR method, an open reading frame(ORF)of key enzyme 2-C-methylerythritol-4-phosphate cytidylyltransferase(MCT)gene which involved in MEP pathway was isolated and named as LaMCT (GenBank accession no. KT832554). The sequence analysis and prokaryotic expression were also performed. Sequence analysis showed that LaMCT has an ORF of 912 bp, which encoded a protein of 304 amino acid residues. Subcelluar localization and conserved domain analysis indicated that LaMCT protein located in chloroplast had no signal peptide and transmembrane domain, but had a isoprenoid synthase conserved domain. Phylogenetic analysis revealed that LaMCT protein showed the highest homology, 94% similarity, with MCT protein from Arabidopsis thaliana. Through the construction of pET-32a-LaMCT vector, the recombinant LaMCT protein was successfully expressed in Escherichia coli BL21(DE3)cells. Finally, the recombinant LaMCT protein was purified through Ni²⁺ affinity chromatography. The LaMCT gene was cloned from L. apetalum, and the stable prokaryotic expression system of pET-32a-LaMCT was constructed. This study will provide some basic information for the further antibody preparation of LaMCT protein and would be helpful in functional research of LaMCT gene involved in cardiac glycosides biosynthesis pathway.

Effects of the infection with Xanthomonas campestris pv. phaseoli with the mutation in AhpC on chlorophyll fluorescence characteristics of bean leaves

2016/10/30 19:46:39
After the bean(Phaseolus vulgaris)leaves were infected with two different genotypes of Xanthomonas campestris pv. phaseoli, either wild-type(Xcp-W)or the mutant lacking Alkyl hydroperoxide reductase subunit C(Xcp-ahpC), the changes of the chlorophyll fluorescence parameters in both the infected site and the locations distant from the infected sites were studied. The results showed that in the infected site and the locations distant from the infected sites, the potential maximal photochemical efficiency(F_v/F_m)did not significantly change, while photosynthesis system Ⅱ(PSⅡ), actual photochemical efficiency [Y_(Ⅱ)], electronic transmission rate(ETR), and the coefficient of photochemical quenching(q_P)were significantly decreased. The decreases of PSⅡ, Y_(Ⅱ), ETR, and q_P in the leaves infected with Xcp-ahpC were more drastic than those in the leaves infected with Xcp-W. The quantum yield of non-regulated energy dissipation(Y_(NO))and quantum yield of regulated energy dissipation(Y_(NPQ))of the leaves were not significantly affected by the infection with Xcp-W. But,Y_(NO) and Y_(NPQ) of the leaves were significantly increased by the infection with Xcp-ahpC. These results showed that the infection with bacterial blight pathogen can cause a systemic inhibition of the photochemical efficiency of PSⅡ in bean leaves. And, this inhibition was more drastic when the leaves was infected by the pathogen lacking the Alkyl hydroperoxide reductase subunit C.

Effects of different antibiotics on callus differentiation and growth of Pogonatherum paniceum

2016/10/30 19:46:39
In the study, Pogonatherum paniceum calluses were selected as the experimental materials, and the effects of different kinds of antibiotics(kanamycin, hygromycin, ceftiofur sodium and ampicillin)and its concentrations to the growth and differentiations of P. paniceum calluses were compared and analysed in order to find out the suitable resistance selection agent and antibacterial agent in genetic transformation system of P. paniceum, respectively. The results were as follows:(1)P. paniceum callus had a strong sensitivity to kanamycin and the differentiation rate significantly decreased as the concentration of kanamycin increased(P=0.01). When the concentration of kanamycin was 10 mg·L^-1, the growth and differentiation of P. paniceum callus were significantly suppressed and differentiated a large number of whiten seedlings, but the differentiation rate was still at 36.56%. When the concentration of kanamycin was 15 mg·L^-1, differentiation rate of P. paniceum callus was at 11.94%, and only part of calluses were differentiated into green seedlings. However, when the concentration of kanamycin was 20 mg·L^-1, P. paniceum callus almost turn brown or dead, and the differentiation rate was only at 2.26%. Hence, 15 mg·L^-1 of kanamycin was fit for serveing as the resistance selection agent in genetic transformation system of P. paniceum.(2)P. paniceum callus had a less strong sensitivity to hygromycin than kanamycin. The effects of hygromycin on differentiation rate of P. paniceum callus was very small, but the toxicity was big, so hygromycin was not suitable as the resistance selection agent in genetic transformation system of P. paniceum.(3)300 mg·L^-1of ceftiofur sodium and ampicillin had a little effect on the growth and differentiation of P. paniceum callus, and all were able to inhibit the bacterials effectively. Morever, the inhibition of little higher level of ampicillin to the growth and differentiation of P. paniceum callus was less obvious. Therefore, 300 mg·L^-1of ceftiofur sodium and little higher level of ampicillin could be used as the antibacterial agent in genetic transformation system of P. paniceum as well. This study found out the suitable kinds and concentration of antibiotics served as the resistance selection agent and the antibacterial agent in Agrobacterium tumefaciens mediated genetic transformation system of P. paniceum, respectively, which provides the reference for studing genetic improvement and functional genes of P. paniceum.

Cloning and expression analysis of F3H gene in Caragana korshinskii

2017/7/2 22:58:41
Caragana korshinskii is a kind of shrub belonging to the leguminosae category and it has characteristic of tolerance to stressful conditions. Flavonoids are natural antioxidants that include the groups of notable pigments such as anthocyanins. The adversity stress can influence the biosynthesis of anthocyanins in plants. Flavanone 3-Hydroxylase(F3H)is a key enzyme needed for the biosynthesis of anthocyanins. In this study, we successfully isolated, cloned F3H gene from C. korshinskii. The full length cDNA of C. korshinskii F3H gene(designated as CkF3H)had an open reading frame(ORF)of 1 095 bp encoding 364 amino acids with a calculated molecular mass of 41.3 kDa as well as an isoelectric point of 5.9. Comparative and bioinformatical analyses revealed that the cloned CkF3H from C. korshinskii has high identity with F3H from other plant species. The deduced CkF3H protein showed similarities with other available plant F3H proteins. The promoter of CkF3H gene was isolated by chromosomal walking and 468 bp sequence was obtained by sequencing. PlantCARE analysis of this sequence showed that the peomoter contained some typical elements CAAT-box and TATA-box and kinds of Cis-acting elements involved in defence and stress responsiveness. RT-PCR analysis revealed that CkF3H was expressed in roots, stems, and leaves with almost no tissue specificity. The transcript level of CkF3H was increased in response to cold, high salt, drought and high temperature stress. The gene expression of CkF3H was influenced by photoperiod in response to cold stresses. The results suggested that CkF3H played an important role during cold, high salt, drought and high temperature stress in C. korshinskii.

Alleviation of Al toxicity by exogenous chlorogenic acid in black soybean

2017/7/2 22:58:41
We investigated the influences of exogenous chlorogenic acid(CGA)on the physiological parameters and Al stress-related gene expression of SB(Al-sensitive black soybean)root under Al stress. A major goal of this report was to explore the alleviating molecular mechanism of exogenous CGA on Al toxicity in SB. SB seedings were cultured in water solution and treated with 50 μmol·L^-1 Al or CGA plus 50 μmol·L^-1 Al. The optimal concentration of CGA alleviating Al toxicity was screened. And we study the effects of optimal exogenous CGA on root tip Al content, the activities of antioxidant enzyme, the expressions of 14-3-3 protein and H⁺-ATPase, and the activity of H⁺ pump under 50 μmol·L^-1 Al stress. Low concentrations of CGA alleviated inhibition of root elongation caused by Al toxicity and promoted the increase of lateral root numbers under Al stress. However, the CGA-alleviated effect was weaken with high concentration of CGA treatment. It was founded that the alleviation effect of 0.01 g·L^-1 CGA was the most significant. Exogenous 0.01 g·L^-1 CGA decreased the Al in root tips and MDA contents. And the citric acid content in root exudates of SB under Al stress was significantly increased due to the addition of exogenous 0.01 g·L^-1 CGA. Expression analysis showed that exogenous 0.01 g·L^-1 CGA enhanced the expression of three 14-3-3 isoforms(14-3-3b, 14-3-3m, and 14-3-3k)and increased the expression of GHA2 (plasma membrane H⁺-ATPase)gene, but inhibited the gene expression of MATE in SB under Al stress. The results of immunoprecipitation(IP)showed that the phosphorylation of the PM H⁺-ATPase was up regulated by the exogenous 0.01 g·L^-1 CGA, which could bind with 14-3-3 protein under Al stress. Meanwhile, plasma membrane H⁺-ATPase and H⁺ pump activities were both enhanced by exogenous 0.01 g·L^-1 CGA under Al stress. It was suggested that exogenous CGA may enhance the SB tolerance to Al stress by increasing the lateral roots number, compensating inhibition of MATE expression and interaction between them to increase citric acid exudation.

Effects of abscisic acid on PLD activity in mitochondrial membrane under low temperature

HE Wen-Ping, ZHANG Xu-Qiang, YANG Peng-Jun, QIU Yi-Zhi, WANG Xin-Xia, YANG Ning — 2017/7/2 22:58:41
Choraspora bungeana is a kind of rare alpine periglacial plant. The environment that C. bungeana lives in has many stress factors such as low temperature, strong ultraviolet. PLD plays an important role in degradation of membrane phospholipids and signal transduction of phospholipids. But the activity of PLD is affected by many factors usually. In this study, the plantlets of C. bungeana was used as experimental materials to study the effects of abscisic acid(ABA)on the mitochondrial membrane bound-PLD activity in the leaves of plantlets of C. bungeana under low temperature(4, 0 and -4 ℃). The results indicated that when plantlets of C. bungeana was treated by 10, 50 and 100 μmol·L^-1 ABA, the mitochondrial membrane bound-PLD activity was higher than that in the group without ABA. Among these concentrations of ABA, 50 μmol·L^-1 ABA was more effective to promote the mitochondrial membrane bound-PLD activity compared with ABA of other concentrations. Sodium tungstate was an inhibitor of endogenous ABA biosynthesis and EGTA was a chelating agent of Ca²⁺, both of them had an important role in researching the signal transduction of ABA and Ca²⁺. When the plantlets of C. bungeana was treated by 0.3 mmol·L^-1 Sodium tungstate, the mitochondrial membrane bound-PLD activity in the leaves of plantlets of C. bungeana was lower than that in the control without sodium tungstate. When the plantlets of C. bungeana was treated by 50 μmol·L^-1 ABA+ 5 mmol·L^-1 EGTA, the mitochondrial membrane bound-PLD activity in the leaves of plantlets of C. bungeana was lower than that in the group without EGTA. When the plantlets of C. bungeana was treated by 0.3 mmol·L^-1 sodium tungstate + 10 mmol·L^-1CaCl₂, the mitochondrial membrane bound-PLD activity in the leaves of plantlets of C. bungeana was higher than that in the group without CaCl₂. From above results, ABA could affect on the mitochondrial membrane bound-PLD activity in the leaves of plantlets of C. bungeana by Ca²⁺ under low temperature.

Regeneration of leaves of noni

2017/7/2 22:58:42
Using noni(Morinda citrifolia)leaves as explant for culture in vitro on 3% MS medium with different types and concentrations of plant hormones, two kinds of in vitro regeneration modes were built. Mode Ⅰ: the callus was induced first, and then the adventitious roots and buds were induced; Mode Ⅱ: the adventitious roots were induced, and then the buds were induced directly. The results showed that the optimal medium of generating callus by noni leaves in Mode I was MS + 0.1 mg·L^-1 6-BA + 2.0 mg·L^-1 2,4-D; the optimal medium that induced leaf callus to generate adventitious roots and buds was MS + 1.0 mg·L^-1 6-BA+ 0.4 mg·L^-1 NAA or MS + 2.0 mg·L^-1 6-BA + 0.4 mg·L^-1 NAA, Thereinto, the solution of MS + 1.0 mg·L^-1 6-BA + 0.4mg·L^-1 NAA made the rooting time earler, about 10 d, and its root system was more developed. Instead, the solution of MS + 2.0 mg·L^-1 6-BA + 0.4 mg·L^-1 NAA made it 15 d. The optimal medium that induced leaves to generate roots and buds in mode Ⅱ was MS + 1.0 mg·L^-1 6-BA + 0.4 mg·L^-1 NAA。Cutting and transplanting the seedlings regenerated in vitro from model Ⅰ and Model Ⅱ into MS + 0.2 mg·L^-1 NAA medium to induce rooting. Getting the differentiation of adventitious root about 15 d and complete plant 45 d. This study provides useful references for the breeding and the application of genetic transformation technique in noni.

Effects of clonal integration on the nitrogen availability of rhizosphere soil in Phyllostachys nigra suffering from heterogeneous light

2017/7/2 22:58:42
To study the effects of clonal integration on SOC, TN, DOC, DON, NH₄⁺-N, NO₃^--N and microbial community composition, a pot experiment was conducted by rhizome clonal plant Phyllostachys nigra fragments consisting of two successive ramets subjected to heterogeneous light. Mother ramets suffering from full sun versus offspring ones suffering from 80% shade, meanwhile, the rhizome was kept connected or severed. The results showed that SOC, TN, DOC and NH₄⁺-N concentrations of shaded P. nigra offspring ramets showed a significant difference between connected and severed treatments, which means clonal integration improves soil nitrogen utilization effectiveness for connected shaded offspring ramets; Principal component analysis of PLFAs concentration of rhizosphere soil microbial for P. nigra showed a significant difference between the two treatments of shading connected and severed conditions. The results indicated that P. nigra could reduce the percent of bacteria in soil which nitrogen utilization effectiveness was low, and could increase the percent of actinomycetes and fungi by clonal integration, so as to improve the soil nitrogen utilization efficiency of whole P. nigra, which endued them with the adaptive advantages to heterogeneous habitats.

