2 population reached 61.04% and 69.60% in two floral traits,respectively,and the polygenes heritabilities were zero. Thus the male floral traits were quantitative inheritance,and the heritabilities were relatively high. In breeding practices,it could thus be seen that,gentic selection of floral charactors in cucumber should be performed at earlier separate generations.]]> 2015/12/14 13:25:13 1, SUI Yi-Hu¹, JIAN Xing²]]> MIAO Yong-Mei¹, SUI Yi-Hu¹, JIAN Xing² 37true Lolium perenne)cultivar ‘Top Hat 2' was used as explants. Firstly, this paper focused on the effect of four ways of explants treatments(embryo end half a caryopsis, slitting embryo end half a caryopsis, small pieces of embryo end for caryopsis, small pieces of caryopsis)and two ways of surface sterilization methods(disinfection with sodium hypochlorite solution, disinfection with mercuric chloride)on callus induction; Secondly, we studied the effects of different plant growth regulators(2, 4-D and 6-BA)and their concentrations on callus induction by randomized block design of two factors; Thirdly, we studied the effects of different plant growth regulators(NAA, 6-BA and ZT)and their concentrations on adventitious bud differentiation by orthogonal test of three factors and four levels; At last, we separately studied the effects of 0.10 mg·L^-1NAA and IBA on adventitious root induction. The results were as follows: Under the same conditions, explants of full end half a caryopsis embryos were surface sterilized with 75% alcohol for 1 min, then treated with sodium hypochlorite solution 1 mL·L^-1 Tween 20(the chlorine content>10%)for 30 min, the rate of callus induction were better than the other's; The explants inoculated on MS medium with 5 mg·L^-1 2,4-D and 0.03 mg·L^-1 6-BA(pH=5.8), provided us a relatively high induction rate of 68.7% and high quality callus; The most high differentiation rate was 56% when adventitious buds were inoculated in MS with 0.5 mg·L^-1NAA, 0.5 mg·L^-1 6-BA and 0.9 mg·L^-1 ZT. The best rooting medium was 1/2MS+IBA with nothing plant growth regulator obtaining stout root, then the regeneration rate was 90%. This paper has established efficient tissue culture regeneration system for perennial ryegrass and laid a good foundation for conducting highly efficient genetic engineering.]]> 2015/12/14 13:25:13 *]]> WEI Shu-Qiang, QIAN Yong-Qiang, LIU Jun-Xiang, SUN Zhen-Yuan^* 36true Pogostemon cablin syn. P. patchouli,is a perennial bushy herb introduced from Southeast Asia and has been extensively cultivated in South China,for both utilization in pharmaceutical and perfume industry. Patchouli cultivated in different regions has diverse morphological characteristics and essential oil constituents,which infect its therapeutic properties directly. It is necessary to established a cultivar with high quality of essential oil. This study aims to establish an efficient system of plant regeneration from the leaf-disc of Pogostemon cablin. The leaves of in vitro patchouli seedlings were used as materials. The effects of donor plant's age,leaf position and the concentration of plant growth substances used single or combined on plant regeneration of P. cablin were studied. The results showed that low concentration's benzylaminopurine(BA)combined with naphthaleneacetic acid(NAA)induced the high frequency of prolific adventitious shoots regeneration directly from leaf with less or without callus,the age of donor plant and position of leaf on stem both effected the frequency of shoot regeneration and the mean number of shoots. The high efficient plant regeneration system was established from leaf disc explants prepared from the leaves of the second nodes of thirty-day old-in vitro plant. The culture medium for the prolific shoot inducing was MS culture medium supplemented with 0.1 mg·L^-1 NAA and 0.5 mg·L^-1 BA. The regeneration frequency was 100% and the maximum mean number of shoot per leaf disc was 96.5. Regenerated shoots were elongated and rooted on the half strength MS medium with 0.5 mg·L^-1GA₃ and 0.1 mg·L^-1 NAA. The rooting frequency was 100%. The plantlets with well-developed roots were successfully acclimatized in the greenhouse,and planted in the field with a survival rate of 96%. The established plant regeration system from leaf-disc of P. cablin would lay foundation for the gene transformation of P. cablin and its fast breeding for good cultivars.]]> 2015/12/14 13:25:14 1*, OUYANG Pu-Yue², ZENG Qing-Qian¹, HUANG Shan-Shan¹]]> MO Xiao-Lu^1*, OUYANG Pu-Yue², ZENG Qing-Qian¹, HUANG Shan-Shan¹ 35true Pinus kesiya var. langbianensis,and to study the role of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase in regulation of resin biosynthesis,the transcriptome of bark of Pinus kesiya var. langbianensis was sequenced by Next-Generation Sequencing. First,a fragment of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase gene was obtained from Pinus kesiya var. langbianensis transcriptome after gene assemble and gene function annotation. The special primers were designed according to the fragment of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase. RNA of injured bark was extracted by Trizol method. The full length gene of PkHDR was cloned from Pinus kesiya var. langbianensis by Reverse Transcription-Polymerase Chain Reaction(RT-PCR)and rapid-amplification of cDNA ends(RACE). Bioinformation analysis showed that the obtained full cDNA sequence of PkHDR had 1 876 bp. It was consisted of 1 464 bp open reading frame(ORF)which encoded 487 amino acid. Homology analysis indicated that the deduced PkHDR protein shared 99% identities with the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase came from Pinus densiflora. Subcellular localization and structural domain analysis showed that the transit peptide sequence(A1-A61)and multiple conserved functional sites(A143,A234,A288,A371)of plant HDR protein were found in the deduced coding sequence of PKHDR. Phylogenetic analysis revealed that the evolutionary relationship of PkHDR protein was the closest to Pinus densiflora HDR protein. Reverse transcription polymerase chain reaction(RT-PCR)detection showed that PkHDR gene expression was up-regulated by wounding treatment. The full cDNA of PkHDR from Pinus kesiya var. langbianensis was cloned and the reverse transcription polymerase chain reaction(RT-PCR)showed that PkHDR was involved in regulation of resin biosynthesis in Pinus kesiya var. langbianensis. These results would provide important information to reveal the resin biosynthesis in Pinus kesiya var. langbianensis. And this study also can be applied in the research of the high yield of resin variety molecular breeding.]]> 2015/12/14 13:25:14 1,2, ZHOU Xu³, BI Wei^1,2, YANG Yu-Ming², LI Jiang², WANG Juan^2*]]> WANG Yi^1,2, ZHOU Xu³, BI Wei^1,2, YANG Yu-Ming², LI Jiang², WANG Juan^2* 34true Ft4CL)from Fagopyrum tatarium. Using buckwheat species ‘Xi Qiao No.2', according to the conserved squences of 4CL from GenBank,a pair of degenerate primer was designed and synthesized. Through RT-PCR(reverse transcription PCR)technique,the conserved fragment of Ft4CL was amplified from the total RNA of F. tataricum flower buds. Then,the RACE technique(rapid-amplification of cDNA ends)was performed,and the 5' end and 3' end of Ft4CL were succesfully amplified,respectively. The complete cDNA of Ft4CL was obtained by splicing the above sequences,and a pair of gene-specific premie was synthesized to amplify the ORF(open reading frame)regine of Ft4CL. Using DNAman software to deduce the ORF sequence of Ft4CL to the amino acid sequence,its homologous with other 4CLs were analyzed by NCBI Blast tool. The secondary structure of Ft4CL was predicted by SOPMA(http://pbil.ibcp.fr),multiple sequence alignment was performed by DNAman software, and phylogenetic tree was built with neighbor-joining method by MEGA 5.0. The results were as follows: the similarity of Ft4CL with F. esculentum(HM149785)showed the hightest level(up to 94%)and it ranging form 66%-75% with other plant 4CLs. Multiple sequence alignment results showed that the Ft4CL had conserved motifs of BOXⅠand BOXⅡ near by the C-terminus,but it had relatively low similarity with other 4CLs at the C-terminus. According to the phylogenetic tree analysis results,the selected 4CLs were grouped into 2 cluster,Ft4CL,Arabidopsis thaliana 4CL1,and A. thaliana At4CL2 belonged to ClusterⅠ. In conclusion, the results could provide basic data for in-depth study of Fagophrum tatarium flavonoid pathway. Furthermore,this study indicated that Ft4CL could be a new candidate target gene for developing high flavoniod F. tatarium by metabolic engineering technology in future.]]