-->Genetics and Breeding

Comparative analysis on codon usage bias in mitogenome of two species in genus Glycine

2020/8/17 12:39:59

The characteristic of codon usage in mitogenome of wild and cultivated soybean was studied and compared to explore the main factors affecting codon usage bias and codon evolution. The results were as follows:(1)The GC content in the coding region of mitochondrial genome of cultivated soybean and wild soybean was 44.56% and 44.58%, respectively, which indicated that the coding sequences of soybean mitogenome was rich in A/T base.(2)PR2-plot analysis showed that the third base of codon usage was biased, where pyrimidine was used more frequently than purine; Nc-plot analysis revealed that the number of genes with Nc ratio in the range of -0.1-0.2 accounts for more than 95% of the total number of genes. The above-mentioned results showed that the multiple factors such as mutation and selection affected the formation of codon usage bias in soybean mitogenome. GC1 and GC2 value was significantly correlated with GC3, suggesting that codon bias in the mitogenome of cultivated and wild soybeans was mainly affected by mutation.(3)In addition, 20 and 21 codons were identified to be the optimal codons for the mitogenome of cultivated and wild soybeans, with most of the preferred synonymous codons ending with A or T base. The formation of mitochondrial codon bias of cultivated soybean was more affected by selection than that of wild soybean, which may be the result of long-term artificial cultivation and domestication of cultivated soybean.

Cryopreservation of sterile Pinus massoniana seeds

2020/8/17 12:39:59

To explore long term preservation of Pinus massoniana seeds through cryopreservation technology, aseptic seeds of P. massoniana with different water contents(WC)(27.4%, 24.6%, 22.7%, 16.8%, 15.8%, 10.7%, 7.5%, 6.1%, 4.8%, 3.2%)were cryopreserved in liquid nitrogen. Our results showed that:(1)The germination of cryopreserved seeds increased when the seed WC elevated from 3.2% to 6.1%, reaching the highest rate of 91.33% at the optimum WC of 6.1%; However, seed germination decreased as WC further increased from 6.1% to 27.4%, in particular, with a rapid decline in germination of seeds with 15.8% or higher WC; There is a wide range of safe WC, within 3.2%-7.5%, the average germination rate reached above 80%.(2)In addition to WC, freezing and thawing methods also affected cryopreservation. Rapid freezing by directly immersing seeds into liquid nitrogen is better than pre-freezing at -20 ℃ prior to liquid nitrogen freezing; a slow air-thawing at room temperature gave a better germination result compared with a rapid thawing in a 42 ℃ water bath that caused seed episperm cracking.(3)The episperm also played an important role, which protected the seeds from the mechanical damage. The germination rates without episperm were significantly lower than those with episperm, and the abnormal seedlings were more observed when the episperms were removing.(4)At the optimum WC of 6.1%, cryopreserved seeds germinated better than those non-cryopreserved(control), indicating that cryopreservation may stimulate seed germination of P. massoniana. Collectively, our results showed that water content, freezing and thawing methods as well as the episperm structure can significantly affect cryopreservation of P. massoniana seeds; and the best method for cryopreservation of P. massoniana seeds is to directly freeze seeds with 6.1% WC using liquid nitrogen followed by a slow air-thawing at room temperature, by which the germination can reach above 90%. Results from this study may provide a rational for a long-term storage of P. massoniana seeds through cryopreservation.

Effect of simulated drought stress on seed germination and geographical variation of six provenances of Catalpa ovata

2020/8/17 12:39:59

In this study, polyethylene glycol(PEG-6000)was used to simulate drought stress, the germination percentage, germination potential and radicle length were measured. Correlation analysis, cluster analysis and membership function method were used to comprehensively evaluate six provenances of Catalpa ovata. The most resistant sources of drought resistance were screened good rootstocks out for the grafting of Chinese C. bungei, to promote the breeding, and to lay a material foundation for its large-scale application. The results were as follows: The germination rate, germination potential, relative germination rate, germination index and vigor index of six provenances of C. ovata seeds decreased with a trend continuous decline. Except for the radical length, hypocotyl length of the Luoyang in Henan provenance, and the radical length of the Zhengning in Gansu provenance. The length of radicle and hypocotyl of other provenances showed a trend of gradual decrease. When the osmotic stress was -1.0 MPa, the Luoyang in Henan provenance was deactivated, and the indexes of another provenances' tendency to be consistent. The correlation between the germination characteristics of each group and the geographical environment factors of origin indicated that there was a significant positive correlation between vigor index, longitude and latitude(0.903 and 0.871). The higher average annual temperature, the lower seed germination rate and vigor. Annual precipitation was negatively correlated with seed germination rate and vigor index, but moderately positive correlated with the growth of radicle and hypocotyl. Temperature and precipitation affected the drought resistance of seeds of C. ovata to some extent, mainly reflected in the lower annual temperature and annual precipitation, the lower germination rate of seeds. It was preliminarily found that provenances of C. ovata in the relatively harsh environment of drought and cold showed the regulation mechanism to improve seed germination rate and reduce the growth activity of seed embryo to adapt to the unfavorable environment. The results of cluster analysis and membership function method were used to comprehensively evaluate the drought resistance of C. ovata seeds in six groups. The seeds of Liaoning in Hengren provenance had the strongest drought resistance, while the Luoyang in Henan provenance had the weakest drought resistance. In summary, the drought resistance of six provenances of C. ovata is obvious, temperature and rainfall affect the drought resistance of seed germination to some extent. The provenance with strong drought resistance are selected to be the rootstocks of C. bungei breeding and lays a material foundation for the large-scale application of C. bungei.

Cracking types of common buckwheat pericarp and its effect on early germination traits

2020/8/17 12:39:59

Common buckwheat, a traditional edible and medical plant, which grain is an important storage organ. However, types of husk-cracking of commom buckwheat and it's germination effects is a scientific and technological problem to be solved. In the present study, five samples of common buckwheat were used as materials. The grain model of common buckwheat was established and the types of pericarp cracking were classified; at the same time, through the paper bed germination method, testing the seed germination rate, the radicle and hypocotyl length. The results were as follows:(1)There are five kinds of cracking types in the grain of common buckwheat, which are longitudinal complete dehiscence type(LCD), semi-dehiscent micropyle(SDM), semi-dehiscent mid-ridge(SDMR), semi-dehiscent hilum(SDH)and transverse cracking type(TC);(2)Five test common buckwheat pericarp cracked type germinates about 12 hours earlier than the pericarp intact type;(3)The early germination rate of common buckwheat varieties with pericarp cracking was higher than that of the corresponding pericarp intact type, however, 48 h later, the germination rate of seeds with intact pericarp continued to increase, and exceeded that of all pericarp cracked types;(4)On the extension characteristics of radicle and hypocotyl, although the growth rate of radicle and hypocotyl of pericarp cracking type were faster than that of intact type of pericarp, the ratio of radicle to hypocotyl of intact pericarp type was higher than that of pericarp cracking type;(5)Different type of pericarp cracking had different effects on seed germination, from big to small, which is SDM, SDH, LCD, SDMR and TC, respectively. Different type of pericarp cracking had different mildew rate, from big to small, which is SDH, LCD, SDMR, SDM and TC. There are five common cracking types in common buckwheat grain pericarp; although pericarp cracking can improve the early germination rate of common buckwheat seeds and promote the early extension rate of radicle and hypocotyl, it can reduce the germination rate and the ratio of radicle to hypocotyl growth of common buckwheat as a whole, it may not be conducive to the formation of strong seedlings and whole seedlings in the field.