Composition of mycorrhizal fungi and symbiotic relationship of Cypripedium yunnanense

AN Man-Yun — 2017/7/2 22:58:42
Cypripedium plants are endangered by over-harvesting due to their high ornamental and medicinal value. It is necessary to obtain the culturable mycorrhizal fungi in the application of mycorrhizal technology for the protection and artificial cultivation of C. yunnanense. The composition of mycorrhizal fungi and their symbiotic relationship of C. yunnanense were studied by culture-dependent and inoculate methods. The results showed that one hundred and twenty-six independent fungal isolates were obtained from three hundred root tissues of C. yunnanense. The isolates were identified to three genera: Tulasnella (seventy-three), Corticium (thirty-six)and Ceratobasidium (seventeen). Among them,Tulasnella(57.94%)were dominant genus. The mycorrhizal fungi could significantly shorten the seed germination process and promoted the seeding growth. The strain CY-18 was found to be a growth-promoting fungus. Moreover, above results simultaneously could be used as a valuable candidate sources for the protection and sustainable utilization of C. plants resources by mycorrhizal technology.

Cloning and expression analysis of tryptophan aminotransferase of Arabidopsis 1-Related 2

2017/4/7 11:47:51
Tryptophan aminotransferase gene family is a key gene that directly involved in auxin biosynthesis pathway in plants. The full-length cDNA sequence and DNA sequence of Siraitia grosvenorii tryptophan aminotransferase of Arabidopsis 1-Related 2(SgTAR2)gene was amplified by using transcriptome sequencing data and the technology of rapid amplification of cDNA ends(RACE)method. Subsquently, analysis of the gene structure of SgTAR2 by bioinformatics, and the expression levels of SgTAR2 in different tissues of S. grosvenorii were analysed by real-time quantitative method. The results showed that cloned full length of SgTAR2 cDNA sequence was 2 078 bp, containing a 1 332 bp open reading frame(ORF)which encoded 443 amino acids, and GenBank accession number was KU949381. The coding protein had two alliinase domains and multiple 5'-PLP binding sites, which might take part in catalytic amino acid transfer, chemical defense, auxin biosynthesis and other biological processes. The full-length DNA sequence was 4 103 bp with four introns and five extrons, and these introns contained multiple 5UTR Py-rich stretch, hormonal and environmental stress responses related cis-acting elements, which might play a role in auxin biosynthesis, stress response, morphological development and other biological processes. The result of Real Time PCR exhibited that SgTAR2 was the highest abundance in pistils and 15 d fruit, and suggested that auxin synthesis was exuberant in initial development of S. grosvenorii fruit. Therefore, SgTAR2 plays a significant role in the growth and development of S. grosvenorii by auxin mediated, especially in young fruit and flower initial development and organ morphogenesis.

Gene cloning and molecular characteristics of squalene synthase from Cucumis melo

2017/4/7 11:47:51
Cucurbitacins(Cus)are tetracyclic triterpenoid compounds which exist mainly in cucurbitaceous plants and have various pharmaceuticals-like actions. The medicinal raw materials of Cus are currently extracted mainly from the pedicellus melo. In this study, the gene of squalene synthase(SQS), the key enzyme for Cus biosynthesis, from Cucumis melo was cloned, and the amino acid sequence of the enzyme derived from its nucleotide sequence was analyzed with bioinformatics methods. DNA sequencing and BLAST RefSeqGene analysis indicated that the cloned fragment of the C. melo SQS gene contained a complete open reading frame(ORF)of the gene. ORF Finder analysis showed that the SQS of C. melo was constituted of 417 amino acid residues with an isoelectric point of 7.56. Analysis of the deduced amino acid sequence suggested that the main type of secondary structure of the enzyme was α helix. Domain prediction study indicated that the SQS belonged to the isoprenoid biosynthase superfamily possessing the banding sites of farnesyl diphosphate and magnesian ion. Prediction of the tertiary structure suggested that the SQS was a monomeric enzyme with a cave-like activity center formed by α helixes. Protein phosphorylation analysis indicated that the phosphorylation site S⁴⁸ located in the activity center-related motif⁴⁷ VSRSF⁵² and S¹⁹⁶ was a positive selection site, suggesting that both S⁴⁸ and S¹⁹⁶ were critical sites for the regulation of SQS activity. The phylogenetic classification based on the phylogenetic tress constructed with the ORF sequence of the SQS gene showed that the result was in accordance with that of morphologic classification. Therefore, this study provides new clues and reference for research of the regulation of Cus biosynthesis.

Cloning and expression analysis on the ScALDH gene of Saccharum spontaneum

2017/4/7 11:47:51
We cloned a drought resistant ALDH gene with RT-PCR gene from Saccharum spontaneum, detected the gene expression level under different drought conditions, and analyzed the nucleotide and amino acid sequences with Blastx, Conserved Domain Search, ORF finder and MEGA. We compared the amino acids among different species, and tried to find reading frames in the sequence, to search conserved Domain and to make evolutionary tree. We analyzed the gene expression level of the cloned gene in drought stress with real time-PCR. The total length of the ALDH gene fragment cloned was 1 996 bp, including 1 524 bp coding sequence, which was encoded 508 amino acids. The cloned sequence had a very high homology with other species' amino acid sequences and had the conserved sequences of ALDH family, a complete open reading frame as well. Phylogenetic tree analysis showed that protein had the closest relationship with corn. The real time-PCR data showed that the expression of the gene was expressed in a continuous accumulation mode. On the whole, the gene responds to drought stress. The cloned gene belong to the ALDH protein family, which has its typical function domain. The expression pattern analysis showed that the gene was involved in the process of drought stress. The results of this study aimed at developing the excellent resources of wild species, as well as to explore the drought resistance of these resources. The study provides the basis for the selection of the parents of the fine and drought resistance in the future.

Separation and purification of plasma membrane from bark of Hevea brasiliensis by aqueous two-phase partition

2017/4/7 11:47:51
The plasma membrane H⁺-ATPase of rubber tree bark involved in various important physiological processes, such as ion transportation, stress responses, production and release of natural rubber latex, the preparation of a high purity and activity plasma membrane is the first and essential step to understand the functions of H⁺-ATPase in some in situ situations. In the present study, the separation of crude microsomes was isolated by differential and gradient centrifugations, the aqueous two-phase partition system was employed to separate the organelles membrane constitutions and receive highly pure plasma membrane, and the effect on the extraction efficiency of polymer and KCl concentration in partition was studied as considered the characteristic of bark of rubber tree. The total concentration of plasma membrane proteins was measured by Bradford method. The purity of different membrane was determined by the sensitivity of representative enzyme activity measured by Molybdenum blue method. H⁺-ATPase of plasma membrane was specifically inhibited by NaVO₃ under pH 6.5, due to different types of H⁺-ATPase in different organelle membranes, while H⁺-ATPase of tonoplast or plastid membrane was suppressed by KNO₃ or NaN₃ under pH 8.5, respectively. Our results indicated that the combination of 6.4%(W/W)polymer and 5 mmol·L^-1 KCl applied in two-phase partition system achieved the relatively high purity and yield ratio of plasma membrane whereas high polymer concentration of or KCl could increase the purity but with poor productivity. In order to further verify that morphological diversity between purified plasma membrane and crude microsomes, transmission electron microscopy(TEM)was performed. Paraffin embedded thin sections was made and stained separately with lead uranium and phosphotungstic acid. The morphological analysis showed that in crude microsomes there was little plasma membrane and full of cross-contaminant membrane substitutions, while after purified, the constitution of plasma membrane was relatively pure with less contaminant. In summary, highly purified plasma membrane was yielded by the aqueous two-phase partition system with 6.4%(W/W)polymer and 5 mmol·L^-1KCl and perform a comparatively singular morphology, which could be further used to reveal the function of H⁺-ATPase of bark of rubber tree in production and release of rubber and other physiological processes.

Correlation between fruit form and seed setting rate of Bletilla striata

2017/4/7 11:47:51
Bletilla striata is a perennial herb belonging to the family Orchidaceae, has been used as Chinese folk medicine. Seed quality is key to get large number of B. striata seedlings, and the seed setting rate is a very important standard to evaluate the fruit quality. It is a quick method to rough evaluate seed setting rate through the fruit form. In this study, the correlation between fruit form(fruit weight, length and diameter)and seed setting rates among 26 infructescences(87 fruits)of B. striatawere analysed, the seed embryo sizes of ten different sizes of B. striata fruits were measured, and the weights of 400 seeds were calculated and the weights of single seed were obtained. The results showed that correlation between fruit weight and seed setting rate were the most close, the coefficient of correlation was 0.786, reaching to moderate degree; the second close was fruit diameter, the coefficient of correlation was 0.634; and the last was fruit length, the coefficient of correlation was 0.553; the average setting rate of 87 fruits was 50.43%. The correlation between fruit form and seed embryo size was at a low degree, developmental state of seed embryo could not be known through the fruit form; the average diameter of embryo was 177.9 μm, the biggest was 241.9 μm, the smallest was just 64.5 μm; and the average length of embryo was 359.0 μm, the biggest was 528.6 μm, the smallest was just 114.3 μm, the results showed that the mature seed embryo size of B. striata mainly focus on 150-200 μm in diameter, the ratio was 62.34% in this range, and 300-400 μm in length, the ratio was 68% in this range. The average weight of single seed was 3.90 μg, the seeds in the biggest fruit of the 87 was 39 488, and the average number was 10 436, according to calculation, the average number of have embryo seeds in a single fruit was 5 263, it could gain a large number of B. striata seedlings through the sterile culture. In conclusion, fruit weight and full degrees(diameter)can be used to evaluate fruit quality(seed setting rate)of B. striata.

Effects of cultivation patterns and compound fertilizer types on agronomic traits and dry matter accumulation of Longping 206

2017/4/7 11:47:51
We studied the effects of four cultivation patterns and two compound fertilizer types on the root morphology index at seedling stage, the volume and the dry weight of root at elongation stage, the physiological property of root, leaf and stem, the agronomic traits and dry matter accumulation, grain filling and N, P and K accumulation, photosynthetic characteristics and yield components. This experiment was done at the science and technology garden in Anhui Science and Technology University and designed by two-factor split block design. The main plot was row-spacing allocation and double plants a hole. The four seeding styles were equal row spacing(A₁, 60 cm), equal row spacing and double plants a hole(A₂, 60 cm), wide and narrow row spacing(A₃, 70 cm+50 cm), wide and narrow row spacing(A₄, 90 cm+30 cm), respectively. The vice plot was base fertilizer type and used Stanley compound fertilizer and Difuyuan special compound fertilizer for corn. The experiment had eight treatment groups and four repetitions for each group. The seeding density was 67 500 cobs·hm^-1, the width of each plot was 3 m, and the length was 6.67 m. The underground fertilization amount was 750 kg·hm^-1. The plant height and growth rate of Longping 206 had not significant difference between equal row spacing and wide and narrow row spacing, but had significant difference during the pattern of equal row spacing with double plants a hole. The ear height had not significant difference between A₁ and A₃, while had significant difference between A₄ and A₂. The stem diameter had not significant difference between each treatment. The effects of cultivation patterns on the angle between first leaf of above ear and first leaf of under ear had significant difference, and on the first leaf orientation value of above ear also had significant difference, but on the first leaf orientation value of under ear had not significant difference. The effects of cultivation patterns on the stem intensity of ear knob and upper ear knob and on the dry matter accumulation at different stages had significant difference. Longping 206 in wide narrow row spacing(90+30)cm, Difuyuan special compound fertilizer, the indexes of agronomic characters of long reached the best state, the effects of cultivation pattern of dry matter accumulation of significant difference of Longping 206 on different periods, at jointing stage, booting, flowering stage, milk stage and mature period, the amount of dry matter accumulation of Longping 206 were the biggest in cultivation mode row(70+50)cm. It illustrated that different cultivation patterns and compound fertilizer types made important effects on the growth of Longping 206 and provided some information for cultivation management and fertilization of Longping 206.