> 2015/12/14 13:25:15 1, GAO Fei², WANG An-Hu³, LI Cheng-Lei², CHEN Hui², WU Qi^2*]]> LING Yao¹, GAO Fei², WANG An-Hu³, LI Cheng-Lei², CHEN Hui², WU Qi^2* 33true Dioscorea bulbifera microtubers as material,the anatomical structure and the physiological and biochemical indexes of microtubers during low-temperature conservation were observed and measured,the genetic stability of germination seedlings after low-temperature conservation was also tested in this article. The results were as follows: HE staining method was complicated,whose effect was difficult to observe. Compared with HE staining method,safranin fast green staining was much simpler,whose effect was better. Therefore,Safranin-fast green method was more suitable for staining of D. bulbifera microtubers; Without low temperature conservation,the starch grains in the cells around D. bulbifera microtuber has not begun to be exploited and be still conserved. When D. bulbifera microtuber treated with low temperature,the starch grains in the cells surrounding microtuber began to disappear. The longer the conservation time is,the more the cell number with no starch grains also increased. The starch grains in cells had started to translate into other substances for cell life activities during low temperature storage. When conserving for 90 d,the cell whose starch grains disappeared extended to the middle of the microtuber; During the low temperature conservation period,the antioxidase,the amylase activity,the proline content and the soluble sugar content of D. bulbifera microtubers all showed an increasing trend; During the low temperature conservation of D. bulbifera microtubers for 0-90 d,SOD activity during 18-54 d continued to increase,and SOD activity during 54-90 d still remained unchanged; POD activity increased significantly within 18-36 d,maintained stability within 3-54 d,increased significantly during 54-72 d,kept stable again in 72-90 d; CAT activity trends is consistent with the SOD activity,which continued to increase in 18-54 d,and remain unchanged in 54-90 d; α-amylase and total amylase activity increased rapidly in 18-36 d,had no significant change in 36-54 d and continuously increased significantl in 54-90 d; β-amylase activity increased significantly in 18-54 d,maintained stability within the 54-72 d,and began to increase significantly within the 72- 90 d; Soluble sugar content increased significantly within 18-36 d,had no significant change within 36-54 d,and began to increase significantly during 54-90 d; Proline content had no change in 18-36 d,increased significantly in 36-72 d,and remained unchanged in 72-90 d. There was no significant difference between the seedling germinated from D. bulbifera microtubers stored at low temperature for 90 d and the seedling germinated from D. bulbifera microtubers without low temperature conservation in morphology(average plantlet length,average leaf number,average interstem length and average root number),DNA content,physiological index(total chlorophyll content,superoxidase dismutase activity,catalase activity,peroxidase activity,soluble sugar content and soluble protein content)and its leaf photosynthetic characteristics parameters(net photosynthetic rate,stomatal conductance,intercellular CO₂ concentration,transpiration rate,stomatal limitation value,water use efficiency and instantaneous carboxylation rate)and chlorophyll fluorescence parameters(initial fluorescence,maximal fluorescence,the most photochemical efficiency of PS II,the potential photochemical efficiency of PS II,the actual photochemical efficiency of PS II,captured excitation energy efficiency of open PSII reaction center,photochemical fluorescence quenching coefficient and non-photochemical fluorescence quenching coefficient),which indicated that low temperature conservation of D. bulbifera microtuber could not cause genetic variation of the seedling.]]> 2015/12/14 13:25:15 *, XIA Jin-Hua, LIN Guo-Wei, WANG Ai-Bin, KE Wei-Zhong]]> YIN Ming-Hua, HONG Sen-Rong^*, XIA Jin-Hua, LIN Guo-Wei, WANG Ai-Bin, KE Wei-Zhong 32true GrWRKY5 was cloned from the spire of Gentiana rigescens. Its bioinformatic analysis was performed to predict its functions and its plant overexpression vector was also constructed. The gene specific primers were designed according to the transcript sequence of GrWRKY5 from the transcriptome of triennial G. rigescens. The open reading frame(ORF)of GrWRKY5 was obtained by Reverse Transcription-Polymerase Chain Reaction(RT-PCR). Then TA cloning,sequencing,and sequence analysis were performed. Entry vector pENTR^TM2B-GrWRKY5 was constructed,and the plant overexpression vector of pK₂-35S-GrWRKY5 was obtained after LR reaction. The ORF of GrWRKY5 has a length of 591 bp and encodes a predicated protein of 196 amino acids. The GenBank accession number for GrWRKY5 is KF922375. The tryptophan(W),arginine(R),lysine(K),tyrosine(Y)between 167 and 170 in WRKY protein consist of the "WRKY" symbolic structure. Sequence analysis showed that GrWRKY5 was a member of “WRKY” superfamily. Results of bioinformation software on line analysis showed that the theoretical isoelectric point(pI),the aliphatic index and the instability index of GrWRKY5 protein were 6.29,61.37 and 57.80,separately. The GRAVY(grand average of hydropathicity)of GrWRKY5 protein was -0.708,which indicated that it belonged to hydrophilic protein. GrWRKY5 protein contained more than 20 kinds of amino acids,and the aspartic acid(Asp)and proline(Pro)accounted for the highest content(8.1%),while Cysteine(Cys)content was the lowest accounting for only 1.0%. Results of phylogenetic analysis showed that GrWRKY5 was at the same evolutionary branch with AtWRKY27,a member of class Ⅱe,and that GrWRKY5 was close to the CrWRKY22 protein of Catharanthus roseus and the VvWRKY22 protein of Vitis vinifera and was far from the JcWRKY47 protein in Jatropha curcas and the TcWRKY27 protein in Theobroma cacao. Results of BLASTp showed that GrWRKY5 protein had the highest identity(69%)with the BnWRKY27-1 protein in Brassica napus,while it had the lowest identity(31%)with the AaWRKY22 protein in Arabidopsis thaliana. The plant overexpression vector pK₂-35S-GrWRKY5 which was suitable for agrobacterium tumefaciens and gene gun-mediated transformation was successfully constructed base on entry vector pENTR2B and destination vector pK₂GW7 of Gateway system. The successful construction of plant overexprossion vector pK₂-35S-GrWRKY5 will provide a foundation for its genetic transformation into Arabidopsis thaliana and Gentiana rigescens and provide a basis for the further study of gene function.]]> 2015/12/14 13:25:16 1,4, LI Fu-Sheng¹, LI Tao², LI Cai-Xia², ZHANG Xiao-Dong^2*, WANG Yuan-Zhong³]]> WANG Cai-Yun^1,4, LI Fu-Sheng¹, LI Tao², LI Cai-Xia², ZHANG Xiao-Dong^2*, WANG Yuan-Zhong³ 31true Senecio scandens for probing its sequence features and composition, and gene's function by applying a series of bioinformatics software. The structure properties of primary, secondary, domain and tertiary of the amino acid sequence were analyzed, too. And the phylogenetic relationships among α-Importin of S. scandens with other species were also constructed. These exploration found that this gene encoded a length of 529 amino acid polypeptide with the calculated molecular weight of 58.46 kDa, and its theoretical isoelectric point was 5.08. It shared 84.0% identity with tobacco α-Importin gene(GenBank ID: ABM05487.1)by amino acid sequence alignment. The structure analysis showed its secondary structure was composed of α helix protein, a random coil and an extended main-chain. The function domain probing found the target gene was composed domains of IBB and ARM. While its three-dimensional structure was mainly consisted of four structural domains. In conclusion, this study found that the function of this α-Importin gene was regulation of hormone secretion, a signal transducer, growth factor, transcription and transcription regulation with higher rank of probability ratio. The current study also founded that this α-Importin gene could play important roles in many biology processes such as non-apoptotic programmed cell death, defence response of higher plants against pathogens, hormone receptor effect and gene transcription reactions and expression. According to the exploring of the properties of gene sequences, structure and function of α-Importin in S. scandens, this paper would provide a reference for similar analysis of the relationship between the structure and function of α-Importin genes in other species.]]> 2015/12/14 13:25:16 1, XU De-Lin¹, ZHANG Lin¹, QIAO Xiao-Ying², LUO Shao-Yuan², QIAN Gang^1*]]> CHU Shi-Run¹, XU De-Lin¹, ZHANG Lin¹, QIAO Xiao-Ying², LUO Shao-Yuan², QIAN Gang^1* 30true Gentiana macrophylla. These compounds have widely biological and pharmacological effects, such as stomachic, choleretic, anti-hepatotoxic activities, anti-inflammatory, antifungal and antihistamine activities. Secoiridoids belonged to monoterpenoid, were biosynthesized via the secoiridoid pathway(sometimes also called “ridoid pathway”)in high plant. 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(HDR)is one of the key enzymes in the pathway of iridoid biosynthesis. In this paper, we cloned the gene sequence of HDR from G. macrophylla, and analyzed the characteristic of sequence and expression patterns in order to know its roles in secoiridoid biosynthesis. Based on our library generated by high-through sequencing of G. macrophylla transcriptome, we cloned HDR gene from G. macrophylla(named as GmHDR)by RT-PCR. And the GmHDR coding amino acid sequence characterization, such as physicochemical characteristics, signal peptide, transit peptide, subcellular localization, conserved domain and secondary structure, analyzed with bioinformatics methods. Then we also detected the expression patterns of GmHDR in different parts of G. macrophylla by real time PCR. One 1 630-bp length sequence of GmHDR gene was obtained from G. macrophylla. GmHDR contains a completed open reading frame(ORF)of 1 392 bp, which encoded a polypeptide with 463 amino acids. GmHDR, the encoding protein by GmHDR, has high homology(identities ≥ 84%)to HDR proteins from Rauvolfia verticillata, Tanacetum parthenium and other plants. One neighbor joining tree was constructed to show evolution ship between GmHDR and HDR proteins from other plants using MEGA5.2 soft. The phylogenetic tree also gave a same conclusion that GmHDR had a closed relation with HDR proteins from Catharanthus roseus and Rauvolfia verticillata. Further analysis with bioinformatics methods showed that GmHDR was one protein without transmembrane domain,and had one transit peptide with 36 amino acids predicted with ChloroP server. These indicated that GmHDR protein might be located in the chloroplast. Real time quantitative PCR results showed that GmHDR had a very high expression level in the flowers of G. macrophylla, and GmHDR gene expressed lower in roots, stems and leaves of Gentiana macrophylla. Conclusion: The sequence of GmHDR had a lot of similar features with HDR proteins from other plants, such as conserved four-cysteine sites, transit peptide in their N terminal. GmHDR mainly expressed in the flowers of G. macrophylla. These results indicated that GmHDR may be principal involved in terpenoid biosynthesis, which are accumulated in flowers of G. macrophylla. The research is not only very helpful for research on HDR roles in G. macrophylla,but also will lay the foundation for the further study on biosynthetic pathway of secoiridoid compounds in G. macrophylla.]]