Cloning and expression analysis of FmPHV gene in callus of Fraxinus mandshurica

2020/8/17 12:39:59

PHAVOLUTA(PHV)transcription factor is one of the most important members of homeodomain-leucine zipper Ⅲ(HD-ZIP)family, and plays an important role in the regulation of formation of plant stems apical and germ meristem. In this research, the PHV gene of Fraxinus mandshurica was acquired by gene cloning and was named by FmPHV. Detailed bioinformatics analysis showed that the length of FmPHV gene was 2 112 bp, and contained a complete Open Reading Frame(ORF)which encoded a protein with 703 amino acids. The FmPHV gene encoded a stable hydrophilic protein, the subcellular localization prediction of FmPHV protein showed the FmPHV was mainly present in the chloroplast. Conserved domain and homology analysis of FmPHV protein indicated that, the homology of FmPHV between the homology PHV proteins in other species, such as Olea europaean, Sesamum indicum and Nicotiana tabacum, was 99%. Under 4 ℃ condition, with the treatment of 50 mg·L^-1 3-indolebutyric acid(IBA)solution, we obtained the cambium callus from cambium stem cells of Fraxinus mandshurica. Analysis of the gene expression pattern of FmPHV showed that the FmPHV was the highest expression in June. Furthermore, the expression of FmPHV in buds was stronger than which in any other tissues. Compared with the callus from different sources, which obtained from the bark of F. mandshurica, the expression level of FmPHV gene in callus, derived from meristem formation, was significantly higher than which from other sources. Moreover, overexpression the FmPHV gene in F. mandshurica seedlings will reduce the expression of auxin-related genes and increase the expression of cytokinin-related genes. This phenomenon indicated that the function of FmPHV gene was more beneficial to the process of plant shoot differentiation and regeneration. In summary, this research revealed the expression pattern of FmPHV gene during the growth and development in F. mandshurica. And revealed the expression pattern of key genes in plant shoot regeneration pathway, which were regulated by FmPHV gene. Our research reveals the potential value of the molecular characteristic of FmPHV gene in the response of auxin and cytokinin pathways during the process of the formation of plant callus generation.

Genome-wide identification and analysis for oil palm WRKY transcription factors

2020/8/17 12:39:59

Genome-wide sequence information of oil palm from NCBI website, and the WRKY transcription factor sequence from The Arabidopsis Information Resource(TAIR)Database were downloaded, BLAST homology sequence alignment analysis was carried out in oil palm genome database. Protein structure and analysis were carried out through CDD and PFAM database of NCBI online tool, and the series without WRKY domain were eliminated. Bioinformatics analysis and functional prediction of WRKY transcription factor in oil palm. The results were as follows:(1)A total of 95 EgWRKY transcription factors were excavated, and they encoded 116-1 303 bp amino acids, predicting hydrophilic and unstable(except for EgWRKY25 and EgWRKY56). The main structure of 60 EgWRKY proteins was α-helix, and the remaining 35 proteins were irregular curl.(2)The phylogenetic analysis on conserved domain showed that EgWRKY transcription factor family proteins were divided into three categories(I, Ⅱ and Ⅲ). Category I could be separated into I C and I N, and Category Ⅱ was classified into Ⅱ a, Ⅱ b, Ⅱ c and Ⅱ d.(3)The intron-exon structure analysis revealed that structures of EgWRKY gene were highly conserved. This research will lay a foundation for the study of WRKY transcription factor exacavation, function analysis, and molecular biology of oil palm and provide references for its genetic modification and molecular breeding.

Molecular isolation and tissue expression analysis of alternative oxidase family genes in Liriodendron chinensis

2020/8/17 12:39:59

Alternative oxidase(AOX), a terminal oxidase located in respiration electron-transport pathway, which is widely existed in higher plants and closely related to plant respiration. It has shown that AOX played a substantial role in plant growth, seedling morphogenesis and environmental adaptability in recent researches. Liriodendron chinensis is an excellent tree species for garden ornaments and timber use, while the adaptive capacity prevents the expansion of cultivated area. As a result, searching for resistant genes of L. chinensis and uncovering the mechanism of its stress-defence ability are great urgency. Three AOX genes were isolated from L. chinensis by RT-qPCR and RACE, and then sequence analysis was carried out in silico, including analysis of open reading frames, encoded amino acid sequences, protein domains, secondary structures and so on. The open reading frame length of AOX genes were 858, 1 032 and 1 044 bp, which encoded 285, 343 and 347 amino acids, respectively, and then we named the genes as LcAOX1a, LcAOX1b and LcAOX2. Protein homology and phylogenesis analysis revealed that the AOX family protein sequences of L. chinense were highly conserved, especially at the C-terminus, and all the three AOX genes contained “EXXH”, “EEE-Y” iron-binding conserved domains, those may have activities in combination with iron ions. Subcellular localization analysis showed that LcAOX1a protein was localized in other places outside the chloroplast and mitochondria. LcAOX1b protein was localized in chloroplast and mitochondria, while LcAOX2 protein was localized in mitochondrial matrix. The expression patterns of AOX genes were examined by using eight tissues, including stem, leaf, leaf bud, flower bud, calyx, petal, stamens and pistil. The result of RT-qPCR indicated that relative quantity(RQ)of LcAOX1a, LcAOX1b and LcAOX2 genes in floral organs was significantly greater than that in vegetative organs. The RQ value of LcAOX1a and LcAOX1b was the highest in the stamens, especially the LcAOX1a, the expression in the stamens was much higher than other tissues. The LcAOX2 gene has the highest RQ value in the petals. This study cloned three LcAOX genes and performed bioinformatics analysis, subcellular localization analysis and expression patterns analysis to provide a reference for further study of their biological functions.

Cloning and expression analysis of CPP transcription factor family LtTCX2 gene in Liriodendron tulipifera

2020/8/17 12:39:59

CPP(Cystein-rich polycomb-like protein or Tesmin/TOS1-like)protein family are one kind of transcription factors with fewer members and possess a conserved, Cys-rich CRC domains. CPP transcription factor family plays an important role in the plant development of reproductive tissue and cell division. In this study, a CPP-like family gene was cloned by RACE methods from Liriodendron tulipifera named LtTCX2 to explore the function in regulating flower development. The full length of LtTCX2 was 2 866 bp. LtTCX2 contained an open reading frame(ORF)of 2 424 bp, encoding 807 amino acids with two conserved TSO1-like CXC domains by bioinformatics analysis on NCBI site. The molecular weight of protein was 88 699.25 Da, isoelectric point was 5.83 and coefficient of instability was 62.38. LtTCX2 was predicted to be hydrophilic and non-transmembrane nucleoprotein without signal peptide. The amino acid homology sequence and phylogenetic analysis demonstrated that LtTCX2 had high homology with other CPP-like family proteins and was most closely related with NnTCX2 in Nelumbo nucifera and PeTCX2 in Populus euphratica. Real-time quantitative PCR showed that LtTCX2 gene was highly expressed in the leaf and hardly expressed in the sepal and petal. The tissue specific expression order from high to low was that the leaf, flower bud, pistil, stamen, stem, root, petal and sepal. These results suggested that LtTCX2 is a rather conservative gene and provide several help to the phylogenetic evolution of Liriodendron plants from the molecular biology level.