Cloning and bioinformatics analysis of RCA2 gene in Dendrobium officinale

JIANG SuHua1, ZHANG Yan1, WANG MoFei1, WANG JieQiong2, CUI Bo1* — 2017/2/9 11:03:18
Ribulose1,5bisphosphosphate carboxylase/oxygenase was a key enzyme of photosynthetic carbon assimilation, it had direct adjustive effects Rubisco activity and photosynthetic rate of in Dendrobium officinale, and laid a good foundation for changing plant photosynthetic rate by studying the gene. RCA gene was cloned from leaf of annual D. officinale using RTPCR and RACE, and was analyzed by bioinformatics. The results showed that the full length cDNA of RCA was 1 730 bp length, the cloned RCA gene was named RCA2(GenBank accession No. KT 205842). It was contained a 5′untranslated region (5′UTR) (81 bp), 3′UTR (326 bp)，and an opening reading frame (ORF) (1 323 bp), which could be translated into a 410aminoacid putative peptides with a molecular weight of 48.53 kDa and a theoretical pI of 6.19, with a domain of Ploop_NTPase superfamily. Multiple sequence alignment results showed that the homology of nucleotide sequence between the RCA2 and the reported Phalaenopsis genes was 87%. RCA2 protein was belong to hydrophilic protein，subcellular localization was in the chloroplast. The RCA2 protein secondary structure components showed that ahelix(h), extended strand(e), and random coil (c) accounted for 30.68%, 25.45% and 43.86%. RCA2 encoding protein function prediction showed that RCA2 played a very important role in intermediary metabolism. This study of the key enzymephotosynthesis Rubisco in Dendrobium officinale provides the information for the research of the photosynthetic characteristics, and the basis for improving the efficiency of greenhouse cultivation.

Construction of potato tetravalent antivirus plant expression vector and its tobacco transformation

2017/2/9 11:03:18
In order to simultaneously foster antifourvirus potato varieties(PVX, PVS, PVY and PLRV), four different viral coat protein (CP) gene PVXCP (670 bp), PVSCP(800 bp), PVYCP(700 bp) and PLRVCP(600 bp) were obtained via RTPCR and sequenced to confirm respectively. And the comparison of the sequences that we obtained and the already reported sequences from NCBI database showed that all four viral CP genessequences were more than 90% homologous; then around 300 bp size conservative gene fragments PVXrh, PVSrh, PVYrh, PLRVrh were selected from their respective viral CP genesand four specific bands which were consistent with the fragment size were obtained via PCR amplification. The fusion sequence XSYVrh which is around 1 200 bp long was created with PVXrh, PVSrh, PVYrh and PLRVrh gene fragments via OverlapPCR technique. With the help of DNA recombination technique, we integrated XSYVrh sequence into pGMT vector and created cloning vector pGMTXSYVrh. The cloning vector pGMTXSYVrh and the plant expression vector pART27 were treated with incision enzyme Spe Ⅰ and Sac Ⅰ; then under the effect of T4 DNA ligase, XSYVrh sequence was integrated with pART27 expression vector; at last pART27XSYVrh plant expression vector was successfully constructed. The pART27XSYVrh plant expression vector was introduced into Agrobacterium tumefaciens LBA4404, and subsequently introduced into tobacco (T12) assisted by the agrobacteriummediated transformation. PCR results showed that there were 40 transgenic plants with targeted gene integrated into their genomes.

Effects of three plantgrowth regulators on the growth of pseudobulbs in Bletilla striata seedlings

2017/2/9 11:03:18
Effects of Brassinolide (BR), Naphthaleneacetic acid(NAA) and Methyl Jasmonate(MeJA) on the growth of pseudobulbs in Bletilla striata seedlings were studied by measuring biomass, amino acid contents, protein contents and polysaccharide contents with annual seedlings, and economic efficiency were further analyzed. The results showed that the pseudobulbs weight of B. striata treated by 1.6×104 mmol·L1 BR was 3.39 g·FW, which was the highest among all treatments, and it was 1.19 times and 1.25 times of water treatment (CK1) and biogas fertilizers treatment (CK2), respectively. At the same time, the new bud number was 2.17 for B. striata treated by 1.6×104 mmol·L1 BR, which was also higher than other treatments. The results indicated the production efficiency will be improved at least 19% treated by 1.6 ×104 mmol·L1 BR for B. striata. In comparison to CK1 and CK2, amino acid contents, protein contents and polysaccharide contents of the pseudobulbs of B. striata treated by 1.6 ×104 mmol·L1 BR treatment, 0.5 mmol·L1 NAA treatment and 0.25 mmol·L1 MeJA treatment were further measured, respectively. It was 5.21% and 8.58% for the amino acid contents of CK1 and CK2, respectively. And it were 7.26%, 7.53% and 5.69% for that of 1.6 ×104 mmol·L1 BR, 0.5 mmol·L1 NAA and 0.25 mmol·L1 MeJA, respectively. The protein contents were the same to that of amino acid contents from high to low for the pseudobulbs of B. striata, that was CK2＞1.6×104 mmol·L1 BR＞0.5 mmol·L1 NAA＞0.25 mmol·L1 MEJA＞CK1, the contents were 11.6%, 11.0%, 10.5%, 9.14% and 7.72%, respectively. However, the polysaccharide contents treated by 1.6 ×104 mmol·L1 BR, 0.5 mmol·L1 NAA and 0.25 mmol·L1 MeJA were 24.2%, 26.5% and 26.5%, respectively, the three treatments were all higher than that of CK1(19.3%) and CK2(21.8%). In conclusion, not only the yields of pseudobulbs were promoted but the quality characteristics were improved if B. striata were treated by 1.6 ×104 mmol·L1 BR. This plays an important role in the large scale planting of B. striata in the future.

Callus inducing on different phytohormone media and Agrobacteriummediated genetic transformation system optimizing of Salvia miltiorrhiza

ZHANG LinSu1*, SHAO FenJuan2, QIN LiJun3 — 2017/2/9 11:03:18
With the decoding of its genome, Salvia miltiorrhiza, belonging to labiatae (Lamiaceae) sage (Salvia Linn), is becoming an important model medicinal plant and is being widely studied. The earliest record of S. miltiorrhiza can be found in more than two thousand years ago in the Sheng Nong’s Herbal Classic in which S. miltiorrhiza was listed as one of the top grade drugs. S. miltiorrhiza has a long history in treatment of many cardiovascular and cerebrovascular diseases. The active components of S. miltiorrhiza are divided into two categories: watersoluble phenolic acids and lipidsoluble tanshinones. As an important medicinal mode plant with long history, the biosynthetic pathways of S. miltiorrhiza active contents tanshinones and phenolic acids attract growing research interests. An appropriate tissue culture condition and a simple, stable and efficient genetic transformation system are very important in the research of the plant S. miltiorrhiza. Based on former studies, we investigated culture conditions to induce different S. miltiorrhiza calluses using different phytohormones on Murashige Skoog medium. And through a four factorthree level orthogonal design, we investigated the effects of submerging time, coculture time, screening pressure and concentrate of acetosyringone. The results of callus induction were that: MS+0.5 mg·L1 6BA+0.5 mg·L1 2,4D induce callus faster than other explants, while MS+0.1 mg·L1 NAA+0.5 mg·L1 6BA was suitable in inducing stems; 1 mg·L1 transzeatin could induce brawn callus maybe contain more tanshinone, 1.0 mg·L1 2,4D could induce more roots on callus. And the confirmed agrobacteriummediated genetic transformation parameters for agrobacterium tumefaciens GV3101 harboring nptII gene plasmid were: submerging 5 min, coculture 1 d and screening at 30 mg·L1 kanamycin, the positive ratio of transgenic plant by PCR verification was 60%. When submerging 5 min, coculture 1 d and screening at 10 mg·L1 streptomycin the transgenic plant positive rate was 70%. The later transgenic system seemed more stable and easier to be repeated in this trial, maybe due to the less toxic of streptomycin which indicated that streptomycin maybe another candidates for transgenic screening in S. miltiorrhiza. The determination of different callus inducing condition and the optimizing of genetic transformation system provide useful appliancation in further researches of S. miltiorrhiza.

Biological characteristics of five rare and endangered Camellia seeds in Guizhou Province

LV XiaoMei1, LIU HaiYan2, HUANG LiHua3, YANG NaiKun4, ZOU TianCai3* — 2017/2/9 11:03:18
Camellia longistyla, C. delicata, C. kweichouensis, C. rhytidophylla, C. luteoflora are rare and endangered species with important economic value. They are locally distributed in evergreen broadleaved forest and evergreen and deciduous broad leaved mixed forest in Guizhou Plateau (ⅢD10 d). In order to explore the seed reproduction and endangered reason of these species， the biological characteristics were studied in this paper. The results were as follows: (1) About thousandgrain weight and germination rate, the maximum was C. longistyla 3 289.70 g and 81.67%, the minimum was C. kweichouensis 786.33 g and 46.00%. (2) The maximum water content was C. luteoflora 48.85%, the minimum one was C. longistyla 39.52%. (3) The maximum viability was C. longistyla 98.33 %, the minimum one was C. rhytidophylla 63.33%. (4) The results indicated that thousand grain weight was significantly positively correlated with viability, germination rate, germination energy and germination index (R≥0.772， P＜0.01). The viability and germination rate were positively correlated with germination energy significantly (R≥0.738， P＜0.01) and positively correlated with germination index (R=0.532， P＜0.05). The seed coat was hard and not easy to absorb water, and it was not easy for the population reproduction because of the loss of feeding and disease and insect erosion, which was an important reason for the endangered species. The experimental results provides the reference for the protection and utilization of these plant resources.

Ariation in traits of cone and seed of Keteleeria fortunei var. cyclolepis and its relationship with environmental factors

2017/2/9 11:03:18
This study focused on the research of Keteleeria fortunei var. cyclolepis in transitional regions from lowlying basin of Guangxi to the YunnanGuizhou plateau. The purpose was to discuss the diversity in phenotypic traits of cone and seed among populations of K. fortunei var. cyclolepis and its relationship with geographical environmental factors. We made a comparative study on sixteen phenotypic traits of cone and seed in nine natural populations of K. fortunei var. cyclolepis from four provinces. To analyze the experiment data, the main mathematical statistical methods were used including variance analysis, multiple comparison, correlation analysis, and stepwise regression analysis. The results showed that there were abundant variations in phenotypic traits of cone and seed among populations of K. fortunei var. cyclolepis. And there were significant or extremely significant differences in sixteen phenotypic traits of cone and seed among populations. The variation coefficient of natural population in Wangmo was the highest (2.54%) among nine natural populations. The variation coefficient of seed width was the highest (9.04%) among sixteen phenotypic traits. The variation of seed traits was higher than that of cone among populations. It meant that the cone has higher stability. There were significant or extremely significant correlations among part of phenotypic traits of cone and seed. The following phenotypic traits including seed width, seed thickness, seed aspect ratio, seed scale aspect ratio and bract aspect ratio were more important traits. To some extent, they could reflect the quality of cone and seed. Seed traits were more largely influenced by geographical environmental factors compared to cone traits. The seeds in west area were near oval and more satiate. And the seed wings were longer and wider of the seeds in the west than the seeds in other areas. There were significant correlations between elevation and majority traits of cone and seed. Analyzing the outcomes of these experiments, we can draw the conclusion that elevation as a major variation source is the main factor affecting variation in phenotypic traits of cone and seed of K. fortunei var. cyclolepis among geographical environmental factors. These results offer information in protecting and taking advantage of germplasm resources of K. fortunei var. cyclolepis.

Effects of nitrogen addition on seedling growth and biomass allocation of three rare tree species in the south of China

2017/2/9 11:03:18
In order to investigate the shortterm response of rare tree species to nitrogen addition, the seedling growth and biomass allocation of Semiliquidambar cathayensi, Tsoongiodendron odorum, Symplocos tetagona under five soil nitrogen levels(0, 0.1, 0.2, 0.4 and 0.6g · kg1soil) were studied for more than four months. The results showed that the seedlings of the three tree species responded differently to the different nitrogen content levels. The plant height, basal diameter, crown width, total biomass and the biomass of each organs increased significantly in the seedlings of Tsoongiodendron odorum. Low and medium nitrogen promoted and high nitrogen inhibited the growth of Semiliquidambar cathayensis seedlings. The morphology and growth parameters of Symplocos tetagona seedlings were not significantly different among low nitrogen contents, but were inhibited under medium amount of nitrogen. The biomass allocation of three species seedlings varied significantly among different nitrogen treatments. With the increase of nitrogen levels, the root mass ratio and the ratio of root / shoot of Tsoongiodendron odorum seedlings decreased significantly, and those parameters of Semiliquidambar cathayensis seedlings increased significantly with the increase of nitrogen levels except for the highest nitrogen treatment. The root biomass ratio and the ratio of root / shoot of Symplocos tetagona seedlings increased significantly with the nitrogen content, which may relate to the growth of stem and leaf that were inhibited under medium and high nitrogen levels. Overall, Tsoongiodendron odorum seedlings were more resistant to high nitrogen loads, Semiliquidambar cathayensis seedlings came to the second, and Symplocos tetagona seedlings were not resistant to high nitrogen loads. At the end of the growing season, the height, basal diameter and total biomass relative growth rate of Semiliquidambar cathayensis seedlings were significantly higher than those of the other two plant species.