> 2015/12/14 13:25:17 1, KONG Wei-Wei1, ZHENG Peng1, HUA Wen-Ping1,2*]]> CEN Wen1, KONG Wei-Wei1, ZHENG Peng1, HUA Wen-Ping1,2* 29true Lysimachia christinae Hance(Christina loosestrife),a mat-forming perennial herb from the Primulaceae family, is a famous Chinese medicinal material and excellent ground cover plant for landscaping. The natural triploid of L. christinae is reported here so as to provide new reference for the development and utilization of L. christinae. In this study, L. christinae plants introduced from Lion Mountain population in Wuhan and Moon Mountain population in Huangshi were cultivated in the same text site. And then the plant morphology and the stoma size were observed. The chromosome numbers and karyotypes were investigated by the conventional pressing plate method. The results showed that the leaf length, leaf width, leaf area, length of petal, petal width, petal area and leaf stoma area of L. christinae from Moon Mountain population in Huangshi were significantly higher than those of L. christinae from Lion Mountain population in Wuhan. In addition,the karyotype formula of L. christinae from Moon Mountain population in Huangshi was 3n=36=3m+3sm+6st+24t,which was different with that of L. christinae from Lion Mountain population in Wuhan(2n=24=2m+2sm+4st+16t). Therefore, in this paper, the L. christinae from Moon Mountain population in Huangshi was identified as natural triploid based on abovementioned morphology and chromosome datum. The discovery of natural triploid in L. christinae enriched the polyploidy data of L. christinae. It would not only provide reference for research on genetic diversity and evolution of L. christinae,but also supply the excellent material for medicine, landscaping and breeding.]]> 2015/12/14 17:02:59 1, CHEN Long-Qing², ZHENG Wei^1*]]> LIU Xiao-Ming¹, CHEN Long-Qing², ZHENG Wei^1* 28true Roegneria grandis(2n=4x=28)and to provide cytological evidence for the taxonomic treatment of R. grandis, artificial distant hybridization between R. grandis and Elymus wawawaiensis(2n=4x=28)was carried out. Three F₁ hybrid plants were obtained successfully. The general morphological appearance of the intergeneric F₁ hybrid was intermediate between the Roegneria grandis and Elymus wawawaiensis. For estimation of pollen fertility study, pollen grains of hybrids were examined after staining in I₂-IK solution. Pollen grains of parental species were highly fertile. The percentage of stained pollen grains of Roegeria grandis reached 94.6%, and 90.5% pollen grains of Elymus wawawaiensis were fertile. Pollen grains of the hybrid between Roegeria grandis and Elymus wawawaiensis were sterile. Chromosome pairing at meiosis metaphase I(MI)of Roegeria grandis and Elymus wawawaiensis and their F₁ hydrid were analyzed. Meiosis of the parental tetraploid species was quite regular with 14 bivalents. Most of them were ring bivalents. A low frequency of univalent(0.04/cell)was observed in Roegeria grandis. Chromosome pairing at meiosis metaphase I(MI)was observed in 83 MI cells in the F₁ hybrid. Meiotic pairing in the F₁ hybrid averaged 6.46 bivalents per cell, varying from 5 to 8. 7 bivalents occurred in 35.2% of the MI cells, 42.59% of the MI cells had 6 bivalents, and 8 bivalents were observed in several MI cells. All the metaphase I cells contained unpaired chromosomes,ranging from 10 to 18,with a mean of 14.66 univalents per cell. 0.14 trivalents per cell were observed. The meiotic configurations of the F₁ hybrid were 14. 66 I+6.46 II+0.14 III. The chiasmata per cell were 9.83 with C value of 0.35. The results were as follows:(1)Roegeria grandis and Elymus wawawaiensis shared a homologous St genome. The other genome of Roegeria grandis was not St or H genome. The genome constitution of R. grandis was not StSt^g genome;(2)A low frequency of trivalents(averaged 0.14 per cell)were observed in the F₁ hydrid. They can probably be attributed to structural rearrangements between chromosomes of R. grandis and Elymus wawawaiensis, or a certain degree of homoeology between the St and Y genomes in these species. Further study needs to be done to investigate the oringin of Y genome;(3)The homology of St genome of Elymus species from different geographical origins with the St genome of Roegeria grandis was different. Elymus lanceolatus and E. wawawaiensis were from north America, and E. sibiricus and E. caninus were from Asia. The homology of St genome of Roegeria grandis with the St genome of Elymus species from north America was higer than that with the St genome of Elymus species from Asia. The reason remained to be discussed in the future.]]> 2015/12/14 17:03:00 *]]> YANG Cai-Rong, ZHANG Hai-Qin, SHA Li-Na, ZHOU Yong-Hong^* 27true CsTS genes function studies of tea. Two gene sequences logged on NCBI were cloned,the expression of enzymes activities was verificated through HPLC and LC-MS and phylogenetic analysis was implemented within the searching result. The expression of enzymes in E. coli suggested CsTS and CsGS had different catalytic activities although their high homology. The primary and secondary protein structures of CsTS and CsGS were analyzed and the result showed that there was no significant difference between them, but the third protein structure analysis by homology modeling in the catalytic sites showed three differences in the catalytic sites, which could lead to the difference of enzyme activity. In this research, CsTS gene sequences were firstly identified as the members of GS gene family through phylogenetic analysis. According to cellular localization prediction, CsTS should belong to cytoplasmic GS(GS1), for its kinship close to grape, upland cotton, rubber tree, arabidopsis etc.]]> 2015/12/14 17:03:01 *]]> CHEN Qi, JIANG Xue-Mei, MENG Xiang-Yu, ZHANG Zheng-Zhu, WAN Xiao-Chun^* 26true Ps-mnp1 from Pleurotus sapidus PS1 contributing a lot to the lignin biodegradation activity is important for the analysis of protein structure and functions of MnP and the understanding of manganese peroxidase gene function and transcriptional regulation of Pleurotus sapidus. Based on the analysis of ribosomal rDNA Intenal Transcribed Spacer(ITS)sequences,the classification status of the strain PS1 was defined. Ps-mnp1 was cloned by using the methods of Genome Walking-PCR and Reverse transcription-PCR with primers designed from some white-rot fungi which contains the conserved sequences of the known manganese peroxidase genes(GenBank No. KP189358.1). We used a variety of modern bioinformatics technology softwares to clone the manganese peroxidase gene and analyze the protein structure of Ps-mnp1. For example, after the cloning full length DNA sequence of genome walking method, prediction of the full cDNA sequence of manganese peroxidase gene by using contigexpress splicing software; comparison the DNA and cDNA nucleotide sequences alignment analysis by using BioEdit software; prediction the RNA splice site through Augustus website,following the comparison and correction of Ps-mnp1 sequences by using the the NCBI database; understanding the compositions of the intron and exon by using online software Gene Structure Display Server contrast of white rot fungus manganese peroxidase gene. The results obtained in the sequence analysis of Ps-mnp1 from Pleurotus sapidus indicated that the full length of the DNA was 1 854 bp containing 14 exons and 13 introns. Meanwhile, exons and introns composition analysis of the genes Ps-mnp1 from Pleurotus sapidus, compared to other white-rot fungi manganese peroxidase genes, suggested that P. ostreatus and P. sapidus belonged to the same genus Pleurotus, but the manganese peroxidase gene structures between them were entirely different. The Ps-mnp1 gene included a signal peptide of 20 amino acids and held Open Reading Frame of 1 095 bp,with start codon ATG and stop codon TAA,encoding 364 amino acids. The results of protein phylogenetic analysis by MEGA 5.1 software revealed that Ps-mnp1 and 6 strains of white rot fungus protein clustered on short branches MnPs,and differentiated 5 long branches of MnPs group formed by disulphide bond. In addition,the 3D structures of Ps-mnp1 were built using homology modelling technique, and the results showed that the binding sites of the protein ligand included 1 Fe heme,2 Ca ions and 1 Mn(II),and the those sites were non-conservation. The results also suggested that the modelling similarity was 72.51% compared with P. eryngii VPs. This paper could establish the basis of the study in the structure and function of the Ps-nmp1. It is further helpful to offer reference on the variation of protein engineering of the Ps-mnp1.]]