Analysis of ratooning characteristics and quality of distant hybridization buckwheat(Fagopyrum tatari-cymosum)

2021/5/7 15:45:40

‘Guiduoku 75' bred by distant hybridization was selected to investigate the major agronomic traits, yield traits and quality traits in ratooning plants under the 40 cm(I), 20 cm(Ⅱ)and 10 cm(Ⅲ)of stubble height treatment. The results indicated that the plant height, shoot weight, grain number per plant, yield per plant and yield in ratooning plants were significantly altered by stubble height when stubble height was less than 40 cm. The values of plant height, shoot weight, grain number per plant, yield per plant and yield in ratooning plants under treatment I were significant or extremely significant higher than those of treatments Ⅱ and Ⅲ, suggesting that higher stubble height is beneficial for vegetative organ growth and yield formation in ratooning plants. ‘Guiduoku 75' seeds contained four protein components as albumine content > gluten content > prolamine content > globulin content. The total protein content and gluten content were significantly altered by stubble height, but the albumin content, prolamine content, globulin content were not when stubble height was less than 40 cm. The total flavonoid content in ratooning plants of ‘Guiduoku 75' ranged from 3.17% to 3.33%, that was little affected by stubble height. The ratooning plants of ‘Guiduoku 75' under 40 cm of stubble height treatment harvested 1.5 t·hm^-2 seeds that contained 4.06% of albumin content, 3.26% of gluten content, 1.27% of prolamine content, 0.39% of globulin content and 3.33% of total flavonoid content, exhibiting high nutritional value and medicinal value.

Techniques for repeated cutting and transplanting of main stem of virus-free healthy sugarcane seedling propagation

2021/5/7 15:45:40

In order to solve the problems of high production cost, long production cycle and slow propagation speed of sugarcane healthy seedlings, using Guitang 08-120 healthy seedlings as material, after tillers occurred in the original sugarcane virus-free healthy seedlings, the main stem was cut and transplant for two times. The results were as follows: The survival rate of sugarcane healthy seedlings was 48.20% when the main stem was buried and cutting planting seedling was 95.10%; The original seedling planting, the first time cutting transplanting of main stem and the second time cutting transplanting of main stem of the survival rates of transplanting were 97.67%, 96.33% and 96.00%, the number of tillers were 15, 14 and 13, the plant height were 157.67, 127.00 and 123.84 cm, the diameter of stem were 25.52, 25.31 and 25.23 mm, the effective stem number were 87 245, 97 465 and 93 960 bar· hm^-2, the yield were 49 294.5, 52 126.00 and 49 948.50kg·hm^-2, respectively; The first time cutting transplanting of main stem and the second time cutting transplanting of main stem were lower than the original seedling planting, but the tiller number, stem diameter, effective stem number and yield were not significantly different from that of the original seedlings, the results showed that there was no significant differences in sugarcane yield between healthy seedlings of cutting transplanting of main stem and original healthy seedlings; The cost of rapid propagation of one main stem of the original seedling is about 0.47 yuan, which is significantly lower than the cost of conventional propagation of the original seedling. The results of this study provide technical support for reducing the cost and accelerating the breeding speed of sugarcane healthy seedlings.

Analysis of codon usage bias in the chloroplast genome of Dalbergia odorifera

2021/5/7 15:45:41

To comprehend the codon usage pattern of chloroplast genome in Dalbergia odorifera, the 52 coding DNA sequences were analyzed to obtain to the results of neutrality plot, ENC-plot and PR2-plot analysis using Codon W 1.4.2 and online software CUSP in the present study. The results were as follows: The GC content in the three positions of codons from the chloroplast genome of D. odorifera was GC1(46.01%)>GC2(38.98%)>GC3(27.80%)successively. The range of effective number codon was from 37.66 to 54.43, and there were 37 genes when ENC value was greater than 45, and there were 29 genes when RSCU value was greater than 1, including 16 genes ending with U and 12 genes ending with A. These suggest that the codons prefer ends with A and U, and have a weak bias. The neutrality plot showed that there was no significant correlation between GC3 and GC12, the correlation coefficient was 0.250, and the regression coefficient was 0.394; ENC-plot analysis revealed that there were 38 genes which ENC ratio located beyond the section from -0.05 to 0.05; PR2-plot analysis showed that U>A and G>C in the base usage frequency. These all illustrated that the codon usage bias in the chloroplast genome of D. odorifera was mainly affected by mutation; 19 codons were identified as the optimal codon. The present study could be useful in the chloroplast genetic engineering and genetic diversity analysis of D. odorifera.

Growth characteristics of hybrids between Eucalyptus urophylla and Section Exsertaria species

2021/5/7 15:45:41

Deciding crossing parental species will determine characteristics of eucalypt hybrids, but the growth performance of hybrids from different parents is hard to predict. Understanding the growth features of hybrids between important eucalypt species in South China will greatly improve hybrid breeding accuracy. We studied the growth characteristics of hybrids between Eucalyptus urophylla and four Section Exsertaria species/subspecies(E. brassiana, E. tereticornis, E. camaldulensis var. obtusa and E. camaldulensis subsp. simulata)with E. urophylla × E. grandis(UG)hybrids serving as a control, of which the male parents were polymix of 10 plus trees were studied. The results were as follows: Stem volume growth in E. urophylla × Section Exsertaria species(UES)hybrids were significantly lower than UG. And there were significant differences between the UES hybrids(P<0.05), among which E. urophylla × E. camaldulensis subsp. simulata demonstrated significant advantages. DBH and Ht of UES were significantly lower than those of UG, but with height-diameter ratios were significantly higher than UG's(P<0.05), with 5-year-old values approximately 150 and 130, respectively. UES had high and uniform preserve rates, and no significant differences due to either species or family(within species)level. Their 5-year-old values were 84.4%-89.6%. The UES's coefficient of variation(CV)of 5-year-old stem volume had an average of 64%, yet significantly differed between species was higher than that of the UG's. UES was significantly different from UG on volume growth and tree shape, and the significant differences between UES species and families could provide substantial diversity to eucalypt genetic improvement.

Comparison of expression abundance of genes related to rubber biosynthesis regulation in Hevea brasiliensis

2021/5/7 15:45:41
Protein is one of the basic components of life system and the executor of most biological functions. Protein abundance is closely related to its biological function, and its abundance is strictly and precisely regulated by each link in the process of gene expression. Among them, there is a strong correlation between protein abundance and its corresponding mRNA abundance, about 40% of the difference in protein abundance can be explained by mRNA abundance. Jasmonic acid signaling pathway regulates the biosynthesis of natural rubber in Hevea brasiliensis, but the difference of expression abundance among related genes needs to be elucidated. In the present study, the expression abundance differences of 15 rubber biosynthesis regulatory genes COI1, JAZ1, JAZ2, JAZ3, MYC1, MYC2, MYC3, MYC4, MYC5, GAPDH, HMGR1, SRPP, REF, HRT1, HRT2, and 2 common internal reference genes 18S, ACTIN1 in 10 rubber tree germplasms latex following tapping them with S/2D d3 tapping system were compared. The expression abundance of ACTIN1 in each sample is set to 1, and the expression abundance of other genes in the sample is calculated according to the standard. The results were as follows: The transcriptional abundance of different genes in the same individual was significantly different, and the abundance order of the same gene set was different in different individuals; The transcription abundance of the same gene was significantly different in different individuals, the maximum abundance of the 16 genes were 9.43, 6.04, 10.02, 12.29, 18.82, 9.22, 38.46, 112.83, 121.36, 15.34, 19.09, 13.54, 10.05, 19.80, 24.83, 11.82 times of the lowest abundance, and the coefficient of variation were 73.05%, 55.19%, 69.09%, 67.37%, 66.59%, 53.87%, 83.25%, 122.02%, 166.34%, 59.89%, 70.59%, 75.67%, 74.20%, 68.34%, 84.23%, 78.59%, respectively; Overall, at the population level, the transcription abundance of the 16 genes from high to low was 18S>SRPP>HMGR1>REF>MYC2/HRT1>COI1>MYC1/MYC4>GAPDH/JAZ1/MYC5>JAZ2>HRT2/MYC3/JAZ3, correspondingly, the average abundance were 28 382.26, 43.64, 11.39, 7.16, 5.47, 5.10, 1.07, 0.75, 0.74, 0.45, 0.42, 0.33, 0.12, 0.06, 0.06, 0.04 times than that of ACTIN1, respectively. It is worth noting that, the abundance of 18S is undoubtedly the highest, and in mRNA, SRPP is the largest, JAZ1 is greater than that of JAZ2 and JAZ3, MYC2 is greater than that of MYC1, MYC3, MYC4 and MYC5, HRT1 is greater than HRT2 at both the individual and population levels. The results showed that, the abundance of structural genes and functional genes is higher than that of regulatory genes. In the analysis of gene relative expression, the target gene and the internal reference gene are usually homogenized, thus masking the real abundance difference between different genes, therefore, in the gene expression analysis, we should pay attention not only to the relative expression of genes, but also to the abundance difference between genes, which is helpful for understanding the function of genes in a more comprehensive way.