Dynamic changes of endogenous hormones in fruit branch leaves of Camellia nitidissima with different flower numbers during blossom and fruit periods

2017/12/28 18:19:56
In order to investigate the physical problem of flowering and fructification rate of endangered plant Camellia nitidissima, and provide the theory basis for species conservation, the contents and ratio of GA₃, IAA, ZR and ABA in fruit branch leaves of C. nitidissima of more-flower, less-flower and non-flower were measured by ELISA(Enzyme-linked immunosorbent assay)method during blossom and fruit periods. In early-bloom stage, the content of IAA in leaves rised firstly and then fell, and its content of more-flower-plant was lower than that of less-flower-plant leaves; the content of ZR in flower-plant was higher than that of non-flower-plant; the content of GA₃ appeared an upward trend, and its content of flower-plant was higher than non-flower-plant; the contents of ABA decreased firstly and then increased, and its content of more-flower-plant was lower than non-flower-plant. In full-bloom stage, the contents of IAA, GA₃ and ABA declined overall; the content of ZR rised firstly and then fell; the contents of IAA, ZR, GA₃ in more-flower plant were higher than those in less-flower-plant or non-flower-plant, while the content of ABA was less than that of other plants. During the bloom period, the ratio of IAA/ZR, IAA/ABA, ZR/ABA and GA₃/ABA of flower-plant were higher than that of non-flower-plant, but the ratio of(IAA+GA₃)/ZR was just in contrast, which also indicate the development of flower and fruit was related to not only single plant hormone but also the balance of plant hormones. During autumn shoot period with active vegetative growth, the ratio of IAA/ZR of non-flower-plant was larger than that of flower-plant, while the ratio of IAA/ZR of non-flower-plant was smaller in blossom and fruit period with vigorous reproductive growth, but the ratio of(IAA+GA₃)/ZR was just in contrast. The results showed that high level of IAA, ABA, ZR and low level of GA₃ in bud stage was benefitial for blossom, high content of IAA, ZR and low content of GA₃, ABA in post-bloom stage was conducive to reducing blossom and fruit dropping, increased fruit-bearing rate and promoting fruit rapid growth, high ABA content before harvesting was favorable toward fruit ripening. Therefore, it can provide theoretical reference for using plant growth regulator to reduce the fallen petal and fruit abscission in production of C. nitidissima.

Catalytic functional identification of cassava hexokinase MeHXK5

2017/12/28 18:19:56
Hexokinase is a key enzyme in the process of catalyzing the phosphorylation of hexose in plants. It is involved in the process of plant growth and development. We cloned hexokinase gene MeHXK5, which is mainly expressed in cassava flowers. In this study, we used the yeast mutant strain YSH7.4-3C to perform yeast functional complementation experiment to determine whether MeHXK5 had the function of catalyzing the phosphorylation of hexose. It was found that YSH7.4-3C yeast with pDR195-MeHXK5 vector could be grown in the medium with glucose or fructose as the sole carbon source, indicating that MeHXK5 could catalyze the phosphorylation of hexose to provide carbohydrate and energy for yeast growth. The results provide information for further study of the physiological functions of MeHXK5 in the flowering process of cassava.

Seed dormancy mechanism and its ecological significance of endangered species Dracaena cambodiana

2017/12/28 18:19:56
In order to understand the dormancy mechanism of Dracaena cambodiana seed, the dormancy types and methods for seed dormancy breaking were studied systematically, and the ecological significance for dormancy mechanism of D. cambodiana seeds were also investigated. The results showed that the hydroscopicity of D. cambodiana spermoderm was good with(42.60 ± 0.08)% seed imbibition rates after 40 h; excised embryos began to geminate after 2 d with a germination rate of 100%, suggesting a mature embryo for D. cambodiana; and the germination of seeds without micropylar structure was the same to excised embryos, which showed the main reasons for failure of germination was the mechanical resistances of the hard micropylar structure; there were inhibitory substances in the seeds since when cabbage seeds were treated with the water crude extracts of D. cambodiana seeds for about 48 h, the seed germination and seedling growth of cabbage was inhibited obviously. Both rapid dehydration and storage under low temperature can break effectively the dormancy of D. cambodiana seeds, to increase the germination rate to 100% and germination uniformity. The analysis suggest that the growth potentials of the embryo maybe limited by seed-inhibitory substance are not able to break the mechanical resistances of the hard micropylar structure, and this is the dormancy mechanism of D. cambodiana seeds, which belong to physiological dormancy; the seed germination and seedling growth of D. cambodiana with the seed dormancy can adapt to the seasonal change of habitat conditions, which ensure the exist and regeneration of D. cambodiana population.

Different adaptations to salt stress in different ploidies of wheat

2017/12/28 18:19:56
In order to investigate different adaptions to salt stress in different ploidies of wheat, K⁺ and Na⁺content, K⁺/Na⁺ratio, the activity of SOD, CAT, POD and the contents of MDA, soluble protein, and soluble sugar were analyzed in synthetic hexaploid(AABBDD)wheat and its tetraploid(AABB)wheat(Triticum turgidum)and diploid(DD)(Aegilops tauschii)parents after treated with 0, 50, 100, and 200 mmol·L^-1 NaCl. The results showed that tetraploid wheat had the highest concentration of Na⁺, the lowest concentration of K⁺and a low K⁺ /Na⁺, diploid wheat had the highest concentration of K⁺, the lowest concentration of Na⁺and high K⁺ /Na⁺. The change of K⁺ and Na⁺ in diploid wheat was more obvious than that in tetraploid and hexaploid synthetic wheat under salinity stress. The hexaploid synthetic wheat showed a relative advantage of accumulating K⁺ compared with tetraploid wheat. Low concentration of NaCl(50 and 100 mmol·L^-1)stress stimulated the accumulation of MDA and increase of antioxidant enzyme activity in three ploidies of materials. The tetraploid wheat showed a significant superiority to accumulate osmotic adjustment substances and kept higher activity of antioxidant enzymes than diploid and hexaploid materials. Meanwhile, hexaploid synthetic wheat displayed a strong ability of accumulating soluble protein and proline as well as enhancing POD activity. Our results suggested that the diploid containing DD genome may mainly adapt to salt stress by regulating K⁺ /Na⁺ ratio while the tetraploid containing AABB genome may mainly adapt to salt stress by regulating antioxidant enzyme activity and accumulating more osmotic adjustment substances. The hexaploid synthetic wheat containing AABBDD exhibited the comprehensive adaptation mechanisms to salt stress, may express a broader adaptation ability to salt stress compared with the tetraploid and diploid parents.

Regulation of HDA19 on resistance of Arabidopsis thaliana to Botrytis cineara

2017/11/29 16:58:45
HDACs(Histone deacetylase)family proteins are responsible for the deacetylation of histones H3K4 and H4K19 and are involved in signal transduction processes of plant growth and stress. Jasmonates(JA)is an important natural plant hormone that not only regulates plant growth and development but also participates in the process of signal transduction and regulation of plant responses to multiple stress conditions. However, the specific mechanism by which HDACs participate in JA signal transduction is not very clear. We investigated the function and role of HDACs in plant JA signal transduction with HDA19(Histone deacetylase 19). In this study, we found that the HDA19 T-DNA insertion homozygous mutant did not show a significant JA root length response treated by MeJA. Under the same conditions, there was no significant difference in anthocyanin content between different mutant lines of hda19 and WT plants of the same ecotype background. However, JAZ1, VSP1 and other downstream JA signaling pathway marker genes were significantly up-regulated expression. At the same time, the resistance of hda19 to fungi Botrytis cineara was significantly enhanced compared with WT, and the expression levels of basic defense marker genes PDF1.2, Thi2.1 and ERF1 were signifi-cantly higher than WT in hda19. Based on these results, we believe that HAD19 negatively regulates Arabidopsis thaliana defense against Botrytis cineara via JA signaling pathway.

Flavonoid related compounds and their biosynthesis pathways in Linum pernne

2017/11/29 16:58:45
Flower colour is one of the most ornamental quality in ornamental plant, and flavonoid is the main pigment. In this study, the flowering stages of Linum pernne was divided into five stages. We used high performance liquid chromatography with a photodiode array detector(HPLC-PAD)and HPLC-electrospray ionization-mass spectrometry(HPLC-ESI-MS)method for qualitative and quantitative analyses of anthocyanins and the key flavonoid intermediate product in the different flower development stages. The results were as follows: L. pernne petals contained delphinidin, cyanidin and malvidin, but pelargonidin was no detected, and the main color substance was acylation of delphinidin; The highest total contents of the anthocyanins was at Stage 2. According to the end product and the intermediate product, we speculate the flavonoid biosynthesis pathways in L. pernne, in which F3'5'H leading branch pathway is dominant, and the main reason for that is F3'5'H enzyme activity is higher than that of F3'H.

Genetic diversity and fingerprints of Gleditsia sinensis germplasm resource based on SCoT

2017/11/29 16:58:45
Genetic diversity of eighteen Gleditsia sinensis germplasms was analyzed based on SCoT markers, the cluster analysis on eighteen G. sinensis germplasms was carried out by UPGMA method, and the SCoT fingerprints of eighteen G. sinensis germplasms were constructed by the polymorphic bands. The results of amplification showed that, 226 SCoT bands were obtained from fifteen primers selected from 51 SCoT primers, including 216 polymorphic bands, with a polymorphism rate of 96.61%. The average of the polymorphism information content(PIC), observed number of alleles(Na), effective number of alleles(Ne), Nei's gene diversity(H)and Shannon's information index(I)were 0.875 9, 1.964 9, 1.440 1, 0.272 6 and 0.426 1, respectively. The genetic similarity coefficients of these samples were between 0.491 4 and 0.938 1. Therefore, it indicates that there are rich genetic diversities among these materials. The results of cluster analysis showed that, eighteen G. sinensis germplasms could be divided into three groups when the genetic similarity coefficient was 0.60. G. heterophylla formed the first group, G. melanacantha and Zaojia-T formed the second group, and the others formed the third group. DNA fingerprints of eighteen samples were constructed from eight polymorphic loci amplified by three primers, and these materials could be distinguished and identified accurately. All these results provide the important information for the identification and breeding for new cultivars of G. sinensis.

Tissue culture and rapid propagation technique of the superior tree from Paulownia fortunei

2017/11/29 16:58:45
Using excellent stem segments of Paulownia fortunei as explants, tissue culture and rapid propagation technique for P. fortunei were studied in this paper. The results showed that the optimal sterilization time of 0.1% HgCl₂ was 7 m; the suitable primary induction medium was MS+ 6-BA 2.0 mg·L^-1+ IBA 0.2 mg·L^-1+ sucrose 30 g·L^-1+ agar 3.5 g·L^-1(pH 5.8), 30 d later, bud induction rate was 70%; the suitable method for multiplication culture was MS+ 6-BA 4.0 mg·L^-1+ IBA 0.4 mg·L^-1+ sucrose 30 g·L^-1+ agar 3.5 g·L^-1(pH 5.8)and MS+ 6-BA 0.4 mg·L^-1+ IBA 0.04 mg·L^-1+ sucrose 30 g·L^-1+ agar 3.5 g·L^-1(pH 5.8)alternately,health and vitrification multiple shoots with this method was lower than 5%, multiplication factor was higher than 6.0/25 d; the optimal medium for rooting was 1/2MS + NAA 0.2 mg·L^-1+ sucrose 20 g·L^-1+ carrageenan 3.4 g·L^-1(pH 5.8), 14 d later, the rooting seedlings were obtained, rooting number were 5-10, root length was 3-5 cm, the rooting rate was 98% and whose roots were white, with less and short hair, easy to clean. Through acclimatization, the rooting seedlings were transplanted in the greenhouse, 50 d later, whose average height was 1.0 m, ground diameter was 1.0-2.0 cm, and survival rate was above 90%.

Tissue culture and rapid propagation technique of red-flowered strawberry

2017/11/29 16:58:45
We used leaves of red-flowered strawberry as materials, by selecting the best medium for the callus induction, adventitious bud induction and root growth to establish a rapid propagation technique system of red-flowered strawberry. The results showed that TDZ was better than 6-BA in the induction of callus, and TDZ and NAA combination had better effects than TDZ and IBA. When the concentration of 6-BA was 0.5 mg·L^-1, the inductivity of adventitious bud reached 86.6%. The low 6-BA concentration and 8 g·L^-1 agar were benefical for seedling growth. NAA was better than IBA in the rooting inducing. So the optimum medium for callus inducing was: MS + 1.0 mg·L^-1 TDZ + 0.5 mg·L^-1NAA + 30 g·L^-1 sucrose +7 g·L^-1 agar; the optimum medium for adventitious bud inducing was MS + 0.5 mg·L^-1 6-BA + 0.1 mg·L^-1 NAA + 30 g·L^-1 sucrose + 7 g·L^-1 agar; the optimum medium for seedling growthing was MS + 0.1 mg·L^-1 6-BA + 0.1 mg·L^-1NAA + 30 g·L^-1 sucrose + 8 g·L^-1 agar; the optimum medium for rooting was MS + 0.5 mg·L^-1 NAA + 30 g·L^-1 sucrose + 8 g·L^-1 agar. The tissue culture seedlings grew well and the living ratio reached 93% after planted for 20 d. The establishment of rapid propagation technique system of red-flowered strawberry provides scientific and technological support for large-scale production of high quality red-flowered strawberry seedlings.