> 2015/12/14 17:03:01 1*, YANG Chun-Cheng1, ZHU Yu1, LUN Zhi-Ming2, ZHANG Jing2]]> YIN Li-Wei1*, YANG Chun-Cheng1, ZHU Yu1, LUN Zhi-Ming2, ZHANG Jing2 25true StWRKY2 has not been reported. Plant WRKY proteins are found as an important class of transcription factors in recent years, which are widely involved in biotic stress, abiotic stress and growth development regulation, and they are mainly divided into three groups. WRKY2 was cloned from potato by homology cloning, with length of the coding region 1 065 bp,encoding 355 amino acids by DNA sequencing. With the amino acid sequence of StWRKY2 blasting in NCBI, 19 homology amino acid sequences were selected to analysis conservative domain. Amino acid analysis showed that the StWRKY2 contained 1 conserved WRKY domain(TTGACC)and belonged to the second group of WRKY family members, and its zinc-finger structure was C-X₅-C-X₂₃H-X-H. Phylogeny tree results showed StWRKY2 was the most closest to SlWRKY7(Solanum lycopersicum)with 96% similarity, and the similarity of a WRKY protein of tobacco was 86%. The protein-GST(glutathione-S-transferase)complex was obtained in Escherichia coli by using the method of prokaryotic expression. StWRKY2-GST could combine W-box in vivo by Electrophoretic Mobility Shift Assay analysis,but only GST protein could not combine W-box. StWRKY2-GST combination with W-box could be competed by cold probe(unlabeled probe). And the experiment results also showed that StWRKY2-GST could not combine mutation W-box,this suggested StWRKY2-GST combined W-box with specificity. Analysis of StWRKY2 expression level in the root, stem and leaf tissue through the real-time fluorescence quantitative PCR,the results showed that the gene was mainly expressed in the root. Next was leaf and stem. In order to further study the possible function of the gene,potato seeding from tissue culture were treated under 10 μmol/L low phosphorus,10 μmol/L low potassium,200 mmol/L NaCl,400 mmol/L PEG solution and 4 ℃ low-temperature treatment for 6 h,real-time fluorescence quantitative PCR showed that this gene expression level decreased significantly after low phosphorus treatment, expression of StWRKY2 increased significantly after 200 mmol/L NaCl and 400 mmol/L treatment. Expression level of StWRKY2 had no obvious change after low potassium and 4 ℃ low temperature treatment. The results indicated that StWRKY2 could in response to low phosphorus,NaCl and PEG three kinds of abiotic stresses. These results would lay a solid foundation for further study of potato WRKY2 gene function.]]> 2015/12/14 17:03:02 * ( College of Agronomy, Sichuan Agricultural University, Chegndu 611130, China )]]> LI Li-Qin, WANG Xi-Yao^* ( College of Agronomy, Sichuan Agricultural University, Chegndu 611130, China ) 24true Arabidopsis,PKS5(protein kinase SOS2-like 5), a serine-threonine kinase, involves in the response to the external high pH stress based on the study of its loss-of-function mutant. Whereas, the fine functions of the domains resided in PKS5 are not currently well determined. We report here the dissection of domains of PKS5 in the activity of phosphorylation against MBP(myelin basic protein)and AHA2(one of the Arabidopsis thaliana isoform of PM H⁺-ATPases), which is the specific substrate of PKS5 in vivo, using the assay of phosphorlation in vitro via expressing the distinct PKS5 mutant versions in bacteria using the PKS5 cloning from plants employing PCR approach. The results showed that the point mutated PKS5-2 lost its activity, PKS5-4, PKS5-5 and PKS5-9 displayed no difference in autophosphorylation and the MBP phosphorylation. Moreover,autophosphorylation and the AHA2 phosphorylation of the point mutated PKS5-6 and PKS5-7 increased compared with PKS5 and the PKS5-7 activity was higher than PKS5-6. Taken together, the specific point mutations in PKS5 altered its activity of autophosphorylation and the substrate phosphoylation of BMP and AHA2. The effects due to the alteration of mutations resided in PKS5 on the activity of phosphorylation were comparable within the various PKS5 mutated protein versions. The results of this study would provide the basis for pinpointing the functional domains of PKS5 and the mechanism of functions of PKS5 in planta.]]> 2015/12/14 17:03:03 1,2, JIAO Cheng-Jin¹, CHEN Quan¹, MA Wei-Chao¹, AN Jian-Ping¹, HU Li-Ping^{1, 2}]]> ZHAO Fei-Yi^1,2, JIAO Cheng-Jin¹, CHEN Quan¹, MA Wei-Chao¹, AN Jian-Ping¹, HU Li-Ping^{1, 2} 23true 2015/12/14 17:03:03 *, WANG Jin, YANG Shu-Ting, TU Xun Liang, LAI Jing]]> CHEN Zhong-Gang, LÜ Xiu-Lan^*, WANG Jin, YANG Shu-Ting, TU Xun Liang, LAI Jing 22true 8 conserved domain,the corresponding cDNA sequence(XM-004245875.1)was located on SL2.40 region in the chromosome 8, with genomic DNA length 4 089 bp, the alignment region in chromosome 8 between 57228847-57232935 bp, and composed of two exons and one intron. The tomato RGA4 protein was an unstable globular secretory protein without transmembrane structure. Tomato RGA4 protein was regarded as an orthologous of potato Rpi-blb1 protein because they were similar in secondary structure composition,gene sequence composition,gene mapping,etc,but their minor differences in gene evolution and composition could result in different functions. This study would provide theory for cloning late blight resistant gene and its utilization in tomato breeding of disease resistance.]]> 2015/12/14 17:03:04 *, RUAN Xian-Le, FAN Xiao-Fang, CHEN Long]]> WU An-Quan, WANG Jun-Sheng^*, RUAN Xian-Le, FAN Xiao-Fang, CHEN Long 21true Cerasus avium has been conducted in this study. Selfing set,hybridization affinity and fruit setting percentage were investigated and analysed. The results were listed as follows:(1)There was a high and low self-compatibility in C. pseudocerasus and C. avium,respectively;(2)Each hybridization combination showed high affinity after two weeks of pollination,with 57.24%-93.03% of hybrid fruit setting. Different levels of embryo degeneration were observed among hybrids after five weeks of pollination,especially in the distant hybridization;(3)The fruit setting rate between cultivars and wild germpalsms was significantly higher than between cultivars in interspecific hybridization combinations. While there was an opposite result in distant hybridization,as ‘Hongdeng' the female parent,the fruit setting rate when using cultivars as male parent was higher than as wild germplasms male;(4)Each cross-combination has different setting percentage influenced significantly by the male parent. We conducted a preliminary analysis and discussion on these results.]]> 2015/12/14 17:14:56 1, WANG Xiao-Rong^{1, 2*}, CHEN Tao¹, ZHANG Hong-Wei¹, DU Tao¹]]> DU Han-Mei¹, WANG Xiao-Rong^{1, 2*}, CHEN Tao¹, ZHANG Hong-Wei¹, DU Tao¹ 20true Brassica oleracea var. italica),to provide a new Ogu marker and reference for broccoli germplasm collection,utilization and molecular marker assisting selection, specific primers P1/P2 were designed to amplify twenty broccoli germplasm genomic DNA according to the conserved sequences of orf138 gene. A 392 bp specific fragment was detected in twelve infertile broccoli genotypes by PCR amplification,eight fertile genotypes with no band,harmonized with morphology in field. The sequencing and comparison showed the sequence(Genbank accession code:HQ149728)as high as 100% homologous to the reported orf138(Genbank accession code:Z18896.1)in radish. Sequence homology comparison detected orf138 gene contained some variation sites. In this study,the molecular identification of broccoli CMS,was helpful to further elucidate the mechanism of CMS abortion,to guide creation new CMS lines,and to provide a theoretical basis for heterosis efficient utilization.]]> 2015/12/14 17:14:57 1, PEI Xu-Li², TANG Zheng^1*, LIU Qing¹, ZHANG Xiao-Ling¹, LUO Tian-Kuan¹, ZHU Shi-Yang¹]]> JING Zan-Ge¹, PEI Xu-Li², TANG Zheng^1*, LIU Qing¹, ZHANG Xiao-Ling¹, LUO Tian-Kuan¹, ZHU Shi-Yang¹ 19true Salvia is the largest genus,comprising approximately 1 000 species. This genus is commonly found in central-South America,central Asia-Mediterranean,and East Asia. In East Asia,approximately 84 Salvia species are found in China. Many species of Salvia not only have economic values,but also have important visual values. Salvia has always been considered as monophyletic because its lever-like stamens. However,the tremendous diversity of morphological characteristics and vegetative habit of Salvia have led to long-existing and unresolved problem regarding infrageneric boundaries. Meanwhile,extensive in distribution and multiple variety and difference of morphology in Salvia species bring difficulty to classification and identification of Salvia. Besides,in the field survey and collection process,we found that it was very difficult to classification subg. salvia and subg. sclarea in nutrition stage. Epidermal hair were the most common appendages on plant leaves,the distribution characteristics and morphological characteristics important means of plant identification and taxonomic study. Micro morphological characters of 19 Salvia samples of 18 species and 1 forma,including type of leaf,shape of epidermis cell,type of anticlinal wall,type of stomata and type of epidermal hair were observed and measured using light microscope and environment scanning electron microscope,in order to provide foundation for taxonomic delimitation study of Salvia. The results showed that the characters of leaf epidermal and epidermal hair of these 19 samples under the environment scanning electron microscope display obvious diversity,the shapes of leaf epidermal cells were usually irregular or polygonal; the patterns of anticlinal walls were sinuous or sinuate,rarely straight or bow. The stomatal apparatuses could be classified into two types,anomocytic type and paracytic type. Anomocytic type was very common. According to their micromorphological characteristics,the epidermic hairs could be classified into 4 categories,peltate glandular hair,capitate glandular hair,short nonglandular hair and long nonglandular hair,respectively. These leaf epidermal morphological characters could provide evidence to identify and study the systematic relationships of taxa of Salvia.]]