Enhancement of Arabidopsis thaliana seed germination speed by NcEXPA8 gene of Neolamarckia cadamba

2021/5/7 15:45:41
In order to explore the molecular function of NcEXPA8 gene, which is highly expressed in the cambium region of N. cadamba, NcEXPA8 gene expression during seed germination of N. cadamba and the effect of its overexpression on seed germination of Arabidopsis thaliana were studied. The seeds of Neolamarckia cadamba, Arabidopsis thaliana wild type(WT, Col-0)and T3 homozygote of A. thaliana overexpressing NcEXPA8 gene were selected as experimental materials. Quantitative real-time polymerase chain reaction(RT-qPCR)was used to analyze the expression of NcEXPA8 gene in different stages during seed germination of Neolamarckia cadamba, and also its expression level and that of endogenous related genes in the germinating seeds of A. thaliana WT and T3 transgenic homozygotes. Furthermore, the germination rates of WT and T3 homozygous seeds of Arabidopsis thaliana were compared at different treatments and different time. The results were as follows: The expression level of NcEXPA8 gene was different among different stages during seed germination of Neolamarckia cadamba, and its expression level was the highest when the seed coat was broken, and then decreased. Compared to wild type of Arabidopsis thaliana, the overexpression of NcEXPA8 gene not only significantly increased seed germination speed, but also increased the sensitivity to GA and reduced the sensitivity to ABA, but did not affect the expression of endogenous related structural genes in A. thaliana. This study preliminarily analyzes the function of NcEXPA8 gene in seed germination, but its final determination still need to be verified in Neolamarckia cadamba.

Effects of transient infection of FmJAZ1 gene on JA pathway related gene expression in Fraxinus mandshurica

2021/5/7 15:45:41
In this paper, the function of FmJAZ1, a member of the Jasmonates(JAZs), and the influence of its upstream and downstream genes were preliminarily analyzed. Firstly, the FmJAZ1-pROK2-GUS overexpression vector was constructed by using the method of seamless cloning, and then the vector was transferred into Agrobacterium by triparental hybridization. Then, Agrobacterium was used to conduct instantaneous infection on the tissue culture seedlings of Fraxinus mandshurica, and the infected seedlings with FmJAZ1 overexpression were obtained. Finally, the infected seedlings were treated with jasmonic acid synthetic pathway inhibitor(DIECA)at 36 h after infection. Samples were taken at 0, 1, 3, 6, 18, 21, 24 h, respectively. The expressions of FmJAZ1, FmJAZ2, GL1, EIN3 and MYC2 were analyzed by fluorescence quantitative PCR. After Agrobacterium instantaneously infected the tissue culture seedlings of Fraxinus mandshurica, FmJAZ1 expression increased significantly, which was 3.2 times as much as that of no-load infection. After treatment with DIECA, the relative expression level of FmJAZ1 fluctuated significantly at the initial, but was stable after 18 h, which proved that FmJAZ1 was acting on JA pathway. The expressions of JAZ2 and GL1 are down-regulated at 1 h, while the others were slightly up-regulated, and later the expressions of all the genes were up-regulated. Overexpression of FmJAZ1 after instantaneous infection of F. mandshurica indicated that transient infection was effective. After DIECA treatment, FmJAZ1 expression was significantly down-regulated, indicating that the synthesis of FmJAZ1 was regulated by JA. In Fraxinus mandshurica, FmJAZ1 inhibited the expression of transcription factors GL1, EIN3, MYC2, and FmJAZ2, and the synthesis of FmJAZ2 was also regulated by JA. JAZs not only regulates the key proteins of JA pathway, but also participates in the regulation of other signaling pathways, and finally regulates the growth and development of plants and their response to stress through the expression of JA in vivo and the synergistic expression of other related signaling molecules.

Analysis of genetic relationship and construction of fingerprints in Bougainvillea based on ISSR molecular marker

2021/3/8 10:07:44
In this study, ISSR-PCR reaction system was optimized from DNA template concentration, primer concentration, annealing temperature and cycle times, a total of 11 ISSR markers were used to amplify DNA samples of the 131 Bougainvillea cultivars, and the ISSR amplicons were detected based on a agarose gel electrophoresis method. Cultivars genetic diversity was analysed, their genetic distances were calculated, and the clustering analysis and fingerprint construction were performed for all the cultivars. The results were as follows: DNA template concentration was 0.5 ng·μL^-1, primer concentration was 0.5 μmol·L^-1, and the optimal annealing temperature of primers UBC813, UBC814, UBC815, UBC823, UBC824, UBC835, UBC840, UBC841, UBC843, UBC844 and UBC876 were 52.3, 55.9, 54.3, 54.3, 53.6, 56.2, 51.9, 54.4, 54, 50 ℃, respectively, the number of rings was 32. A total of 161 bands were generated collectively by the 11 ISSR primers, and the 156 bands were polymorphic, and the polymorphic ratio was 96.89%. Allele number, effective allele number, Nei's gene diversity index and Shannon's information index ranged from 1.86 to 2.00, 1.33 to 1.68, 0.21 to 0.39 and 0.34 to 0.57, with an average of 1.969, 1.478, 0.294 and 0.447 per primer, respectively. Primer UBC841 had the highest identification rate(80.92%), by which 106 cultivars were identified. A total of 131 cultivars were completely identified and their molecular fingerprints were constructed based on combination of UBC841 and UBC876. The genetic distance between 131 cultivars ranged from 0.00 to 0.60, with an average value of 0.365, and genetic diversity of 131 Bougainvillea cultivars was low, which were divided into six groups at 0.58 genetic distance. Clustering analysis indicates that majority of the cultivars within a species tend to fall in the same cluster, but some cultivars of the same species were grouped in different clusters or sub-clusters and certain cultivars from different species grouped in the same cluster. Genetic diversity of Bougainvillea germplasm resources was revealed accurately, the ISSR-based fingerprints of Bougainvillea cultivar provide reliable technique for cultivar registration and intellectual property protection as well as cultivar clarification in production practices.

Condon preference of chloroplast genome of Drynaria

2021/3/8 10:07:44
The codon usage patterns of the chloroplast genes were related to the expression efficiency of genes. In order to improve the development of the chloroplast genetic engineering of medicinal plant and to promote the quality of medicinal plant Drynaria roosii, the optimal codons in chloroplast genome were identified through analyzing the factors shaping of codon bias of D. delavayi, D. quercifolia and D. roosii using CodonW, CUSP and SPSS softwares. The results were as follows: The effective number of codons(ENC)of the chloroplast gene ranged from 40.10 to 61, from 40.33 to 61, and from 40.15 to 61 in D. delavayi, D. quercifolia, and D. roosii, respectively, which suggested the codon bias was weak among the three species; The ENC of gene ndhE, rpl22, rpl14, rpl20, ccsA, rps4 and rpl16 showed significant difference among species, which suggested the codon bias of some genes was various in closely species; The distribution of ENC frequency ratio of most coding gene ranged from -0.1 to 0.1, suggesting the codon usage bias was produced by gene mutation in Drynaria species. In addition, 12 codons were identified as optimal codons in D. delavayi, ten in D. quercifolia, and three in D. roosii. The codons UCU, ACU, GCU, CAA, AAA and GAU were the optimal codons in D. delavayi and D. quercifolia, the codons UUA and AUU were the optimal codons in D. quercifolia and D. roosii, while did not in D. delavayi and D.roosii. Our results will be useful for the codon preference analysis of ferns and the chloroplast genetic engineering of medicinal plant.