Cloning and expression analysis of farnesyl diphosphate synthase gene in Fritillaria cirrhosa

2018/9/30 15:33:27
In order to explore whether Fritillaria cirrhosa farnesyl diphosphate synthase gene(FcFPPS)plays some role in the metablic processes for synthesis of sterioid alkaloids and synthesis of terpenoids, in this study, Open Reading Frame(ORF)of FcFPPS was cloned via PCR method based on RNA-Seq. Some bioinformatic methods and softwares were used for the gene structure and function analysis. Quantitative real-time PCR(qRT-PCR)was used for the analysis of gene expression between wild and regeneration bulbs. The total alkaloid content of wild and regenerated bulbs were measured by decoction method. The result showed that the FPPS ORF was 1 059 bp, encoding 352 amino acids. NCBI Blast x search results showed that FcFPPS had the higher amino acids similarity to the corresponding proteins from Ilium longiflorum, Ornithogalum longebracteatum and Cymbidium goeringii. The identities of FcFPPS were more than 85%. The prediction results of FcFPPS protein secondary and tertiary structure showed that the FcFPPS protein was mainly composed of α helix. The FcFPPS qRT-PCR results were in parallel with the physiological data of accumulation of alkaloid, both showed higher alkaloid accumulation in regeneration bulbs than wild bulbs. These findings suggest FcFPPS may be a biological functional protein induced by qRT-PCR combination, which establishes a theoretical foundation for the improvement of the alkaloid content by the genetic engineering.

Isolation and characterization of microsatellite markers for Akebia trifoliata

2018/9/30 15:33:27
Akebia trifoliata is perennial, woody vine producing large edible fruits. The high medicinal and nutritional values of A. trifoliata make it worthy of being exploited as a new crop. Wild resources of these species have been seriously deteriorated due to years of disorder planting and over-harvesting, thus wild germplasm resources protection and evaluation are particularly important. In order to obtain suitable molecular markers for genetic structure and genetic diversity of the existing resources, enriched SSR library was established using the magnetic bead enrichment procedure. Seventy SSR loci were obtained in 150 positive clones, and enrichment efficiency was 46.67%. Thereinto, sequences with two-base repeat unit accounted for 79.37%, far more than sequences with three-base and four-base. Sixteen pairs of high polymorphic primers were chosen in sixty-three pairs primers and characterized by 48 individuals collected in Lushan. The allele numbers per locus ranged from 10 to 22, and the observed and expected heterozygosity ranged from 0.370 to 0.792 and from 0.724 to 0.936, respectively, polymorphisms information content ranged from 0.725 to 0.919, which showed that the above primers were high polymorphic primers. Twelve pairs of primers deviated from Hardy-Weinberg equilibrium, showing excess of homozygotes, which might be related to null genes and other reasons. These molecular markers will contribute to evaluation on genetic diversity and population structure, lay a foundation for conservation and evaluation of A. trifoliata genetic resource.

Preparation biochar of corn stover and structural characteristics analysis

2018/9/30 15:33:27
Biochar is rich in carbon, which can reduce carbon emissions greatly by carbon sequestration, and play a significant role in controlling the diffusion of pollutants and promoting plant growth. Biochar, as a by-product of a variety of agricultural waste, such as corn stove, can increase the added value of agricultural products and increase agricultural income. To resolve the problem of the processing and utilization of corn stalk in North China in a high efficient, economic and environment-friendly way, in this study, a series of biochars were made from corn stove under different temperatures(200, 300, 400, 500 ℃)using oxygen-limited pyrolysis method. The pyrolysis kinetics, structure, morphology, element composition, specific surface area, pore dimeter distribution and surface functional groups were analyzed thoroughly and systematically by according methods, respectively. The results showed the biochars prepared under different pyrolysis temperatures possessed differential pyrolysis kinetics and distinct surface functional groups, which meant the pyrolysis process of different biomasses. With the heating-up process of pyrolysis, the yield, contents of nitrogen and carbon decreased, but the hydrogen and oxygen increased; meanwhile, the ratios of H/C and(O+N)/C decreased, which meant the decreasing of hydrophilicity and polarity and the increasing of aromaticity and stability. Thermogravimetric curve and differential thermal curve included three processes respectively. When the pyrolysis temperature was high, the weight loss ratio was low and the curve tended to be gentle. The specific surface area, micropore specific surface, medium pore volume, micropore volume increased with the elevation of pyrolysis temperature, but the most probable aperture decreased; moreover, adsorption capacity enhanced. In conclusion, the biochars could be prepared under 400 ℃ with a relative high yield and the most stable structure and the best adsorption performance could be obtained

Selection and validation of reference genes for quantitative real-time PCR during seed germination of Zanthoxylum dissitum

2018/9/30 15:33:27
In order to understand the molecular mechanism of the seed germination, we need to screen the reference genes of stable expression during the seed germination of Zenthoxylum dissitum. The seeds of Z. dissitum were treated with the gibberellins(GA)to promote germination. The samples were collected in 0, 1, 2, 3 and 5 d of germination. The quantitative real-time PCR(qRT-PCR)was performed to analyze the expression stabilities of six candidates such as Glyceraldehyde-3-phosphate(GAPDH), Actin(ACT), 18S ribosomal RNA(18SrRNA), ubiquitin-40S ribosomal(UBQ5), α-tubulin(TUA) and cyclophilin(CYP)among the seeds of the different germination stages. The α-amylase gene, DELLA gene and isocitrate lyase gene respectively reflected the metabolic activities of the sugars, hormones and lipids during the germination of Z. dissitum seeds. Therefore, isocitrate lyase (Unigene0032088), α-amylase(Unigene0033597)and DELLA (Unigene0058868)of Z. dissitum were selected as the validation genes for the relative expression verification during the seed germination. The comprehensive analysis of three softwares including geNorm, NormFinder and BestKeeper indicated that the expression stablility of ACT was the best and the second was UBQ5. The expressions of the validation genes were consistent with the states of seeds germination when ACT and UBQ5 were selected as the reference genes. Besides, when the ACT and UBQ5 genes were selected as the reference genes, the relative expression levels of isocitrate lyase, α-amylase and DELLA of zanthoxylum revealed the possible reasons that the seeds treated with GA were able to germinate successfully but the seeds treated with water easily spoiled in the 3rd day of germination. In conclusion, ACT is the preferred reference gene during the seed germination of Z. dissitum, followed by UBQ5.

Cloning and expression analysis of a new reducing polyketide synthase gene in Antrodia camphorata

2018/9/30 15:33:27
We isolated the polyketide synthase gene with anlyzing the genome of Antrodia camphorata, designed special primers including initial coden and stop coden according to the DNA sequence of this gene. Then, we cloned the full length of a PKS gene using the cDNA of A. camphorata, it is a part of HR-PKS and named as AcPKS2. We used bioinformatic methods to analyze AcPKS2 gene and its proteinic sequence, and compared the expression level of AcPKS2 gene culturing in different media. The results showed that the gene AcPKS2 had 7 842 bp including 24 introns, all the exons encoded 2 613 amino acids; and its relative molecular weight was 293.5 kDa, the theoretical isoelectric point(pI)was 5.78. The result of conserved domain database(CDD)analysis revealed that the organization domain of AcPKS2 was KS-AT-DH-MT-ER-KR-ACP-TE, the active sites of eight domains were β-ketosynthase(DTACSSSL), acyltransferase(GHSIGETA), dehydratase(RNDGSTSPL), methyl-transferase(SFDIITAFDV), enoyl reductase(HAGVSSPAA), ketoreductase(GSPGQANYTAA), acyl carier protein(YGLDSLTSVRL), thioesterase(KQPNGPY), respectively. Phylogenetic analysis clarified that AcPKS2 and another HR-PKS that their products had not identified clustered together, the domain and phylogenetic tree analysis could anthenticated that AcPKS2 gene encoded a new HR-PKS that includes TE domain. The gene expression analysis showed glucose and fructose could improve the ability of AcPKS2 gene expression.

Genome MSAP analysis of Haematococcus pluvialis astaxanthin accumulation under nitrogen depletion stress

2018/9/30 15:33:27
Haematococcus pluvialis is the best resource for natural astaxanthin, which has multiple biological functions. Nitrogen depletion can result in astaxanthin accumulation in H. pluvialis. In order to explore the molecular mechanism of astaxanthin accumulation in H. pluvialis under nitrogen depletion stress, the growth rate of the algae was found to decrease during 0-72 h, the accumulation of astaxanthin mainly occured during 12-24 h, and then slowed down. MSAP analysis of stress time point at 0, 24 and 72 h got 291 methylation polymorphism loci, among which 29.90% of 0-24 h methylation loci and 53.95% of 24-72 h methylation loci changed. After 24 h of nitrogen depletion stress, the DNA semi-methylation rate was the highest(12.71%), and the full-methylation rate got the lowest(26.80%). On the contrary, full-methylation rate was the highest(28.52%)and the semi-methylation rate was the lowest(1.72%)after 72 h stress. DNA methylation changes appeared to be a vital regulation for astaxanthin accumulation.

Effects of phytagel and sucrose on regeneration efficiency of somatic embryos and growth of regenerated plants in Hevea brasiliensis

2018/9/30 15:33:27
Phytagel and sucrose have great influence on the regeneration efficiency of somatic embryos and growth of regenerated plants. In this study, effects of phytagel and sucrose on the regeneration of somatic embryos and growth of regenerated plants were carried out on mature cotyledonary embryos from Hevea brasiliensis clone Reyan7-33-97. MS me-dium supplemented with 0.23 μmol·L^-1 KT, 0.11 μmol·L^-1 IAA and 8.7 μmol·L^-1 GA3 was used as the plant regeneration medium. The results showed that appropriate phytagel was helpful for the regeneration efficiency of somatic embryos and growth of regenerated plants. At lower concentrations(0-1 g·L^-1), the frequency of plant regeneration increased with the increase of the amount of phytagel, but at higher concentrations(1-4 g·L^-1), plant growth was inhibited along with the increasing amounts of phytagel. When the phytagel was supplemented in somatic embryo regeneration medium with 1 g·L^-1, the plant regeneration rate, the plants(height >5 cm)and the plants with leaves reached(86.4 ± 5.7)%,(53 ± 9.4)% and(81.7 ± 3)%, respectively, higher than other treatments; For sucrose, no effect on the regeneration efficiency of somatic embryos was observed when sucrose was supplemented in somatic embryo regeneration medium, however, significant effects on growth of regenerated plant were observed. When the sucrose was supplemented in somatic embryo regeneration medium with 50 g·L^-1, the plants(height >5 cm)and the plants with leaves(height >5 cm)reached(57.6±5.4)% and(46.3±12.3)%, respectively, higher than other treatments, and the stems and roots were very strong from the outside. Moreover, lower sucrose(20-30 g·L^-1)promoted the growth of leaves as plant sprouting, but the elongation of stem was supressed, in contrast, higher sucrose(70-80 g·L^-1)suppressed the growth of leaves but promoted the elongation of the stem. Overall, the somatic embryo regenaration medium supplimented with 1 g·L^-1 phytagel and 50 g·L^-1 sucrose was the most approciate for somatic embryo regenaration in H. brasiliensis.

Microtuber induction of potato cultivar ‘Mira' by “solid + liquid double-layer” culture

FANG Zhirong, ZHOU Caiyi, LI Peihua, QING Yuan — 2018/9/30 15:33:27
Virus-free plantlets of potato cultivar ‘Mira' were used as experimental material to induce potato microtuber by a two-step culture method. In the first step, nodal cuttings(with one or two leaf)of potato plantlets were inoculated solid medium for culturing vigorous potato plantlats. After 20 d, liquid medium was added on the solid medium to proceed to the second step for induction of potato microtuber. The optimum culture media at the stages of culturing vigorous potato plantlets and induction of potato microtuber were obtained by means of orthogonal design, and the effects of sucrose concentration, light condition and CCC concentration on induction of potato microtuber were also studied. The results were as follows: the optimal medium at the stage of culturing vigorous potato plantlets was modified by MS medium(NH₄NO₃ was changed to 2 000 mg·L^-1 and KNO₃ was changed to 2 000 mg·L^-1 in MS medium)+ 500 mg·L^-1 CCC + 0.1% activated carbon + 0.1 mg·L^-1 DA-6 + 1 mg·L^-1 6-BA + 0.1 mg·L^-1 NAA + 3% sucrose + 6 g·L^-1 agar, pH 5.8, the optimal medium at the stage of induction of potato microtuber was MS₁ medium [two times of FeSO₄(Na₂EDTA)and trace elements in MS medium] + 1.5% activated carbon + 4 mg·L^-1 6-BA + 8% sucrose. The results also showed that illumination with dim light for 4 h·d^-1 could obtain potato microtubers with higher frequency of tuberization and big microtubers than dark treatment. The results indicated that“solid + liquid” double-layer culture can be applied for mass propagation of potato tubers ‘Mira' at low cost in the plant tissue culture room, and can increase the number of pre-elite seed potatoes. This method can also be used for the production of microtuber of other potato cultivars.