> 2015/12/14 17:14:57 1, LIU Shi-Yong¹, WANG Long^{1, 2}, WANG Hong-Yu¹, ZHANG Li^1*]]> WANG Tao¹, LIU Shi-Yong¹, WANG Long^{1, 2}, WANG Hong-Yu¹, ZHANG Li^1* 18true Camellia sinensis(L.)O. Kuntze)by using the RACE-PCR technique(designated as CsHMGR1). It comprised 1 979 bp,with a 1 722 bp intact open reading frame encoding a 573-amino-acid protein. The deduced protein showed 80% to 82% similarities to homologs from rubber tree(Hevea brasiliensis),common camptotheca fruit(Camptotheca acuminate),ginseng(Panax ginseng),litchis(Litchi chinensis),American ginseng(Panax quinquefolius),rooted salvia(Salvia miltiorrhiza),Momordica grosvenori(Siraitia grosvenorii),and longan(Dimocarpus longan). The phylogenetic tree,constructed with the catalytic domained of CsHMGR1 and homologs from other species,indicated that CsHMGR1 belonged to the eukaryotic class I HMGR family. CsHMGR1 consisted of two transmembrane domains,implying that it may be localized to endoplasmic reticulum(ER)similarly to other eukaryotic homologs. It also contained two HMG-CoA binding sites,two NADP(H)-binding sites,four conserved catalytic active residues and a phosphorylation site,indicating that phosphorylation/dephosphorylation is likely a crucial mode of regulation of its biochemical activity. Tissue expression analysis indicated that CsHMGR1 was expressed comparatively in the leaf buds of C. sinensis cv. Dayelong and in both leaf buds and floral buds of the mother plants. The regulation of expression and physiological activity of CsHMGR1 are likely to impact greatly on tea quality,and CsHMGR1 may provide a basis of the quality evaluation and breeding of tea given that its function is further resolved.]]> 2015/12/14 17:14:58 1, XU Ling-Ling^1*, LIAO Liang¹, LI Tong-Jian¹, DENG Hui-Sheng¹, FAN Qi-Shui², XU Xiao-Qing³]]> HAN Xing-Jie¹, XU Ling-Ling^1*, LIAO Liang¹, LI Tong-Jian¹, DENG Hui-Sheng¹, FAN Qi-Shui², XU Xiao-Qing³ 17true Torenia biniflora is a member of Scrophulariaceae Torenia,and it is an annual flowering plant that widely grows in Chinese subtropical area,such as Guangdong Province and Guangxi Zhuang Autonomous Region. T. biniflora is a wild flower with high great ornamental value. However,in natural growth condition,the reproduction speed of T. Biniflora is slow,proliferation rate is low,and the kinds of flower colour and type are few,therefore,it is unable to meet the diversified need of market. Plant tissue culture technology provides a new way to improve and breed new varieties for ornamental plants. Up to now,a lot of members of Torenia plants have been successfully established in vitro regeration system through tissue culture,such as T. fournieri Linden,T. baillonii Susie Wong and T. concolor Lindl.,etc. While,tissue culture for T. Biniflora has never been reported. The main objective of this research was to establish in vitro regeneration system of T. Biniflora through tissue culture and study the effects of plant growth substances such as IBA,6-BA and NAA for clonal multiplication. In this study,we created a reliable protocol for regenerating T. biniflora plants in tissue culture by using leaf explants. The effects of different concentrations and combinations of plant growth substances on adventitious bud induction and growth were studied,and the effects of different ionic strengths of media and concentrations of IBA on rooting were analyzed. According to a series of indices,such as average number of buds,frequency of bud induction,frequency of rooting,average number of root and average length of root,the optimal media for inducing adventitious buds and rooting were screened respectively. The results showed that adventitious bud induction was related with the concentrations and combinations of plant growth regulator,and the optimal medium for buds induction was MS+6-BA 0.5 mg·L^-1+NAA 0.2 mg·L^-1. The best medium for rooting was 1/2MS as basal medium with IBA 0.05 mg·L^-1. In this study the in vitro plant reproductive system of T. biniflora with high frequence was completely established,and it would provide an important experiment foundation for the research of rapid propagation and genetic transformation for T. biniflora.]]> 2015/12/14 17:14:59 1, SHI Qiu-Ying¹, CHEN Xiong-Wei¹, CHEN Gang^1*, JIN Hong²]]> WANG Ying-Hua¹, SHI Qiu-Ying¹, CHEN Xiong-Wei¹, CHEN Gang^1*, JIN Hong² 16true LDC gene,the cDNA of LDC was cloned by RT-PCR technique from chilling tolerant ‘Chipper' cucumber treated under low temperature. The sequence of amino acids and conserved sequence of protein were analysed with software DNAMAN6.0 and by BLASTp. A plant expression vector pCAMBIA1304-LDC was constructed by the methods of double restriction enzyme digestion and T4DNA ligase. The selective concentrations of Hyg for adventitous buds induction and shoots growth were confirmed by adding differents concentration of Hyg to the media,and the infection condition of agrobacterium was optimized. The results showed that the coding sequence of the LDC gene had 648 bp in length,encoding 216 amino acids which contained lysine decarboxylase conserved sequence,so the sequence was firmly believed as LDC gene. The accession number was KC202438. The induction rate of adventitious buds reduced to 5.41% when the concentration of Hyg was 10 mg·L^-1,and the survival rate was significantly lower than control group when adding 20 mg·L^-1 Hyg into medium. The optimized transformation condition was no preculture,the suitable concentration OD=0.6,infection for 5 min,and coculture duration 4 d. and higher resistant buds differentiation rate were gained under the optimized condition. 29 T0 plants were identified by PCR technique on DNA level,the transformation rate reached 93.55%. The gene modified plants would be used for the research of LDC gene function in future.]]> 2015/12/14 17:15:00 1, MIAO Yong-Mei^2*, SUI Yi-Hu², BIAN Zhuo-Wu², CHEN Cun-Kuan², HUANG Chun-Jing², LI Zhen-Xing²]]> JIAN Xing¹, MIAO Yong-Mei^2*, SUI Yi-Hu², BIAN Zhuo-Wu², CHEN Cun-Kuan², HUANG Chun-Jing², LI Zhen-Xing² 15true ygl-63 was identified, which derived from Nipponbare(Oryza sativa ssp. japonica)treated by ethyl methanesulfonate(EMS). The mutant ygl-63 exhibited distinct yellowish green leaf trait throughout the growth period. To characterize the yellowish green leaf phenotype of the ygl-63 mutant, we measured its chlorophyll contents at the seedling stage. Compared to that of its wild-type parent Nipponbare, the content of chlorophyll(Chl)a, Chl b and total chlorophyll decreased significantly in the mutant ygl-63, with 31.9%, 42.2% and 34.1% respectively, indicating that the mutant phenotype of ygl-63 was resulted from reduced chlorophyll level. In addition, the ratio of Chl a/b was increased, due likely to the potential of Chl b synthesis in suffering a more severe decline than Chl a in the ygl-63 mutant; and at maturity, the number of productive panicles per plant and seed setting rate reduced by 8.9% and 8.5%, respectively; the 1 000-grain weight increased by 10.4%; but the plant height, panicle length and the number of spikelets per panicle were not affected remarkably with its wild-type parent Nipponbare. Meanwhile, by measuring the contents of micronutrients, we found that the Fe and Zn contents in the seeds of ygl-63 mutant were significantly reduced by 85.7% and 64.8% respectively, compared with its wild-type parent Nipponbare. Genetic analysis of F₁ and F₂ generations of ygl-63 mutant crossed with the normal green variety Minghui 63(Oryza sativa ssp. indica)showed that the mutant trait of ygl-63 was controlled by a single recessive nuclear gene. Genetic mapping of the mutant gene was conducted by using SSR and InDel molecular markers and 166 F₂ plants from the cross of ygl-63 with the normal green variety Minghui 63, and the mutant gene of ygl-63 was finally mapped on the long arm of rice chromosome 11. The genetic distances from the target gene to the markers InDel-3 and InDel-5 were 0.9 and 1.5 cM, respectively. The gene ygl-63 was considered to be a new rice yellowish green leaf mutant and its mutant gene was tentatively named as ygl-63(g). These results will provide the information for the cloning and functional analysis of ygl-63(g)gene in the future.]]