Analysis of codon usage pattern of Eucommia ulmoides

2021/3/8 10:07:44
To comprehend the codon usage pattern of genes in Eucommia ulmoides, taking the E. ulmoides genome codons as the research object, the 320 coding DNA sequences was analyzed to obtain the results of relative synonymous codon usage(RSCU), ENC-GC3s analysis of codon ENC values of coding genes and PR2-plot bias analysis of the codon base usage frequency of the coding genes. By using CUSP and Codon Usage Database software to compare the GC contents and codon occurrence frequency of E. ulmoides with those of Arabidopsis thaliana, Nicotiana tabacum, Escherichia coli and Saccharomyces cerevisiae. The results were as follows: The number of RSCU value greater than 1 had 30 codons, including 18 genes with ending G/C and 12 genes ending with A/U, and these suggested that the codons preferred ends with G/C, and had a strong bias; The number of effective codons(ENC)ranges from 30 to 60, and the codons within this range were far away from the standard curve and their ENC values were smaller and had a higher preference; PR2-plot analysis showed that G>C,U>A, in the base usage frequency; Analysis of GC contents of Eucommia ulmoides and representative species showed that the E. ulmoides GC12, GC3 and average GC contents were higher than those of representative species; The analysis of the codon usage frequency of E. ulmoides and representative species showed that the codon preferences of E. ulmoides and Nicotiana tabacum, Saccharomyces cerevisiae were close, and the codon preferences of Eucommia ulmoides, Arabidopsis thaliana and Escherichia coli were quite different. Eucommia ulmoides is a precious Chinese medicinal material unique in China. Analysis of the codon usage pattern and studies of its codon preference will provide a theoretical basis for the improvement and expression of foreign genes in E. ulmoides plant genetic engineering.

Induction and proliferation of embryogenic callus on improved varieties of fast-growing Pinus elliottii

2021/3/8 10:07:44
In order to make the rapid and large-scale propagation of fast-growing varieties of Pinus elliottii, the induction and proliferation of embryogenic callus were studied. The immature zygotic embryos(with endosperm)of ten P. elliottii families were used to systematically study the effects of different factors on the induction efficiency of embryogenic callus, including genotype, the developmental stage of zygotic embryos, basic medium, types and concentrations of plant growth regulator(PGR). At the same time, the proliferation conditions of embryogenic callus were discussed. The results were as follows: The genotype, the developmental stage of zygotic embryos, basic medium and types and concentrations of plant growth regulator(PGR)all had different degrees of influence on the induction of embryogenic callus. Embryogenic callus were induced in all ten genotypes, of which the induction rate of Genotype 2 was the highest, reaching 25.37%; In the stage of zygotic embryo development, the induction rate of cones at the multiple embryo stage was the highest; Among the four basic media, the induction effect on DCR basic medium was the best; The induction rate of 2,4-D 2.0 mg·L^-1+KT 2.5 mg·L^-1 was the highest, reached 27.78%. The proliferation of embryogenic callus for P. elliottii was achieved on DCR medium, supplemented with 2,4-D(0.5 mg·L^-1), KT(1.0 mg·L^-1), CH(500 mg·L^-1)and Gln(300 mg·L^-1). This study laid a foundation for further developing mature embryo induction and plant regeneration of fast-growing varieties of P. elliottii.

Screening for in vitro rapid propagation conditions of Adansonia digitata

2021/3/8 10:07:44
The purpose of this paper was to establish the tissue culture regeneration system by using the seeds of Adansonia digitata. After treated with a suitable pretreatment and seeds sterilizing method, the initial medium was inoculated to obtain a sterile material. After initiation culture, the multiple shoot were induced in the enrichment medium, and then cut into individual plants for rooting induction, and finally established a tissue culture and rapid propagation system. The results were as follows: The optimal disinfection treatment combination of explants was 75% alcohol treatment for 3 min + 0.1% HgCl₂ treatment for 15 min, the contamination rate of explants was 21.7%; The germination rate was 46.0% after pretreatment with 95% concentrated sulfuric acid for 12 h; The A. digitata seeds were pretreated and sterilized and inoculated onto WPM start-up medium for 48 h, and the germination rate was 74.0%; After the initial culture, axillary buds of the explants were cut into 2-3 cm with a section of young stems and inoculated into the enrichment medium and cultured for 60 d for multiple shoots induction, finally, the best value-added medium was selected as WPM+2.0 mg·L^-1 ZT+ 0.2 mg·L^-1IBA, and the subculture cycle was 60 d with the increment coefficient as high as 3.1; The best rooting medium was WPM + 5.0 mg·L^-1 IBA, and rooting was induced for 60 d with a rooting rate of 61.3%, the average number of roots was 1.9 and the average root length was 8.9 cm. Adding the right amount of IBA can improve the foam root phenomenon and improve the transplant survival rate. After seedling adaptation, the tissue culture seedlings of A. digitata were transplanted to mixed matrix with coconut:vegetable garden soil:perlite(volume ratio 1:1:1), and the survival rate reached above 60.0%. This paper preliminarily established the in vitro rapid propagation technology system of A. digitata, and can provide theoretical and technical supports for the rapid propagation of A. digitata and the preservation of germplasm resources.

In vitro regeneration of Osmunda mildei

2021/3/8 10:07:44
Osmunda mildea is a recently formed natural hybrid species with a narrow distribution area. The number of the population is very small, and its sexual reproduction is extremely low. Using spores as explants, we investigated the effects of basic media(MS, 3/4 MS, 1/2 MS and 1/4 MS)and different plant growth regulators(PGRs)on spores germination, prothallium proliferation and sporophyte formation. The results were as follows: Extremely few fertile spores of O. mildea could germinate and develop into prothallus in effect of plant growth regulators; The optimal PGRs combinations of prothallus growth and proliferation were 1/2MS +1.0 mg·L^-1 6-BA+0.05 mg·L^-1 NAA and 1/2MS +1.0 mg·L^-1 6-BA+0.5 mg·L^-1 IBA; In the 1/4MS with no PGR medium, prothallus were well developed with the proliferation, and the young sporophyte seedling formatted, the formation of scale was 83.49%; The best rooting PGR concentration for young sporophyte was MS+0.5 mg·L^-1 IBA; In loess:river sand:humus=1:1:3 cultivation matrix was the best, the survival rate was more than 98%, and the seedlings thrived. This study is great significance to the artificial breeding and conservation of O. mildea, and also provides reference to the artificial conservation of other endangered fern species.