Establishment of somatic embryogenesis system of Lycium barbarum ‘Ningqi 8'

2018/9/30 15:33:27
In order to establish a high-frequency regeneration system for Lycium barbarum. The leaf of L. barbarum ‘Ningqi-8' was selected as material to explore the crucial effects of hormone combinations and additives on the induction of somatic embryogenesis, somatic embryo proliferation and germination and plant regeneration. The results showed that by orthogonal analysis of 6-BA, 2,4-D and IAA, the optimal hormone combination of L. barbarum ‘Ningqi-8' somatic embryos was 6-BA 1.0 mg·L^-1+2,4-D 0.3 mg·L^-1+IAA 0.4 mg·L^-1, the highest induction rate could get to 88.67%. Range analysis showed that 6-BA had the most significant effect on somatic embryogenesis. Using suitable proportion of hormone combination was possible to induce somatic embryogenesis of L. barbarum ‘Ningqi-8', which had a large number of normal morphology. In the culture of somatic embryos, the high concentration of 6-BA was easy to cause vitrification, which was not conducive to the growth and proliferation of L. barbarum ‘Ningqi-8' somatic embryos. The proliferation rate of somatic embryos increased with the increase of hormone concentration, and the vitrification rate increased, too. Then the most efficient proliferation medium for L. barbarum ‘Ningqi-8' was 6-BA 0.4 mg·L^-1+NAA 0.6 mg·L^-1, and the highest germination rate of L. barbarum ‘Ningqi-8' somatic embryo was obtained when IBA 0.3 mg·L^-1+GA₃ 0.4 mg·L^-1+ sucrose 10 g·L^-1 was added. The highest germination rate could get to 89.17%, and the degree of influence on the germination of L. barbarum ‘Ningqi-8' was IBA > sucrose > GA₃. Adding GA₃ and low concentration sucrose could promote the mature somatic embryo germination. Activated carbon could effectively improve the regeneration rate of L. barbarum ‘Ningqi-8' somatic embryo, and simultaneously promote the development of the root of germination embryo. IBA 0.1 mg·L^-1+KT 0.4 mg·L^-1+ activated carbon 1 g·L^-1 was found to be the suitable medium for regeneration in which L. barbarum ‘Ningqi-8' demonstrated the highest multiplying capacity, and the regeneration rate could get to 91.67%.

Cloning and expression analysis of small heat shock protein gene BsHsp17.3 in Bletilla striata

2018/9/30 15:33:27
BsHsp17.3 gene was isolated from Bletilla striata using homologous cloning and 3'RACE methods. In this study, the expression profiles of BsHsp17.3 under different stresses were analyzed and suitable conditions were explored for artificial cultivation of B. striata. The results were as follows: The opening reading frame of BsHsp17.3 was 453 bp, which encoded a 150-amino acid peptide. Its protein molecular weight and isoelectric point were 17.42 kD and 6.33. Phylogenetic analysis demonstrated that BsHsp17.3 was closed to the Hsp17.3 from Dendrobium catenatum. Quantitative real-time PCR analysis showed that the expression of BsHsp17.3 was different in leaf, root, bulb and flower of B. striata. The expression level of BsHsp17.3 gene was the highest in leaf and root,respectively, but was absent from the bulb and flower. The expression of BsHsp17.3 was analyzed by quantity RT-PCR under cold, hot and drought treatments. The results showed that the expression of BsHsp17.3 quickly induced by high temperature and cold, but drought stress simulated by 20% PEG did not regulate the gene expression. These results suggest that BsHsp17.3 may be involved in regulating sprout tumble of Bletilla striata.

Cloning,bioinformatic and expression analysis of LaAP2 gene from Lepidium apetalum

2018/6/26 11:36:15
AP2/ERF is a kind of important transcription factors widely existing in plants. AP2/ERF regulate the expression of some genes involved in abiotic stress, and help plants improve the ability of adversity stress. We obtained a gene LaAP2 by cloning a significantly up-regulated expression AP2/ERF family gene sequence by RT-PCR technology, based on the previous analysis of Lepidium apetalum transcriptome database. Bioinformatics analysis of LaAP2 encoded amino acid sequence composition, physical and chemical properties, structure, hydrophobicity, transmembrane region, signal peptide, subcellular localization and secondary protein structure. Real-time PCR was used to detect the expression of LaAP2 gene at 4 ℃ low temperature stress. The result showed that the full-length cDNA of the gene was 1 005 bp, and the encoded amino acid sequence contains a AP2 and a B3 domain, belonging to the AP2/ERF transcription factor RAV subfamily. The mole-cular weight of putative LaAP2 protein was 37.744 67 kD and the isoelectric point was 9.49. The amino acid sequence has high homology with Camelina sativa, Arabidopsis thaliana and rape. Phylogenetic analysis showed that the genetic relationship was close to that of A. thaliana. The amino acid sequence analysis showed that LaAP2 gene encoding protein had no signal peptide segments, no transmembrane region, did not belong to the secreted protein. It could be the hydrophilic protein. The possibility of localization in cytoplasm was 56.5%, and in nucleus was 21.7%. The main structural elements were random coil, extending strand and alpha helix. We analyzed the expression of AP2/ERF gene in Lepidium apetalum seedlings under 4 ℃ low temperature stress through Real-time PCR technology, which showed that LaAP2 gene expression was decreased firstly and then increased. The results suggest that LaAP2 play a role in regulating the resistance of low temperature stress in Lepidieae seedling. This study lays the foundation for further study on the function of LaAP2 on the L. apetalum seed germination and seedling growth in tolerance to low temperature.

Screening of interaction proteins of GS3 in rice using BioID

2018/6/26 11:36:15
GS3, encoding the noncanonical γ subunit of heterotrimeric G proteins system in rice, is a major quantitative trait locus for grain size in different rice varieties, serving as a negative regulator in seed or organ size. Newly-developed BioID(proximity-dependent biotin identification)system, which detects proximal proteins in vivo, has been successfully applied in different species for its features of sensitivity, high efficiency and fast speed. And the mechanism of BioID is that the biotin ligase, the core component, could attach biotin to the proximal proteins, and the biotin also connects tightly with streptavidin, therefore, the beads coupled streptavidin could be utilized for enriching the target proteins. In this study, we screened proteins interact directly or indirectly with GS3 in rice protoplasts by BioID method. The results of Western-blot showed that fused protein BirAG-GS3 expressed and biotinylated the proximal proteins of GS3 in the protoplasts successfully. Biotinylated proteins were enriched by beads coupled streptavidin and the Mass Spectrometry sequencing performed on it. Then, the function of proteins has been annotated and enriched, meanwhile, protein-protein interaction graph was established. Some of the candidate proteins were confirmed by BiFC, indicating the possible interaction between ICL, PPDK, RPN7 and RH15, and the engagement of GS3 in regulation of energy metabolism, stock of starch, ubiquitin-proteasome system and apoptotic pathway. The above results will facilitate the elucidation of the regulation network of the function of GS3.

Influence of AgNO₃ on tropane alkaloids and key enzyme genes in hairy roots of Atropa belladonna

2018/6/26 11:36:16
In this study, we studied the influences of different concentrations of AgNO₃ on tropane alkaloids and key enzyme genes in metabolic pathway. We all know that the accumulation of plant secondary metabolites is directly related to some tprimary metabolism and activity of key enzymes. And AgNO₃ is a heavy metal, which can induce oxidative stress in plants and which has a great effect on plant growth and key enzymes of metabolism. Therefore, we added five concentrations of AgNO₃ into Atropa belladonna hairy roots, and cultivated in B5 liquid medium for 12 d, and then, we collec-ted hairy roots and determined fresh weight, dry weight, contents of tropane alkaloids, some physiological indexes included MDA, Pro, soluble sugar and soluble protein, genes expression level after cultivated for 2 d. The results showed that AgNO₃ promoted the accumulation of the alkaloids, although AgNO₃ inhibited the growth of hairy roots, 50, 100, 150 μmol·L^-1 AgNO₃ could improve the contents of tropane alkaloids compared with control. Meanwhile, 150 μmol·L^-1treatment group significantly improved the hairy roots of the content of MDA compared with control. As well as the content of Pro increased to 5.92 μg·g^-1FM, 2.91 times of control. Then, the contents of soluble sugar and souble protein were significantly improved 1.55 and 1.67 times of control respectively by treating with 150 μmol·L^-1 AgNO₃ treatment. Thus, with the treatment of AgNO₃, gene expressions(pmt, trI, h6h) of key enzymes in the metabolic pathway of A. belladonna hairy roots all increased their produced effects. Therefore, we could be inferred that the contents of tropine alkaloids in A. belladonna hairy roots could regulate and control primary metabolism such as Pro, soluble protein, soluble sugar and secondary metabolism like expressions of key enzyme genes with the treating of AgNO₃. This study could provide the basic theories for study the synthesis mechanism of tropane alkaloids and an effective method to enhance the medicinal composition in the culture of hairy roots of Atropa belladonna in the future.

Selection of reference genes for quantitative real-time PCR during anther development of Caulokaempferia coenobialis (Zingiberaceae)

2018/2/6 10:48:17
Quantitative real-time PCR(qRT-PCR)has been widely used in gene expression analysis with quantitative accuracy, high sensitivity and good repeat ability. The stability of the reference genes is very important to accurate analysis of the target genes expression. Anther tissues from pollen mother cells stage(PMC), tetrad stage(TET)and mature pollen stage(MP)of Caulokaempferia coenobialis (Zingiberaceae)were used to select reference genes. According to the developing anther transcriptome expression profile data and traditional reference genes reported, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), Malate dehydrogenase (MDH), alpha tubulin3 (TUA3), beta tubulin7 (TUB7) and Actin6 (ACT6)were selected as candidate reference genes for qRT-PCR analysis. The expression stability of five candidate reference genes during the anther development was comprehensive analysis by BestKeeper, geNorm and Normfinder software. The results showed that the expression of MDH and TUB7 were the most stable, and the ACT6 was the worst. The expression patterns of GBE1 during anther development was obtained based on MDH and TUB7 as reference gene respectively. And the expression stability of MDH and TUB7 was further verified by correlation coefficient analysis(Pearson)with the anther transcriptome expression profile data. The results showed that MDH and TUB7 could serve as qRT-PCR reference gene to analyse the gene expression pattern related to anther developing in C. coenobialis. The study would provide research fundamental data for molecular mechanism research of anther development in C. coenobialis, and reference for the selection of reference genes in other Zingiberaceae species during anther development.

Clone and sequence analysis on gene BnFY34 from spring rapeseed(Brassica napus)

2018/2/6 10:48:17
Rape, belonging to Brassica(Cruciferae), is an important oil crop widely cultivated in China due to its wide adaptability. Spring rapeseed(Brassica napus)is widely cultivated in Qinghai Province due to its high yield and quality, however, it has been greatly restricted the promotion of rape in alpine region because of its long development period. Therefore, screening differentially expressed genes in seedling and bud stages plays an important role in early seedling breeding of spring rape with high yield and quality. In this study, a gene, named BnFY34, specifically expressed gene under seedling stage, was screened out with cDNA-AFLP, and successfully cloned in full-length 455 bp with RACE, including complete Open Reading Frame(ORF)and coding a protein with 71 amino acid residues. The protein has a molecular weight of 8.04 kD and a theoretical isoelectric point of 10.25. Furthermore, three α-helix and no β folding structures existed in the tertiary structure of protein BnFY34. The BnFY34 gene was compared with the genome sequence of Brassica napus. The results showed that the BnFY34 gene located in the C4 chromosome of B. napus and consisted of three exons and two introns. In addition, the genetic relationship between the gene BnFY34 and homologous genes in other plants showed that the gene BnFY34 was under the same subfamily as the Brassica species, and was similar to that of the untranslated gene XM_013837282.1 reported in B. napus up to 100%, but its function needs to be further studied.

Transcriptome-based excavation and analysis of MYB family transcription factors in Phoebe zhennan

2018/2/6 10:48:17
Phoebe zhennan is an endemic valuable tree to China and listed as one of the national second-class protected wild plants. The growth rate of P. zhennan is too slow and the operating cycle is too long. To date, there is no reason to explain this phenomenon. These years have witnessed transcription factors becoming a focus on the researches about plants molecular biology, as the high-throughput sequencing technology has been well applied and developed. In the present study, the identification and phylogenetic analysis of the P. zhennan MYB(PzMYB)transcription factor family was performed and the expression profiles of these genes were determined. PzMYBs were identified from the P. zhennan transcriptome using bioinformatics tools, and their putative functions were determined based on the phylogenetic tree and classified into subfamilies using AtMYBs describing known functions. The results showed that 82 PzMYB transcription factors were excavated, and they encoded 50-1 121 amino acids, predicting hydrophilic and unstable. Their main structural elements were α-helix and irregular curl. Also, the sequence analysis and construction of phylogenetic tree showed that these MYB transcription factors had a certain conserved type, containing [W]-X(19)-[W]-X(19). Eighty-two PzMYB could be divided into 22 categories, and were related to the plant response to biotic stresses, cell development, secondary metabolism etc. Their involvements in response to stresses were reported by several transcriptional studies, which was precisely consistent with the functional prediction. Lastly, the members in the same subfamily had different spatial and temporal expression profiles, with genes in stems and leaves expressed at various levels, which was precisely consistent with the functional prediction. In stem, the numbers of differential expression of genes were eighteen, ten up-regulated and eight down-regulated in leaf. Based on the transcriptome data of P. zhennan, an endemic tree in China, we excavated and analyzed its MYB transcription factors, and provided valuable information for PzMYB gene cloning and functional characterization of P. zhennan. This research will lay a foundation for the study of molecular biology of P. zhennan and provide references for its genetic modification and molecular breeding.