> 2016/9/3 0:58:27 1, ZHANG Fan-Tao^1,2*, NIE Li¹, WAN Ling-Fei¹, LIANG Jian-Qiu¹, ZHANG Yu-Jia¹, MA Yang-Shuai¹, XIE Jian-Kun¹]]> ZHANG Liang-Xing¹, ZHANG Fan-Tao^1,2*, NIE Li¹, WAN Ling-Fei¹, LIANG Jian-Qiu¹, ZHANG Yu-Jia¹, MA Yang-Shuai¹, XIE Jian-Kun¹ 14true Nanophyton erinaceum has a long living history at Aletai Copper Mine in Xinjiang. In order to analyze the relation between metallothionein gene and heavy metal response stress, a new heavy metal-responsive gene cDNA sequence was successfully cloned by RACE(Rapid amplification of cDNA ends)from N. erinaceum. The gene was named as NeMT2, and its accession number in GenBank was KT835290. The metallothionein gene NeMT2 was 590 bp in full length, and it had 237 bp ORF(open reading frame)which encoded 78 amino acid residues. There were 14 Cys residues, arranged in the form of C-C, C-X-C and C-X-X-C, in the 78 amino acid residues, and these Cys residues distributed in N terminal and C terminal of peptides. The protein encoding by gene NeMT2 molecular weight was 7.603 6 kD and its isoelectric point was 4.71. Phylogenetic analysis results demonstrated that the deduced amino acid sequence of gene NeMT2 was the highest homology as the Salicornia brachiata(AEF01492)and the Halostachys caspica (AHI62953), secondly the Beta vulgaris (XP_010667708.1), but the lowest similarity with the Nelumbo nucifera (XP_010253171). Bioinformation analysis showed that metallothionein NeMT2 had no signal peptide and belonged to the hydrophilic non-transmembrane protein. Hydrophobicity analysis was carried out and the results showed that there was a strong hydrophobicity region between 35 to 45 amino acids, and the 41 Asp had the strongest hydrophobic property(1.444). Predication of structure indicated that random coil was the major components of its secondary structure. Expression analysis of NeMT2 gene was carried out by RT-PCR. The results showed that expression of NeMT2 gene was detected both in Copper Mine and nor Copper Mine in Nanophyton erinaceum leaves, but the former was more stronger than the latter, which indicated that NeMT2 gene was responsive to the heavy metal stress. Then NeMT2 gene in Nanophyton erinaceum was cloned into 35S promoter downstream of plant over-expression vector pCAMBIA1300 in orientation. The plant overexpression vector pCAMBIA1300+NeMT2 was successfully constructed. These findings will provide information for the functional study of NeMT2 and its molecular mechanism in repnse to the heavy metal stress.]]> 2016/9/3 0:58:27 *]]> GE Feng-Wei, ZENG Wei-Jun, LI Yan-Hong, ZHAO Hui-Xin^* 13true -1, for 21-ES was 1.67 mL·L^-1 respectively. The analysis of biological effects in M_l generation showed that the variation rangs and variation coefficients of the different traits were improved, and the biological effects of different traits had different performances in the three inbred lines, which indicated that the different traits of the different materials had variable sensitivities in the different concentrations of EMS, sensitivity of the growth period was 21-ES>K305>R08, sensitivity of the plant type traits was R08>21-ES>K305; sensitivity of tassel traits was K305>21-ES>R08, sensitivity of the main ear traits was 21-ES>K305>R08. The analysis of mutagenic effects in M₂ generation showed that general mutation spectrum was expanded, variation of plant height, ear height, leaf area and the main ear characters performed widely. The main tassel traits were two-way variation except for branch number in K305 M₂ strains. This study provides the foundation for the further research and application.]]> 2016/9/3 0:58:27 1, CHEN Chao-Jie^1,3, XIA Wei², YU Xue-Jie¹, KE Yong-Pei^1,2*]]> SHI Hai-Chun¹, CHEN Chao-Jie^1,3, XIA Wei², YU Xue-Jie¹, KE Yong-Pei^1,2* 12true Kobresia tibetica, belonging to the genus of Kobresia and the family of Cyperaceae, is a perennial herb. The rhizomes of K. tibetica are short and culms are densely tufted. It is mainly distributed in Qinghai, Gansu, west of Sichuan provinces and east of Tibet Autonomous Region at altitudes ranging from 2 550 m to 4 950 m and can be found in alpine swampy meadows, weedy plains, marshes and riversides. The root system of K. tibetica is developed. Besides, it is hygrophilous, resistant to cold and strong in vitality. For the reproduction of K. tibetica, vegetative propagation is prior to sexual propagation. K. tibetica plants are of highly ecological and economic importance. The stem and leaf of K. tibetica are luxuriant. K. tibetica contains high levels of crude proteins and fats and is preferred by livestock over other plants found on the plateau. The production of K. tibetica grass is of high yield. As a consequence, it is the major grazing forage grass of summer and autumn in Qinghai-Tibet Plateau. Furthermore, K. tibetica is the constructive and dominant species of alpine swamp meadows in Qinghai-Tibet Plateau. K. meadows are also highly involved in carbon storage and serve as important carbon sinks in China. Therefore, K. tibetica was chosen as the research object in this study. In order to establish the suitable Inter Simple Sequence Repeats Polymerase Chain Reaction(ISSR-PCR)system in K. tibetica, factors which affect the ISSR-PCR amplification, such as magnesium ion(Mg²⁺), Taq DNA polymerase, dNTP, primer and DNA template, were optimized and selected by orthogonal designed experiment of five factors in four levels. In addition, suitable ISSR primers and the annealing temperature of selected primers were screened from 100 ISSR primers by gradient PCR. As a result, the optimal reactions of ISSR-PCR in K. tibetica were performed in a 20μL volume containing 2μL 10× PCR buffer, 1.5 mmol·L^-1 Mg²⁺, 1.0 U Taq DNA polymerase, 0.100 mmol·L^-1 dNTP, 0.3μmol·L^-1 primer and 30-40 ng template DNA. Out of 100 ISSR primers screened, twelve primers were selected with clear and steady bands and suitable annealing temperature of each primer was 48.0-53.2 ℃, depending on primers used. The appropriate program of ISSR-PCR for K. tibetica was as follows: an initial step of 5 min at 94 ℃ followed by 38 cycles of denaturing at 94 ℃ for 20 s, annealing at appropriate annealing temperature 48.0-53.2 ℃(different primers with different annealing temperatures)for 1 min and extending at 72 ℃ for 80 s, ending with a final extension of 6 min at 72 ℃. The stability of the ISSR-PCR reaction system indicated that clear and rich polymorphism bands were obtained in different individuals of K. tibetica with the optimal system. The establishment of ISSR-PCR system was beneficial to the analysis of genetic diversity and genetic structure of K. tibetica germplasms. This study will provide the information for further study of screening high quality forage. Furthermore, it is of great significance for the study of Kobresia-swamp meadows in Qinghai-Tibet Plateau as well as the restoration and conservation of the wetland ecosystem.]]> 2016/9/3 0:58:27 1, BAO Rui¹, WANG Li¹, SHI Lin^{1, 2}, LI Yi^1*]]> HU Yan-Ping¹, BAO Rui¹, WANG Li¹, SHI Lin^{1, 2}, LI Yi^1* 11true Cinnamomum longepaniculatum. The results showed that four kinds of fungal endophytes had obvious inhibitory effects on C. longepaniculatum cell growth, and the denser the fermentation fluid was, the stronger inhibitory effects they had. The trend of the effects that the four kinds of endophytic fungi had on the total volatile of secondary metabolites accumulation and C. longepaniculatum oil component 1,8-cineole γ-terpinene and α -terpineol accumulation in suspension cultures of C. longepaniculatum was promoted at a low concentration and inhibition at a high concentration. The promotion effects of 1% YG42, YY26 and 0.25% YY11 on the total volatile of secondary metabolites accumulation were equivalent, and they had the strongest promoting effects on the total volatile of secondary metabolites accumulation, which were 2.00, 1.95 and 2.01 times of the control respectively. The 0.25% YG71 had the strongest promoting effects on the 1,8-cineole accumulation, which was 11.03 times of the control. The 0.25% YG71 and YY26 had the strongest promoting effects on the a-terpineol accumulation, which were 1.72 and 1.81 times of the control respectively. γ-terpinene was not detected in the control. Four kinds of endophytic fungi could induce γ-terpinene production, the biggest peak areas were 0.19, and the endophytic fungi was 0.25% YG71. This research will enrich the theory that fungal endophytes affect the volatile of secondary metabolites in aromatic plant, and provide experimental evidence for the use of endophytic fungi to increase the amount of useful component in C. longepaniculatum oil accumulation in production.]]> 2016/9/3 0:58:27 1, TAN Yun-Ya², LI Qun^2*, YOU Ling¹, WANG Chao², WANG Yu¹, LIAO Lin¹]]> WEI Qin¹, TAN Yun-Ya², LI Qun^2*, YOU Ling¹, WANG Chao², WANG Yu¹, LIAO Lin¹ 10true -1 in MS media. The percentage of callus induction did not changed when NAA was added in basic media in callus induction process. The results showed that it was not helpful for callus induction when NAA was added in MS basic media. Four treatment combinations including two different concentrations of 2,4-D and 6-BA were used to subculture for induced callus. The best combination was treatment No. 1, which included 1 mg·L^-1 2, 4-D and 0 mg·L^-1 6-BA. When 6-BA was added in MS basic media, callus became brown and the growth of callus was also inhibited. These results indicated that 6-BA inhibited the growth of callus in subculture process when 6-BA was added in MS basic media. To gain a better understanding about the effects of different combinations of plant growth substances in callus differentiation, five treatment combinations including two different 6-BA and NAA concentrations and three different concentrations of KT were used to differentiate for callus when the callus had been subcultured. Treatment No.1 was the best combination in the five treatment combinations. And the differentiation rate of callus from mature seeds of S722 was 33.3% when 2 mg·L^-1 6-BA was added in MS basic media. It was the highest differentiation rate when the concentration of 6-BA was added in media. Callus from S722 mature seeds were more easily differentiated than the other sudangrass strain Sa. Summarily, the best concentration of 2,4-D for callus induction and subculture was 1 mg·L^-1 in basic media when mature sudangrass seeds were used for explants. 