Analysis of codon usage bias in the chloroplast genome of alfalfa(Medicago sativa)

2021/12/30 15:22:45
To character the codon usage patterns of chloroplast genome in alfalfa, forty-nine coding DNA sequences were analyzed using CodonW, CUSP, CHIPS and SPSS softwares. The results were as follows:(1)The average GC content at the 3rd codon position was 26.44% in the alfalfa chloroplast genome. The effective codon number(ENC)ranged from 40.6 to 51.41, indicating that codon bias was weak in the alfalfa chloroplast genome.(2)Thirty codons showed relative synonymous codon usage(RSCU)value greater than 1, including 29 codons ending with A and U, which suggested that the third codon position preferred A or U.(3)Neutrality analysis showed no significant correlation was observed between GC3 and GC12, suggesting natural selection mainly contributed to the codon usage bias. ENC-plot analysis showed that there were some genes with low ENC values lying below or near the expected curve, which indicated that mutation also affected the formation of codon usage bias. In addition, 17 codons were identified as optimal codons in alfalfa. We concluded that the codon usage bias of alfalfa is formed under effect of natural selection and mutational bias. This study will lay a good foundation for development of chloroplast genetic engineering and molecular breeding of alfalfa.

Codon bias of transcriptomic genes in Cinnamomum camphora

2021/12/30 15:22:45
In order to understand the genes codon bias of transcriptome in Cinnamomum camphora, all transcriptome data of C. camphora were downloaded, screened against their quality. The screened data were assembled and annotated in gene structure after low-quality reads were deleted. Then 37 Mb gene sequences initiated with AUG were extracted by perl script with 34 931 genes. The codon bias was analyzed using CodonW software. The results were as follows: GC content ranged from 0.273 to 0.742 with the average of 0.452; ENC ranged from 26.29 to 61.00 with the average of 52.76; CAI ranged from 0.064 to 0.401 with the average of 0.199; There were 27 genes whose RSCU were greater than 1, of which 22 genes ended with U or A; Neutral plot analysis indicated that a great many genes were not on the line and ENC-plot showed the similar results. The above results elucidate that the gene codon bias of transcriptome was weak, and most ended with U or A; Selection played more important role than mutation in codon bias; Finally, GUU, CAG, GAA, UCU, GCU and GGU were taken as the optimal codons. This will provide a solid foundation for improving traits of C. camphora via the codon revision of the targeted genes.

Rapid propagation technology for eggplant rootstock (Solanums torvum)by micro-cuttage

2021/12/30 15:22:45
Grafting culture is an important measure to prevent soil-borne diseases and increase yield of solanaceous fruit vegetable. Solanum torvum, a wild species of eggplant, has become a common good rootstock of eggplant and tomato because of its strong comprehensive resistance. However, the large-scale application of the seeds in industrial seedling is limited due to the lower germination rate, germination potential and germination index, and the longer seedling age.Therefore, it is urgent to develop other methods and corresponding technical system to improve the seedling efficiency of S. torvum and reduce the seedling cost. In order to optimize the micro-cuttage technique for S. torvum, the stem segment of the first sterile seedling was used as the explants, and different concentrations of plant growth regulators were added to the culture media to compare the effects of different concentrations of plant growth regulators on the micro-cuttage propagation of S. torvum. The results were as follows:(1)There were significant differences in the effect of axillary bud induction, subculture and rooting culture in different media. The optimum medium for primary bud induction was MS+ KT 0.5 mg·L^-1+ IBA 0.1 mg·L^-1, and the budding rate reached 90%.(2)The optimal concentration of plant growth regulators proliferation culture were determined with MS+IBA 0.4 mg·L^-1, in which the average shoot number reached 6.11 per original explant, and the plants grew strong.(3)The root culture of 1/2MS+IBA 0.2 mg·L^-1 was the best, with the number of primary root 4.56, root length 125.80 mm, root diameter 0.50 mm, and numerous fibrous roots. The adoption of micro-cuttings in vitro can realize rapid and mass propagation of S. torvum seedlings. The use of micro-cuttings in the test tube can increase the proliferation coefficient and achieve rapid and large-scale reproduction of S. torvum seedlings.

Optimized study for plantlet inducing from cambium stem cell of young stem based on response surface design in Actinidia chinensis

2021/12/30 15:22:45
In order to reduce the contamination rate and improve the propagation efficiency during the Aitinidia chinensis tissue rapid propagation, the young stem segments of Aitinidia chinensis were used as explants, using the two-step culture method. The response surface design software had been used to optimize the condition or influencing factor for callus inducing and its differentiation in this experiment. The condition was designed at three levels for the concentration of NAA and 6-BA, and hypotonic treatment time, respectively. Meanwhile, the origin of callus and the way of seedling formation were determined by tissue section. The results were as follows:(1)The periderm of young stem was removed using tweezers under aseptic conditions can reduce the contamination rate, and the stems without periderm were treated with 200-400 mg·L^-1 PVP to prevent stem browning.(2)The optimum condition for callus induction was 28.3 h for hypotonic treatment time, NAA and 6-BA concentration was 4.45 mg·L^-1 and 0.28 mg·L^-1, respectively. On the other hand, the optimum condition for callus differentiation or seedling formation was 26.4 h for hypotonic treatment time, NAA and 6-BA concentration was 4.84 mg·L^-1 and 0.42 mg·L^-1, respectively. That suggested that the formation of callus induction required a longer hypotonic treatment time and a higher content of auxin, and a higher content of auxin and kinetin and shorter hypotonic treatment time was required for plantlets inducing.(3)Furthermore, the slice observation during culture indicated that the inducing callus would be derived from division of cambium stem cell and the plantlets would be originated from embryoid development. To sum up, lower contaminated rate, higher propagation coefficient and somatic embryogenesis which had been obtained in this experiment would be based the foundation for the stable propagation system for kiwi fruit industrial culture.

Creating new tobacco germplasm with herbicide- resistance based on oligonucleotide-mediated mutagenesis(OMM)technology

2022/9/30 15:09:08
To further optimize the germplasm of tobacco(Nicotiana tabacum)‘K326', oligonucleotide-mediated mutagenesis(OMM)technique was used in this study. The ALS gene in tobacco ‘K326' was cloned by using the ALS gene sequence reported by NCBI based on the insensitivity and resistance of tobacco to the chlorsulfuron herbicide after the mutation of acetolactate synthase(ALS), the first key enzyme in the branched-chain amino acid synthesis pathway in plants. According to ALS gene sequence, RNA/DNA chimeras were designed for site-directed mutagenesis and introduced into tobacco ‘K326' to create a new tobacco germplasm resistant to chlorsulfuron herbicide. The results were as follows:(1)‘K326'had two ALS genes, namely ALS SuRA and ALS SuRB, with the sizes of 2 004 bp and 2 010 bp, respectively.(2)The chimeras of Chl-588 and Chl-1719 at site 588(Pro site)and site 1719(Try site)of ALS gene were designed according to the conserved sites, ALS SuRA 588 proline site and ALS SuRB 1719 tryptophan site.(3)The two fragments were successfully introduced into tobacco callus by gene gun, and the callus were successively differentiated and rooted by resistant bud, and a total of 22 chlorsulfuron-resistant plants were obtained.(4)The activity of ALS enzyme in resistant plants showed that eight resistant tobacco plants had strong activity, and in the further antagonistic plants, conservative amplification and sequencing of cross-mutation loci resulted in site-directed mutations in two lines(line f11 and line b18)at loci 588 and 1719, respectively. In conclusion, this study provides an ideal parental material for cultivating new germplasm of resistant tobacco while obtaining the new germplasm of ‘K326' resistant to chlorsulfuron.