Over-expression of cytosolic copper/zinc superoxide dismutase gene and increase cold-tolerance in Eucalyptus grandis × E. ophylla

ZHAO Yanling, HAN Yao, JIN Yulei — 2018/2/6 0:00:00
Copper/zinc superoxide dismutases are important antioxidant enzymes to guard cells against superoxide toxicity. The over-expression of CuZnSOD can improve the resistance and recovery ability of plants to abiotic stresses such as salt, drought, cold and high temperature stress. The plant genome encodes three types of CuZnSOD, the CuZnSOD1 in cytoplasm, the CuZnSOD2 in chloroplast and the CuZnSOD3 in peroxisome or extracellular. The cytosolic EuCuZnSOD1 gene was cloned from Eucalyptus grandis &#215; E. ophylla(GenBank Accession Number: JX138573)and the biological function was verified by prokaryotic expression system. The fusion vector of sense EuCuZnSOD1 gene derived by CaMV35S promoter was constructed and transformed to Eucalyptus grandis x 215; E. ophylla by Agrobacterium. Using Kanamycin selection and PCR analysis, ten transgenic eucalyptus lines were obtained. Compared with the control plants the tissue culture transgenic eucalyptus No. 40 and No. 41 could be well tolerant in -13.1 ℃ for 5 h and recovery without obvious damage. There was no distinct increase in SOD enzyme activity in transgenic plant paralleled with wild plant under room temperature and 4 ℃ for 48 h. However, the real-time PCR analysis showed that the expression of EuCuZnSOD1 was raised to about nine times higher than wild line under room temperature. After 4 ℃ for 36 h, the expression of EuCuZnSOD1 in No. 40 and No. 41 line was respectively increased about 16 and 36 times higher than the wild plant and then down to the normal level at 4 ℃ for 48 h. The data indicated that the increased expression of EuCuZnSOD1 improves the cold tolerance of Eucalyptus. The klason lignin content of transgenic Eucalyptus was reduced especially the No. 41 Eucalyptus was decreased to 56% of the control plant except for the No. 40 plant was no significantly different from the control plant. In conclusion, over-expression of the cytosolic EuCuZnSOD1 in Eucalyptus grandis &#215; E. ophylla could enhance the cold-tolerance and improve the recovery ablity.

Isolation,identification and inhibiting effects of pathogens that caused two kinds of diseases in Lilium brownii var. viridulum

2018/2/6 10:48:17
The pathogens that caused two kinds of diseases in Lilium brownii var. viridulum were isolated using usual tissue isolation. Strains were cultivated, purified, reinoculated and reisolated. Morphological and molecular biological techniques were used to identify the strains. And three plant growth regulators were used to study the inhibiting effects on two pathogens. The results showed that two kinds of diseases were leaf blight(A)and penicilliosis(B)by the symptoms, and four strains and three strains were isolated from infected leaves, respectively. But only A-4 and B-2 caused two kinds of diseases, respectively. The incidences of A-4 and B-2 disease reinoculated in the field were all 100%. A-4 is a kind of fungi belonging to the genus Botrytis, and its colonial morphology is a circular form, with colorless mycelium, prostrate on the medium, aerial, diaphragms and branches. There are conidium and acrospore. B-2 is a kind of fungi belonging to the genus Penicillium, and its colonial morphology is a circular form or irregular, with white mycelium outside the circle and turquoise in the central, the mycelium prostrate on the medium and diaphragms. There were broom conidium and spherical spores. At last, the sequence of internal transcribed spacer(ITS)region of A-4 and B-2 were analyzed, the length was 522 bp and 551 bp, respectively. The sequence were compared with other species in the Genbank, respectively. The symptom in the field indicates that the pathogen of Lilium brownii var. viridulum leaf blight(A)and penicilliosis are Botrytis elliptica and Penicillium expansum. The inhibiting effects of three plant growth regulators on two pathogens indicated that two pathogens could not be fully inhibited by 0.1-1.0 mmol·L^-1 SA, but could be fully inhibited by 0.5-1.0 mmol·L^-1BRs or Me-JA.

Gallic acid isolated from Sedum lineare and its control efficacy on rice bacterial leaf streak

2018/2/6 10:48:17
A compound, which could significantly inhibit the growth of Xanthomonas oryzae pv. oryzicola(Xoc)in vitro, was isolated from Sedum lineare by liquid-liquid extraction, silica gel column chromatography and gel column chromatography. Based on the spectral data, including ¹H and ¹³C NMR data, this compound was identified as 3,4,5-Trihydroxybenzoic acid, namely gallic acid(GA). The results of the test, on determining the antibiotic spectrum of GA at concentration 30 mg·mL^-1, indicated that GA could most strongly inhibit the growth of X.campestris pv. pruni, X. oryzae pv. oryzicola, X. oryzae pv. oryzae and X. axonopodis pv. citri, with inhibition zones width of 25.33, 24.00, 22.33 and 20.67 mm, respectively, and had fair anti-bacteria activity against Ralstonia Solanacearum, Pseudomonas syringae pv. glycinea, P. syringae pv. tomato and Pectobacterium carotovora subsp. carotovora, with inhibition zones width of 18.00, 12.33, 11.00 and 8.68 mm, respectively. Meanwhile, GA could weakly inhibit the growth of eleven pathogenic fungi, including Phytophthora nicotianae, Penicillium digitatum, Streptobotrys streptothrix, Pythium aphanidermatum, Pestalotiopsis mangiferae, Curvularia lunata, Rhizoctonia solani, Fusarium oxysporum f. sp. niverum, F. oxysporum f. sp. nicotianae, Botrytis cinerea and Sclerotium rolfsii, with inhibition ratio of 56.34%, 54.73%, 38.00%, 29.86, 28.17%, 20.00%, 19.02%, 16.71%, 10.59%, 4.58% and 1.96%, respectively. Growth inhibition exceeding 50% by GA was only observed in Phytophthora nicotianae and Penicillium digitatum. The growth of Botrytis cinerea and Sclerotium rolfsii was slightly inhibited by GA. In potted experiment, gallic acid at 200, 300 and 400 mg·L^-1 provided 63.55%, 71.38% and 77.39% control efficacy to rice bacterial leaf streak, respectively. At concentration of 300 mg·mL^-1, GA could control rice bacterial leaf streak, with control efficacy of 64.62% in field. The study reveals that the gallic acid has the potential to be further developed as a bactericide to control rice bacterial leaf streak.

Techniques for rapid propagation of Arundo donax

2018/2/6 10:48:17
In order to find out the most efficient way for rapid propagation of Arundo donax, replanted at Guangxi Insti-tute of Botany from Hungary, by testing different concentrations of plant growth regulators, and stems with auxiliary buds were used as explants. The results showed that the most suitable medium for the sprouting of auxiliary buds was MS+ 6-BA 4.0 mg·L^-1+sucrose 30 g·L^-1 +agar 3.5 g·L^-1, the germination rate could get to 90.0%. MS+ 6-BA 8 mg·L^-1+ IBA 0.8 mg·L^-1+ sucrose 30 g·L^-1+ agar 3.5 g·L^-1was the most efficient proliferation medium, and buds were robust. The average proliferation rate was 4.3/30 d. Three or five sprouts linked together was found to be a better basic material for subculture in that they demonstrated a higher multiplying capacity. The most suitable rooting medium was 1/2MS+ NAA 0.2 mg·L^-1+ sucrose 20 g·L^-1+ activated carbon 1 g·L^-1with the five cm buds, in which the rate of rooting was 100% for 7 d, the average root number was four, and the length of the roots was 2-4 cm. The rooted seedings, which with roots 2-3 cm, were transplanted from culturing seedlings to fine river sand, yellow soil or humus soil after one week-day's treatment, the survival percent reaches 98% after a month. The current annual yield was about 5 500 kg on one acre by regular management in the experimental field. The research provides technical support for industry application of the improved Arundo donax preliminarily.

Photoautotrophic rooting of sugarcane microshoots

2018/10/28 18:24:41
The purpose of the study on photoautotrophic rooting of sugarcane microshoots was to simplify the process of sugarcane tissue culture and reduce the production cost. In this experiment, the sugarcane variety GT 44 and B9 rootless microshoots, were used as test materials. The microshoots were first treated with leaves and sprayed with plant growth regulator and then hardened for 24 hours. Then the treated microshoots were transplanted into mixture of soil and sand, the adventitious root formation and growth process were completed under sunlight greenhouse conditions. Meanwhile, the survival rate and growth of rootless and rooted microshoots were compared during transplanting in the green house. The rooting rate of microshoots was measured from the third day after microshoots transplantation to the end of the tenth day. The investigation of survival rate was on the 30th day after microshoots transplantation. The results showed that the survival rate of rootless microshoots treated with indole butyric acid(IBA)and ABT2 rooting powder was to 96.3% and 97.7%, respectively, which was close to the survival rate of traditional rooting microshoots. The rooting cost of microshoots by the developed root technique was only 1/28 of that by the traditional rooting method. The first visible roots of sugarcane variety GT44 and B9 were appeared on the fourth day after transplanting of the microshoots. The regeneration of microshoot roots could be done in the sandy soil substrate and solar greenhouse instead of rooting in sterile MS rooting medium and culture room. The genotype and the quality of microshoots were the factors that affect photosynthetic autotrophic root of the key factors. The photoautorophic rooting of microshoots present more advantages over the traditional rooting technique such as simple operation, simplified protocol, the high rooting rate and survival rate, labor saving, energy saving and lower production costs and high efficiency, and it can replace the traditional root technology for commercial production.

Optimization of protoplasts preparation, culture and fusion in Pogostemon cablin

2018/10/28 18:24:41
For establishing a high efficient and stable protoplast culture and fusion technology system, the protoplasts obtained from Pogostemon cablin's callus, were studied as the material for enzymatic hydrolysis conditions of protoplast preparation, and on this base, the effects of culture methods, cell density, hormone types and concentrations on the protoplast culture were studied. Screening range of fusion cells was determined by measuring the diameter of fusion products. The effects of PEG concentration, cell density, incubation time and amount of fusion fluid on the fusion rate of protoplasts were studied by single factor analysis. The results showed that the vitality of protoplast reached the highest value at 25 ℃ and the optimal pH for enzymatically hydrolyzing was 5.8. The improved medium MS₁, which consisted half of ammonium salt(NH₄NO₃ 825 mg·L^-1), added the hormones about 0.2 mg·L^-1 NAA and 2.0 mg·L^-16-BA was used to embed in sodium alginate. With the conditions of 2.0×10⁵protoplasts·mL^-1 of the cell density, 1% sucrose content and 500 mg·L^-1 acid hydrolyzed casein, protoplasts got into division and formation of cell clusters earlier and had the higher division frequency and plating efficiency. The range of double cell fusion products was 69.33-87.35 μm; 40% PEG 6000 was selected to promote the chemical fusion. With the increase of fusion time, cell density and amount of fusion liquid, both polymerization rate and polycondensation rate increased. With 0.5 times volume of the fusion solution, 2.0 × 10⁵protoplasts·mL^-1of the cell density and 30 min of fusion time, the polymerization rate was up to 57.19%. After two months' embedding cultivation, the callus was observed.

Influences of different sodium selenite concentrations on seed germination of Metasequoia glyptostroboides

2018/10/28 18:24:41
Selenium is a trace element needed for plant growth. Enshi in which the stereoscopic natural selenium resources have been formed is the main growing place for the seed trees of Metasequoia glyptostroboides. M. glyptostroboides communities in this area are difficult to regenerate naturally, and seedlings or saplings are rarely seen under the forest. Therefore, studying the relationship between selenium and M. glyptostroboides seed germination under this environment will have great significance to explore the effects of selenium on seed germination of M. glyptostroboides combining selenium resources in Enshi. According to this ecological significance, the germination rate of M. glyptostroboides seeds was determined under different environmental conditions(20, 25, 30 ℃; 12 h Light/12 h dark cycle; pre-soaking or no-soaking), and the optimum germination conditions were selected. Under this selected conditions, different concentrations of sodium selenite(0, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg·L^-1)were used to soak the seeds of M. glyptostroboides and their germination changes were observed. The results showed that the highest value of the average germination rate(34.0%), average germination energy(29.0%)and average germination index(13.9)of M. glyptostroboides seeds were appeared at the sodium selenite concentration of 0.25 mg·L^-1. The germination rate, germination energy and germination index of M. glyptostroboides seeds were decreased with the increase of sodium selenite concentration when the concentration higher than 0.25 mg·L^-1, and the values got the lowest at 16.0 mg·L^-1, which were 0.5%, 0% and 0.025, respectively. It can be proved that the low concentration of sodium selenite(0-0.25 mg·L^-1)can promote the germination of M. glyptostroboides seeds, while the treatment with high concentration of sodium selenite(>0.25 mg·L^-1)has some certain inhibitory effects on the germination of M. glyptostroboides seeds. The findings will prove some theoretical support for field nursery and natural regeneration of M. glyptostroboides.