6-BA had inhibition effects for callus in subculture process. And the best concentration of 6-BA for callus differentiation was 2 mg·L^-1 in basic media. The different sudangrass strains should use different combinations of plant growth substances in tissue culture process. Therefore, plant regeneration rate would be improved if the concentrations and combinations of plant growth substances were adjusted in media for tissue culture and plant regeneration of sudangrass when different sudangrass strain's mature seeds were used as explants.]]> 2016/9/3 0:58:27 LI Jie-Qin, WANG Li-Hua, ZHAN Qiu-Wen, HU Neng-Bing, WANG Shi-Jian 9true Salvia prionti Hance is a famous genuine medicinal materials in Guangxi and commonly ustilized in traditional Chinese medicine. It has strong antibacterial activity and anticancer effects and mainly used to cure bacillary dysentery, diarrhea, enteritis, pneumonia, acute pharyngitis, tonsillitis, colds, etc. With the discovery of new effective components and expansion of medicinal range, S. prionitis has become a promising variety in the field of traditional Chinese medicine. In order to rapidly identify and evaluate the differences of chemical composition in S. priontis which from different original locations, we combined with the principal component analysis(PCA)and cluster analysis as well as loading factor analysis, and used the fourier transform infrared spectroscopy(FTIR)to determinate the samples of S. priontis from different original locations, so that it could be effective to find out the main chemical composition information of PCA cluster variations and rapidly authenticate the quality of different samples. The results indicated as follows:(1)The fingerprints of bands from 1 800 to 600 cm^-1 showed that the all samples of S. priontis had similar absorbance bands such as 1 727,1 635,1 551, 1 513, 1 442, 1 373, 1 255, 1 154, 1 036, 795, 776, 690 cm^-1, which indicated that the chemical compositions of S. priontis in different original locations were still relatively stable.(2)Based on the vibrational characteristics of FTIR fingerprints in differernt samples, the classification of principal composition and cluster analysis results showed that the relationship of chemical component of each S. priontis had significant correspondence with their geographical location and environment climatic conditions. In the near class, the chemical components were similar to each other, on contrary, the chemical component of S. priontis among in far class had obvious differences. So these two methods were both able to quckly identify S. priontis in different original locations, while, the two methods had individual characteristics.(3)According to the PCA loading factor analysis, more chemical components could be out found compared to the orgianl FTIR fingerprints among the different detected samples, and the absorption bands could also be quickly found out, which were significant contributed to the classification of principal components and cluster analysis. Among all the absorbtion bands, the bands arounds 1 670, 1 630, 1 616, 1 579, 1 473, 1 411, 1 159,1 129, 1 082, 1 042, 1 000, 972, 946, 913, 891, 806 cm^-1 were obviously correlated to the classfication of PCA, among which, five bands were from saprorthoquinone and tanshinone, six bands were from prioket-olactone, sterols components. Therefore, the differences of FTIR fingerprints in S. priontis from different original locations were mainly due to the differences of some chemical composition and concentration including prioketolactone, sterols components, saprorthoquinone and tanshinone. The method of this resarch is simple, quick and undamaged, and can be used to quickly identify and evaluate the quality of S. priontis from different original locations. At the same time, the present study will provide reference for cultivation and well-bred breeding work of S. Priontis.]]> 2016/9/3 0:58:27 *, WANG Man-Lian, ZOU Rong, SHI Yan-Cai]]> KONG De-Xin, TANG Hui^*, WANG Man-Lian, ZOU Rong, SHI Yan-Cai 8true Oxytenanthera abyssinica is the most important indigenous bamboo in Africa. It plays an important role in local life, production and environment protection. However, the study of its genetic variation is little. We studied seed characters and its variation, as well as the genetic variation of its seedlings abide by GB2772-1999. The length of seeds was 18.2 mm, width was 3.3 mm, 1 000 seed mass was 117.5 g and germination percentage was 66.5%. The results showed that little variation occurred in its seed characters, and variation degree was similar to that of other bamboos. More than 15 000 seedlings was cultured, leaves of 64 individuals were chosen as test material typically, 39 of them were chosen randomly and 25 individuals with big leaves. AFLP was applied to test its genetic variation, eight pair of primers are used. After electrophoretic separation, 1 728 lines was calculated by GeneScan 3.1. It occurred that its PPB was 90.28%, Nei's gene diversity index was 0.243 6, Shannon index was 0.362 3. The result showed that the genetic variations was abundant in O. abyssinica. Based on simple matching coefficient(SM), the individuals were clustered by UPGMA cluster analysis. It indicated that the SM index ranged from 0.73 to 0.85. The samples can be clustered into three classes when SM similarity coefficient is 0.76. Combine phenotypic and physiological indicators systematic germplasm exaviation is further needed. This study will provide scientific information for biodiversity breeding and improved breeds.]]> 2016/9/3 0:58:27 1, ZHANG Guo-Wu², FENG Yun¹, RAN Hong¹, ZHANG Ying¹, GUO Qi-Rong^1*]]> LIAN Chao¹, ZHANG Guo-Wu², FENG Yun¹, RAN Hong¹, ZHANG Ying¹, GUO Qi-Rong^1* 7true Cymbidium cyperifolium var. szechuanicum and C. floribundum were studied by using ISSR marker. Fourteen ISSR primers were selected from 80 ISSR primers, which were used to amplify 61 individuals including paternal, maternal and 59 F1 progeny, 107 DNA fragments were produced by using these ISSR primers to amplify, every primer had 7.64 bands in average. The length of amplified DNA fragments ranged from 90 bp to 2 100 bp, and there were 11 kinds of bands. The results showed that the polymorphism of ISSR marker in the F1 progenies was high, and the segregation locus was 44.86%. And 83.33% segregation loci accorded with the segregation patterns of Mendelian segregation model with the segregation ratio nearly 1:1 or 3:1. 12.50% segregation loci deviated from Mendelian segregation model, 4.17% segregation loci belonged to special bands. Six deviated from Mendelian segregation model bands, eight deficiency bands and two new genetic character bands would cause some new variation. Cluster analysis showed that the genetic distances of both parental and 59 F1 progeny is further, fifty-nine F1 progeny in one group, then fifty-nine F1 progeny and female plant were in one group, at last 59 F1 progeny and female plant and male plant were in one group, this result indicated that 59 hybrids were partial female type. The hybrids of Cymbidium cyperifolium var. szechuanicum and C. floribundum had some common characteristics of their parents, such as leaves, chromosomes and hereditary material. Sixty-one materials were identified by ISSR markers and morphological characters that 59 F1 progeny obtained the genetic characteristics of Cymbidium cyperifolim var. szechuanicum and C. floribundum, so they were true hybrids. Fifty-nine F1 hybrids had mutually complementary bands of both parents, the F1 hybrids also produced some new specific bands which were recombinant fragment of parents. This experiment showed that 59 F1 progeny had both genetic and variation. Therefore, ISSR markers can be used as an effective molecular technique for directional selection of the interspecific hybrid and the study of orchid cross-breeding.]]> 2016/9/3 0:58:27 1,2, HU Chun-Gen^1*]]> ZHOU Li^1,2, HU Chun-Gen^1* 6true 3 is widely used in plant tissue culture. In this study, more than four years old anther callus was used as a material for the research of the appearance and cell morphology changes by pre-culturing on subculture medium supplemented with 2.5 mg·L^-1 AgNO₃. In order to promote the frequency of somatic embryogenesis, different concentrations of AgNO₃ were added to somatic embryo induction medium, and different treatment time with AgNO₃ was tested. Total number of embryoid and number of normal somatic embryos were respectively counted after 90 d. The results showed that the pale yellow soft callus could turn into bright yellow fragile callus after pre-culture on medium containing 2.5 mg·L^-1 AgNO₃. Under an inverted microscope, the former was mostly irregular polygon with relatively thin cytoplasm, and the latter was seen to be sphere or ellipsoid, rich in cytoplasm, which was considered to be typical embryogenic cell. Adding 5 mg·L^-1 AgNO₃ to somatic embryo induction medium during the first month, significantly enhanced somatic embryo production, while 10 mg·L^-1 AgNO₃ inhibited somatic embryogenesis and more abnormal embryos emerged. Development of somatic embryo was obviously hindered when callus was cultured on the induction medium containing 5 mg·L^-1 AgNO₃ for more than 2 m, and most of them were deformity. In conclusion, this study recovered the embryogenic characteristics of long-term subculture anther callus to some extent, and increased the frequency of somatic embryogenesis, which provides references for maintaining embryogenic capability during long term subculture of anther callus in Hevea brisiliensis.]]