Identification and bioinformatics analysis of WD40 gene family of Meconopsis horridula

2022/9/30 15:09:08
WD40, as a transcription factor family, is widely involved in regulating plant growth, development, secondary metabolite accumulation and environmental adaptation. In order to explore the role of the WD40 family in the growth, development, secondary metabolite accumulation and tolerance, the WD40 genes of Meconopsis horridula were identified in this study based on the full-length transcriptome sequencing data and conducted bioinformatics analysis of these genes and their encoded proteins. The results were as follows:(1)A total of 19 WD40 genes were identified, and all the proteins included typical WD40 domain, the amino acid numbers and molecular weight of the protein encoded by WD40 genes were 109-758 aa and within 11 830-84 130 Da, respectively, and most of the proteins localized in the nucleus, and all proteins belonged to hydrophilic protein;(2)Phylogenetic tree analysis showed that WD40 proteins of Meconopsis horridula were closely to Papaver somniferum and Macleaya cordata;(3)The promoter region of WD40 gene contained different amounts of hormones or stress-response elements, suggesting that this family genes may be involved in the regulation of various biological processes, such as growth, development and secondary metabolite accumulation;(4)The tertiary structure of WD40 proteins showed that these proteins evolved in different degrees during the evolutionary process. These results can provide a preliminary basis for further research on the specific mechanism of WD40 gene family in response to stress and secondary metabolite accumulation.

Genome-wide identification and expression of millet PAL gene family

2022/9/30 15:09:08
Phenylalanine ammonia-lyase(PAL)involves in the process of phenylpropane metabolism, and PAL gene family plays an important role in the process of plant resistance through regulating the synthesis of plant antibiotics. In order to clarify the expression and the comprehensive analysis of the millet PAL gene family under adversity stress, bioinformatics methods were used to identify and analyze the PAL gene family in millet. The results were as follows: Millet had 11 PAL genes, which were unevenly distributed in different chromosomes. All the PAL gene family contained conserved PAL domains with one to six exons. Phylogenetic analyses showed the PAL gene could be divided into three subfamilies, except that SiPAL7 evolved into one by itself. PAL gene contained cis-acting elements that responded to hormones, adversity stress and other factors, indicating that PAL gene was widely involved in different biological regulation processes. RT-qPCR results showed that millet gene family were mainly inducible expressions, and the expression patterns of PAL gene changed obviously under different light conditions, and different genes had different response patterns, indicating that the millet PAL gene family played an important role in the light regulation response. The PAL gene of millet is highly conserved, responds to different abiotic stresses and has expression specificity. The results suggest that PAL genes may play important roles in the regulation of development and stress responses.

Genome sizes and characteristics of cliff plants Opisthopappus taihangensis and O. longilobus on Taihang Mountains

2022/9/30 15:09:08
Opisthopappus taihangensis and O. longilobus, being perennial cliff herbs and endemic to Taihang Mountains, are important wild germplasm resources of Compositae, and have high economic and ecological values. To determine the appropriate sequencing strategy for the whole genome of O. taihangensis and O. longilobus, in this study, the genome sizes, heterozygosity, GC content, and repeatability were analyzed through the flow cytometry and high-throughput sequencing methods. The results were as follows:(1)Using maize of known genome size as controls, the genome size of O. taihangensis was approximately 2.1 Gb, while that of O. longilobus was approximately 2.4 Gb.(2)For O. taihangensis, the revised genome size was 3.13 Gb, and the repetitive sequences proportion, heterozygosity and GC content in the whole genome were estimated to be 84.35%, 0.99% and 36.56% respectively. Within O. longilobus, the revised genome size, the repetitive sequences proportion, heterozygosity and GC content were 3.18 Gb, 83.83%, 1.17% and 36.62% respectively.(3)The initial depth and content distribution of GC appeared abnormal after initial assembly, which might be related to the relatively high heterozygous rate of the two species. Above all, the whole genomes of O. taihangensis and O. longilobus were both large and complex genomes with high heterozygosity and repetitiveness. Therefore, it suggests that the use of Illumina + PacBio sequencing assembly strategy for the whole genome sequencing analysis of two Opisthopappus species in the future.

Cloning and fuction analysis of RcCATs and RcSODs genes in Tibet Rhodiola crenulata

2022/9/30 15:09:08
In order to explore the role of catalase(CAT)and superoxide dismutase(SOD)in plateau harsh environment adaptation of Tibet Rhodiola crenulata, RcCATs and RcSODs gene family members were analyzed by bioinformatics and qRT-PCR. Spot assay was conducted to study the responses of yeast cells expressing the RcCAT and RcSOD1 genes under abiotic stress. Yeast two-hybrid was conducted to screen interacting proteins from Arabidopsis yeast library respectively by constructing bait vectors of RcCAT and RcSOD1. The results were as follows:(1)There were two CAT genes(RcCAT), three SOD genes(RcSOD), and one Cu/Zn SOD copper chaperone gene(RcCCS). Bioinformatics analysis showed that the above six genes held high sequence identity(66.37%-94.51%)with other homologous species. All genes had no transmembrane domain and held multiple phosphorylation amino acides. Subcellular localization predicted that RcCATs were located in peroxisoma, RcSODs and RcCCS were located in cytoplasm or mitochondria.(2)qRT-PCR analysis showed that RcCATs and RcSODs were constitutively expressed in three organs like root, stem and leaf and held the high expression levels in leaf, and all genes expression levels could also be regulated by low temperature and plant hormones(ABA). RcCAT was significantly up-regulated under cold treatment condition with the highest expression in leaf more than two times higher than root. RcCAT, RcSOD2, RcSOD3 and RcCCS expression patterns were similar under ABA treatment condition.(3)In addition, spot assay showed that the recombinant RcCAT and RcSOD1 yeast cells showed a higher cell viability than the pYES2.0 yeast cells in under cold, hot, NaCl, Na₂CO₃, Co²⁺ and H₂O₂.(4)The pGBKT7-RcCAT and pGBKT7-RcSOD1 bait plasmid without toxicity and auto-activation were constructed to perform yeast two-hybrid screening, then four significant interactional genes with RcCAT were screened, which were AtbHLH121 (AT3G19860), AtCPCK2 (AT2G23070), AtGRP4(AT5G50750)and AtRAPTOR1B (AT3G08850). Total three significant interactional genes with RcSOD1 were screened, which were AtEMB (AT5G11890), AtMBP2 (AT1G52030)and AtRH8 (AT4G00660). These results illustrate that RcCATs and RcSODs play an important role in regulating growth and promoting resistance to environmental stresses in Tibet Rhodiola crenulata, and laid the foundation for in-depth study of the adaptive mechanism of R. crenulata with plateau environment.

Epigenetic diversity of Chinese flowering cabbage revealed by F-MSAP

2022/9/3 13:46:31
In addition to DNA sequence changes, the hybridization Chinese flowering cabbage may also cause epigenetic changes that are independent of DNA sequence. In order to reveal the formation mechanism of epigenetic diversity in Chinese flowering cabbage, the changes on the level and pattern of DNA methylation of 49 Chinese flowering cabbages were tested by F-MSAP. The results were as follows:(1)The detection efficiency of F-MSAP was high, the DNA methylation polymorphism of Chinese flowering cabbage was high, and hybridization could improve the DNA methylation polymorphism.(2)The epigenetic diversity of cabbages was low, the homogenization was serious, and most of the genetic variation was originated from within the species. Selfing increased the epigenetic differences between inbred parents, hybridization increased the epigenetic difference of hybrids.(3)The DNA methylation level of 49 Chinese flowering cabbages was relatively high, the pattern was mainly full methylation. Selfing decreased the DNA methylation level, and hybridization increased the DNA methylation level of inbred hybrids through the change of DNA methylation pattern.(4)The 49 Chinese flowering cabbages were divided into five categories. The results of cluster analysis and principal component analysis were basically consistent. Hybrids tended to be classified according to female parent genetic relationship. This study improves the identification efficiency and accuracy through the analysis of epigenetic diversity of Chinese flowering cabbage, and provides theoretical basis and technical support for further cross-breeding.