Oxidation reduction potential, ethylene biosynthesis, and enzyme activity of post-harvest persimmon

2018/10/28 18:24:41
Ethylene is a common plant hormone which can accelerate the ripening of fruits. Post-harvest ripening involves a variety of complex metabolic processes such as ethylene biosynthesis, reactive oxygen species biosynthesis and cell wall degradation hydrolyses. ‘Gongcheng' persimmon is a distinctive variety of Guangxi Zhuang Autonomous Region, and was used as the research material to determine the changes of indexes e.g., oxidation reduction potential(ORP), ethylene biosynthesis, firmness, total chromatism, and related enzyme activities during the storing experiment. The results demonstrated that firmness decreased continuously during the whole storing period, the firmness of fruits were treated with ethephon were declining rapidly during the first three days of storage, and were significantly lower than the control at all times. During the first fourteen days of storage, the total color chromatism was rising slowly. Fifteen days after harvest, the color of fruits, which were treated with ethephon, had changed completely and delta E value rapidly increased to the maximum of 29.6 which is evidently higher than that of the control 11.9. The ORP changed within a smaller range and maintained at 7.5 mv·g^-1in the prophase of storage. Fifteen days after harvest, the ORP of fruits which were treated with ethephon increased rapidly to 11.9 mv·g^-1, 1.4 times as much as that of the control. Additionally, ethylene biosynthesis did not change significantly during the first fourteen days. However, from the fifteenth days after harvest, the ethylene biosynthesis rate of fruits were treated with ethephon increased sharply to 0.372 μL·kg^-1·h^-1and reached the maximum, but the control kept at a stable level of 0.033 μL·kg^-1·h^-1. Thus, the increase of ethylene biosynthesis was closely related to the rise of ORP. Furthermore, the activity of peroxidase(POD)belonging to the antioxidative enzymes and β-D-galactosidase(β-D-Gal)belonging to the cell wall degrading enzymes were also significantly and positively correlated to ORP, indicating that POD and β-D-Gal activities are affected by the ORP. Therefore, the ORP of persimmon fruits treated with ethephon increased observably at the end of storage, promoting a sharp biosynthesize of ethylene, increasing activities of POD and β-D-Gal, and finally resulting in the rapid softening and ripening of persimmon. It is proved that persimmon fruit ORP may act as a switch to control the biochemical reaction of ethylene biosynthesis, and breaking the ORP steady state can initiate ethylene synthesis.

Identification and expression analysis of long chain fatty aycl-CoA synthetase gene 9 from Cinnamomum camphora

2018/10/28 18:24:41
In this study, the cDNA encoding AtLACS9 homologous gene was identified and cloned based on transcriptomes data of Cinnamomum camphora, sharing 75% sequence similarity to AtLACS9 and being registered to CcLACS9. Multiple sequences alignment showed CcLACS9 possessed three plant LACS-specific motifs and plastidical targeting signal in N-terminal. Using oleic acid as a substrate, CcLACS9 could activate free fatty acids into acyl-CoA thioesters in a yeast mutant deficient in LACS complementation test. The tissue-specific expression profile further revealed that CcLACS9 was predominantly expressed in developing seeds and flowers, but fewer in leaf and stem by real-time quantitative PCR analysis. There were 17.74-folds relative quantitative expression of CcLACS9 in kernels relative to roots. In an effort to better understand whether CcLACS9 involved seed oil biosynthesis in camphor kernel, the correlation between expression of CcLACS9 and seed oil content was surveyed. The seeds of thirty adult camphor trees were randomly sampled in November and thousand seed weight, seed oil content and the percentage of decanoic acid and lauric acid were tested and counted. According to seed oil content, the 30 individuals were subdivided into three groups and three representative plants were selected from each group to be subjected to association analysis between seed oil yield and CcLACS9 expression level in developing kernel. The results of three groups all showed that both the contents of seed oil and expression level of CcLACS9 continued to rise from June to August, and there were significant positive correlations between them. The peak of expression level of CcLACS9 in kernel was found in August. After September, the contents of seed oil tended to be stable, however, CcLACS9 keeped high expression level and had no correlation with contents of seed oil. In plant, the homoeologous genes of LACSs subfamily tend to carry the common function. These resluts implies that CcLACS9 possibly play an important role in seed oil accumulation in kernel of camphor tree.

Codon usage bias analysis of chloroplast genome of camphora tree(Cinnamomum camphora)

2018/10/28 18:24:41
In order to analyze the codon usage patterns of the chloroplast genome of camphora tree, 53 CDS(coding DNA sequences)were selected from the chloroplast genome of camphora tree and the codon usage pattern and bias were analyzed by CodonW, EMBOSS, R language and other softwares. The results showed that the effective codon number(ENC)of the chloroplast gene ranged from 36.82 to 59.30, indicating that bias of codons was weak. There were 32 codons with relative codon usage greater than 1, of which 28 were rich in A and U, indicating that the 3rd position of codon prefers A and U genetic bases. Neutral analysis found that the correlation between GC₃ and GC₁₂ was not significant, and the slope of the regression curve was 0.049, suggesting that codon bias was mainly affected by natural selection, while the ENC-plot analysis found that most genes fell below and around the curve. It was shown that the mutation also affected the formation of codon bias. Eventually,nine codons(UUU, CUU, UCA, ACA, UAU, AAU, GAU, UGA, GGA)were identified as the optimal codons for the chloroplast genome of camphora tree.

Cloning and expression analysis of IbGL3 in Ipomoea batatas

2018/10/28 18:24:41
Anthocyanin-rich purple sweet potatos have high edible and medicinal values. Anthocyanin biosynthesis is controlled by structural genes and regulatory genes. The basic helix-loop-helix protein(bHLH )transcription factor plays an important role in anthocyanin biosynthesis by regulating multiple structural genes. However, there are no reports about bHLH regulating anthocyanin biosynthesis in sweet potato. In order to further understand the function and molecular mechanism of IbGL3 in the biosynthesis of anthocyanins in sweet potato, in this study, a bHLH gene named IbGL3 was cloned in Ipomoea batatas based on transcriptome data and RT-PCR technology. The full-length cDNA of IbGL3 was 2 120 bp, containing 1 878 bp opening reading frame(ORF), and encoding 625 amino acids. The encoded protein of IbGL3 has a molecular weight of 69.08 kD and the theoretical isoelectric point(pI)of 5.20. The IbGL3 protein had highly conserved MIR motif, bHLH domain and ACT domain, shared high identities and similar domains with bHLH proteins involved in anthocyanin biosynthesis from other plants. Phylogenetic analysis showed that IbGL3 was clustered in the Ⅲ f bHLH subgroup together with other anthocyanin-related bHLH proteins. The expression of IbGL3 in the storage root of different sweet potato varieties was detected by quantitative polymerase chain reaction(qPCR). The results indicated that IbGL3 was mainly expressed in purple-fleshed sweet potato, followed by light purple-fleshed sweet potato and weakly expressed in white-fleshed sweet potato, and its expression was positively related to the accumulation of anthocyanin Ipomoea batatas. The results showed that IbGL3 may be involved in regulating anthocyanin biosynthesis in Ipomoea batatas.

-->Plant Physiology and Molecular Biology

Effects of Cr 6+on the growth and some physiological characters of tobacco by tissue culture

A genetic transformation of roselle callus mediated by Agrobacterium tumefaciens

Study on asymbiotic germination of Cattleya hybrid seed and development of protocorms during germinating

Technology of accelerating germination and desiccation tolerance in Archontophoenix alexandrae seeds

Characteristics of chlorophyll fluorescence and comparison of drought resistance among different species of karst plants

Seasonal photosynthetic responses of Phyllostachys edulis to environmental factors

Comparison of caloric values and ash contents in three palm species

Effects of light on maca callus growth, shoot induction and its survival rate

Rapid propagation of Dracaena cambodiana in vitro

Shoot regeneration from cotyledon of orange-heading Chinese Cabbage

Organic nutrition and the shoot growing trait of heavily retractive pruned longan trees

Cryopreservation of recalcitrant seed germplasm

Metabolomics approaches and their application in Botany

Comparative analysis on effect of Fe2+,Cu2+and Zn2+ to physiological characteristics of plant

Study on changes in concentrations of four endogenous hormones in different fruit positions during citrus fruit growth and development

Rule of growth and development on flower-bud of Mangifera indica cv.Zihuamang

Studies on the development of the leaf of Pinus kwangtungensis

Sex identification of Siraidia grosvenorii by isozyme analysis

Changes on leaf oxalate content in buckwheat growing under different nitrogen forms

Establishment of genetic transformation system of Populus euphratica and optimization of antibiotic concentration

Inducible formation of 2-(2-phenylethyl)chromones in cell suspension culture of Aquilaria sinensis

Preliminary study on the effect of low pH on the nodulation of Medicago sativa and corresponded mechanisms

Relation between endogenous hormones in bud and on-year or off-year fruiting of Ginkgo biloba

Effects of calcium signal system on anti-oxidant enzymes and membrane lipid peroxidation of citrus under low temperature

Effects of ethephon sprayed at early tillering stage on the activities of peroxidase, IAA oxidase and acid invertase in sugarcane in correlation to tillering

Preservation in vitro of medicinal plant Salvia prionitis

In vitro culture of different explants from Pogostemon cablin

Cloning and idientifcation of β-glucosidase gene of uncultured microorganisms from biogas digester

Studies on the kinetics of cell growth and nutrient consumption in cell suspension cultures of Corydalis saxicola

Activity assay of grain starch synthesis key enzymes in wild rice species of Yunnan

Physiological compare Merremia boisiana with Pueraria lobata under drought stress and rewatering conditions

Study on inducion and multiplication culture of bulblet from some parts of flower in Lilium

The cutting propogation of palmleaf raspberry(Rubus chingii)

Study on tissue culture of Dysosma versipellis

Tissue culture and rapid propagation of Lonicera confuse

Technology procedure of Tissue culture and Propagation of Siraitia grosvenorii

Advances in biotechnological research on Litchi(Litchi chinensis)

Effects of Cd on physiology and biochemistry and leaf ultrastructure of Seleng wormwood

Influence of the obligate parasite Cuscuta campestris on the leaf stomatal behavior of its host Mikania micrantha

Anthocyanin essence of the purple pigment and positive correlation of the anthocyanin content and the total ginsenoside content of the root tuber of Panax notoginseng

Preliminary study on the cell suspension culture of Eucommia ulmoides and secondary metabolite-chlorogenic acid

Effects of long-lasting brassinosteroid TS303 and propyl dihydrojasmonate on enhancing peanut resistance to chilling

Genetic analysis of meiotic behavior difference in plants from the same population of Zephyranthes grandiflora

Tissue culture of medical plant Erigeron breviscapus

Analysis of the chromosomal location of 45S rDNA in plants by CPD staining together with FISH

Expression of AtPSK2 gene in roselle cells

Allelopathic effects of rock plant Pogonatherum paniceum on crops

First report Emilia sonchifolia wilt caused by Ralstonia solanacearum

Allelopathic effects of Parthenium hysterophorun aqueous extract on germination of Bidens pilosa and Delavaya toxocarpa

Study of latex exploitation using depolymerizing compounds of microfilaments in rubber trees

Growth and biomass allocation of Artemisia annua from different seed sources

Effects of different treatments on cuttage cultivation and growth of Keteleeria calcarea

Peroxidase isozyme in sprouting seeds of genus Fagopyrum

Physiological and biochemical characteristics of Myricaria laxiflora,an endangered species in the Three Gorges Reservoir Area

Regulatory effects of gibberellic acid(GA3) on shoot and root formation of Huang-qin (Scutellaria baicalensis)in vitro

Inhibitory mechanism of extracts from Ginkgo biolaba sarcotesta on mushroom tyrosinase

Analysis of some quality characteristics in soybean mutants

Cloning and analysis of hmgr gene conserved fragments in Descurainia sophia

Purification and kinetic characteristics of an acid phos- phatase isoform from germinating pigeonpea seed

Existent states of anthocyanins in vacuole and their coloration effects in higher plants

A preliminary study on the water physiology of four plant species in karst area of Southwest China

Influences of different gibberellin and media on growth of seven cultivars of colour callalily

Isolation of protoplast from the Astragalus membranaceus

Electrophysiological mechanism of root length growth of Arabidopsis thaliana inhibition by JA-Me

Physiological mechanism in bitter melon infected with powdery mildew fungus

Changes of superoxide dismutase and estrase in processes of embryogenic calli induction of Eustoma grandiflorum

Effect of different wavelength lights on photosynthesis and growth of Siraitia grosvenorii

Endophytic fungi screening from Atracty lancea and inoculating into the host plantlet

Studies on several physiological and biochemical indices of the heat-resistance of Lilium longiflorum and its comprehensive evaluation

Glands structure of liquorice and its regulation mechanism on osmosis stress

Effect of Cadmium on physiological characteristics of mangrove Avicennia marina and Aegiceras corniculatum seedlings

Effect of aluminium on protection enzyme system and proline of Brugiera sexangula seedlings

Analysis on the different characteristic of calcium in the seed of Cycas guizhouensis

Advances on screening of Cd-hyperaccumulators and transgenic improvement of plant in Cd-accumulation

Construction of a plant expression vector containing antifreeze protein(afp)gene and it's transformation into banana(Musa AAA Cavendish) embryogenic cell suspension

Construction and transformation of antisense RNA expression vector of Arabidopsis phytochrome A gene

Studies on pollen germination and storage of Cycas species

Study on in vitro selection of Pogostemon cablin resistant to bacterial wilt

Screening a group of cellulose-degradation bacteria and their zymogenic characteristics

Effects of Cr⁶⁺on the growth and some physiological characters of tobacco by tissue culture

Comparative analysis on effect of Fe²⁺,Cu²⁺and Zn²⁺ to physiological characteristics of plant

Regulatory effects of gibberellic acid(GA₃) on shoot and root formation of Huang-qin (Scutellaria baicalensis)in vitro

Changes of nuclear ultrastructure,nDNA and antioxidant enzymes in leaves of Lemna minor induced by Cd²⁺

Effect of exogenous NO on the growth and inorganic ion content of Festuca arundinacea seedlings under Pb²⁺ stress