> 2016/12/26 11:22:52 *]]> DAI Xue-Mei, HUANG Tian-Dai, LI Ji, YANG Xian-Feng, XIN Shi-Chao, HUANG Hua-Sun^* 5true Melasma arvense is known as one of the precious herbs in Yunnan Province, and has the effects on leukemia, cancer, dispersing blood stasis and hemolysis. It has become a hot spot for the development of natural drugs in Yunnan. At present, because of its peculiar habitat, seed dispersal difficulty, reproductive barriers, seed germination and establishment difficulty and over-harvesting, wild plants are becoming extinctive recent years in Yunnan Province. Therefore, in order to realize resource protection and restoration, it is necessary to carry out resource conservation and artificial breeding. In order to primary establish the adventitious bud induction and multiplication system that present excellent properties of growth and multiplication, used stem shoots as tissue culture inoculation materials studied the effects of different medium type, different concentrations of cytokinin and different natural organic additives on bud multiplication and growth based on the prophase research of provenance collection, artificial propagation and screening of different organ culture of M. arvense. The results indicated that about 2-3 cm long stem with buds cultured on cultural conditions(1/2 MS with 6-BA 0.5 mg·L^-1 + Pt 50 g·L^-1 + Bn 80 g·L^-1 + 25 g·L^-1 sugar + 7 g·L^-1 agar with pH=5.8, a temperature of 22 ℃, light 10 h·d^-1)for 8-10 d, they produced numerous adventitious buds. Each explant produced 8.1 shoots and the grow length of main stem was 74.2 mm in 40 d. The experiment primarily established the adventitious bud induction and multiplication system that present excellent properties of growth and multiplication. This study provides a scientific basis for the artificial cultivation and sustainable use.]]> 2016/12/26 11:22:52 1, FANG Hai-Ling², WANG Ding-Kang¹, XIANG Xiao-Dong³, JIN Song¹, GENG Kai-You^1*]]> LI Yu-Chuan¹, FANG Hai-Ling², WANG Ding-Kang¹, XIANG Xiao-Dong³, JIN Song¹, GENG Kai-You^1* 4true Mirabilis jalapa is a perennial herb of Nyctaginaceae, native to South America. It grows faster and can reproduce itself. Not only is it a high-value ornamental flowering and greening plant, but also can be used as medicine. In addition, the recent studies show that Mirabilis jalapa has good bioremediation function. Plant tissue culture technology provides an important way to improve and breed new varieties for horticultural plants. However, the research related to M. jalapa is still relatively little, and there are no reports on plant regeneration of M. jalapa at present. In this study, the achenes of M. jalapa were sterilized and inoculated into basal MS medium containing 0.1 mg·L^-1 NAA for the sterile seedlings obtained. Then the leaves and stems of the sterile seedlings were as explants to study the effects of different plant regulators on the buds induction and plant regeneration. According to frequency of callus induction, average number of buds and roots, the optimal media for inducing adventitious buds and rooting were screened respectively. The results showed that most of leaf explants did not produce calli, none of which formed adventitious buds. Moreover, with the prolongation of culture time, leaves began to produce adventitious roots, and gradually became more and more. The nodal stem segments of vegetative branches of sterile seedlings were more suitable for the induction of buds. Micropropogated shoots were quickly induced from axillary buds of nodes on an induction medium consisting of basal MS medium supplemented with different concentrations of 2,4-D, NAA, KT, 6-BA, and TDZ. Among them, the optimal medium for the explant culture in vitro of M. jalapa was MS + 1.0 mg·L^-1 6-BA + 1.5 mg·L^-1 KT + 1.0 mg·L^-1 NAA + 0.05 mg·L^-1 TDZ, the number of adventitious shoots were more and growth robust. For rooting induction, either MS or 1/2MS medium could induce the adventitious buds to produce roots, and the optimal medium was 1/2MS + 0.5 mg·L^-1 NAA. This study was designed to explore the optimal conditions for tissue culture of M. jalapa, which provides the information for the future establishment of an efficient regeneration system of M. jalapa tissue culture and genetic transformation system.]]> 2016/12/26 11:22:52 1,2,3, ZHANG Han-Jie¹, HAN Wen-Yu³, SHEN Zhi-Qiang^2*]]> LU Yu-Jian^1,2,3, ZHANG Han-Jie¹, HAN Wen-Yu³, SHEN Zhi-Qiang^2* 3true Brassica juncea(AABB, 2n=36)is one of the 3rd Brassica oil rape(the other two oil rapes are: B. rapa and B. napus)in China, the species has many desirable traits, such as strong drought resistance and strong resistance to disease and pest. As the important origin of B. juncea worldwide, China has rich valuable germplasm resources. In this study, we collected 34 B. juncea accessions from all over China, and planted all these accessions with three randomalized replications in Guiyang environment. The fatty acids, including erucic acid, oleic acid, linolenic acid, linoleic acid and stearic acid, was detected by NIR method in this study. All the detected fatty acids showed quantitative and normal distribution in Guiyang environment. Furthermore, we analyzed the correlations among the five above detected fatty acids, the erucic acid and oleic acid are significantly negatively correlated, the linolenic acid and stearic acid are significantly negative correlated, the linolenic acid and linoleic acid are negative correlated. To classify the 34 accessions for better utilization in B. juncea breeding, the principal component analysis(PCA)was used and the results indicated that most of the 34 accessions(30 accessions, about 88.2%)located on one same area, only several other accessions, that was SL63, Lengjiao YC, T6342 and Changyanghuangjie, located on the separate areas. These scattered accessions had special values in B. juncea breeding for their very different traits when comparied with the other accessions. Besides, we BLAST two FAE1 gene sequences in NCBI website from B. napus (AACC, 2n=38)and B. oleracea (CC, 2n=18), respectively, which were further used for designing specific primer for FAE1 gene, responsible for the regulation of erucic acid in Brassica species. The primer showed a good amplyfication in the all the 34 B. juncea accessions. So, this study proves that the B. juncea genome contains at least one FAE1 copy. In conclusion, the study detected the fatty acid content of 34 B. juncea accessions in Guiyang environment and also provides the useful FAE1 primer for the future gene clone in B. juncea, all of which are valuble in the future B. juncea molecular breeding of China.]]> 2016/12/26 11:22:52 1*, LI Lu-Feng², JIA Shi-Yan³, LIN Shu-Chun¹]]> TIAN En-Tang^1*, LI Lu-Feng², JIA Shi-Yan³, LIN Shu-Chun¹ 2true Prunus domestica cv. France, using P. domestica ‘France' as female parent, ‘Stanley' and ‘Empress' as maleparents, and made an overall evaluation according to principle components. Among them, ‘France' has a brittle fruit, sweet flavor characteristics welcomed by consumers. But because the ‘France' varieties of fruit smaller, would produce a certain gap between the yield and fruit quality aspects of the current market demand, restricting the rapid development of industry prunes. Therefore, a clear prunes pollination characteristics, production rational selection and configuration prunes pollinated varieties is an important measure to improve the yield and quality. The results showed that different varieties pollinating had a certain role in improving fruit setting rate of P. domestica ‘France', in which, fruit setting rate of Empress was higher than that of others. There were significant differences on main economic characters of fruit such as vertical wrap of nuts, vertical wrap of stone, stone weight, vertical wrap of fruit, diameter of fruit, water content, soluble sugar, soluble protein, Vitamin C. However, there was no obvious difference on the diameter of nuts, nuts weight, stone weight, stalk of fruit, separation stone, skin color, flavor, soluble solids, hardness, total acid. Five principal components whose total cumulative contribution reached 90.694% were extracted by principal component analysis, and the formula of quality evaluation of pollination varieties was as follow: F=6.088F1 + 3.964F2 + 2.406F3 + 1.666F4 + 1.294F5. Comprehensive scores of different pollinated varieties could be caculated and sorted with this model. They were between -1.263 to 1.260, from high to low, the arranging order of the comprehensive scores of the different pollination varieties were assessed by the comprehensive evaluation function: ‘Empress' > ‘Stanley' > ‘Natural' pollination. Empress pollination is better through comprehensive comparison. This experiment provides a reference for quality improvement optimize the allocation of pollination tree of P. domestica ‘France'.]]> 2016/12/26 11:22:52 1, TIEMUER Ayiguli¹, TUOHETI Bilikezi², REYIMU Miriban², KADIER Rexiati²]]> ABULA Nuerman¹, TIEMUER Ayiguli¹, TUOHETI Bilikezi², REYIMU Miriban², KADIER Rexiati² 1true