Genetic diversity of Pinus taiwanensis var. damingshanensis natural populations by SSR markers

2022/9/3 13:46:31
Pinus taiwanensis var. damingshanensis is a kind of endemic alpine pine of Guangxi and Guizhou, which has high economic and ecological values. Its natural population has not been fully protected and utilized for a long time, which is not conducive to long-term stable development of this species. In order to rationally protect and exploit the natural genetic resources of P. taiwanensis var. damingshanensis, 12 SSR markers were used to study the genetic diversity of three natural populations of P. taiwanensis var. damingshanensis, and to analyze the genetic differentiation and gene flow among populations, so as to provide reference for the protection strategy of this species. The results were as follows:(1)A total of 37 alleles were detected by 12 pairs of primers, and the percentage of polymorphic loci was 100%. For every site, the average number of observed alleles(Na)was 3.08, and the average number of effective alleles(Ne)was 1.68. The number of effective alleles varied greatly among different loci. The average observed heterozygosity(Ho)was 0.35, the average expected heterozygosity(He)was 0.40, and the average polymorphism information content(PIC)was 0.31 for every site.(2)The Shannon's information index of the three populations ranged from 0.48 to 0.65, and the Nei's gene diversity ranged from 0.27 to 0.39. Compared with other related species of pines, the genetic diversity was low. For each population, the average observed heterozygosity was 0.40, and the average expected heterozygosity was 0.33,the average number of effective alleles was 1.58. The genetic differentiation coefficient(Gst)among populations was 0.10. Most of the variation existed in the population and the genetic differentiation level among populations was low. The range of gene flow(Nm)was 2.74, indicating that gene exchange between populations is sufficient of P. taiwanensis var. damingshanensis. This study can provide an important reference for the protection of biodiversity, and lay a foundation for the scientific utilization of P. taiwanensis var. damingshanensis.

EST-SSR marker development and interspecific generality of Albizia odoratissima

2022/9/3 13:46:31
Albizia odoratissima is a unique rare timber tree specie in South China. In order to carry out group genetics research on its germplasm resources, this study designed and developed EST-SSR markers of A. odoratissima based on the transcriptome sequencing results. In addition, A. procera, A. falcataria, Acacia melanoxylon, Erythrophloeum fordii and other related species were selected for analysis of interspecific generality. The results were as follows:(1)Among the 243 pairs of developed primers, 171 pairs could be successfully amplified to the target bands, and the effective amplification rates in Albizia odoratissima, A. procera, A. falcataria, Acacia melanoxylon and Erythrophloeum fordii were 63.79%, 33.75%, 45.68%, 41.56% and 14.81%, respectively, and the polymorphism ratios in them were 23.87%, 12.20%, 9.01%, 3.96% and 2.78%, respectively. There were 18 pairs of primers commonly used among Albizia odoratissima, A. procera, A. falcataria, Acacia melanoxylon and Erythrophloeum fordii.(2)There were 37 SSR polymorphism markers of Albizia odoratissima were obtained, ten polymorphism markers of A. procera and A. falcataria, four polymorphism markers of Acacia melanoxylon, and there was one polymorphism mark of Erythrophloeum fordii.(3)The developed EST-SSR markers can meet the needs of population genetic studies of Albizia odoratissima, and have good transferability and practicability in A. procera and A. falcataria and other related tree species. In conclusion, EST-SSR markers can provide a reliable research tool for genetic diversity evaluation of germplasm resources, fingerprint construction of breeding materials, and population mating system analysis of Albizia odoratissima, A. procera, A. falcataria, Acacia melanoxylon and Erythrophloeum fordii. It is of great significance for the protection and utilization of Albizia odoratissima germplasm resource.

Genomic sequence analysis of four culm variants of Moso bamboo(Phyllostachys edulis)on culm

2022/9/3 13:46:31
As an important and economic bamboo species in China, Moso bamboo has performed lots of variations with long-term cultivation conditions. For an overall understanding of the whole genome of four representative culm variations, Phyllostachys edulis f. holochrysa, P. edulis f. gracilis, P. edulis f. nabeshimana and P. edulis f. huamozhu, re-sequencing was used for high-throughput sequencing to detect its variations by molecular data. The single nucleotide polymorphism(SNP), insertion-deletion(InDel)and structure variation(SV)were detected and annotated, and the mutant genes were compared with the functional databases. The results were as follows: Phyllostachys edulis f. gracilis had the lowest number of mutation sites, that was 11 923, andP. edulis f. huamozhu had the highest number 12 555, of which more than 7 000 mutant genes were annotated. GO annotation classification included 56 functional groups of three functional classification systems: cellular component, molecular function and biological processes. In terms of cell components, there were 2 431 genes related to chlorophyll synthesis. In terms of biological processes, there were 75 genes involved in the synthesis of carotenoids and 80 ones involved in the regulation of anthocyanin synthesis and anthocyanin accumulation in tissues under ultraviolet light. COG classification showed that 369 genes involved in replication, recombination and repair, 291 ones in signal transduction mechanism, and 222 ones in transcription. The metabolic pathways of flavonoids, carotenoids and other substances involved in the mutant genes were analyzed by KEGG database. In-depth study of the regulatory pathways and interpretation of the variation mechanism on culm from the DNA level, can provide a data basis for further exploration of the rich polymorphism and genetic variation of Moso bamboo, and elucidate the genetic basis of gene family and functional genes of different variation types.

Comparative study on chloroplast genomes of three Magnolia species

2022/9/3 13:46:31
In order to investigate the good genes, cultivate the main superior cultivars and discover phylogenetic relationships of Magnolia officinalis, M. officinalis subsp. biloba and M. hypoleuca, we compared the differences among the cp(chloroplast)genomes of three Magnolia species and performed a phylogenetic tree of 14 species. Illumina HiSeq platform was used to sequence and assemble the cp genome of M. hypoleuca. Then the cp genomes of three Magnolia species were annotated by online platform and performed with three Magnolia species cp gene cycles. Moreover, the cp genomes of other 11 Magnolia species were downloaded from the NCBI database and phylogenetic tree of 14 all species cp genomes was constructed based on NJ method. The results were as follows:(1)Clean Reads of M. hypoleuca were 19 791 019, and Q30 was 91.33%. The total length of cp genome of M. hypoleuca was 160 051 bp, its GC content was 39.2%, including 37 tRNA and 8 rRNA.(2)Compared with the cp genome structures of three Magnolia species, three Magnolia species were found to have similar IR, LSC and SSC structures, GC content and tRNA number, but there were differences in the type and number of coding genes, the number and structure of introns and exons.(3)There were six and four more functional gene numbers of M. hypoleuca than the other two Magnolia species, respectively, which indicated that it had stronger viability, and the differential functional genes of three Magnolia species were mainly located in LSC region and IR region, involving large ribosomal subunits, small ribosomal subunits and unknown functional genes groups.(4)Based on NJ phylogenetic tree, M. hypoleuca was closely related to M. officinalis subsp. biloba, next to M. officinalis. In this study, M. hypoleuca has more abundant cp genome structure, composition and variation characteristics, which is the molecular mechanism of its adaptation to low light and low temperature environment in high latitude area. And it will also provide strong guidance for molecular breeding of excellent Magnolia varieties.

Cloning, expression and promoter analysis of LtAGO1 from Liriodendron tulipifera

2022/9/3 13:46:31
To study the morphogenesis mechanism of leaf primordium differentiation, we used Liriodendron tulipifera to clone 2 001 bp upstream region of LtAGO1 CDS as the promoter by RT-PCR and RACE technology and predicted its function, and we also used real-time PCR to investigate expression patterns in Liriodendron, obtained the transgenic Arabidopsis thaliana of ProAGO1∷GUS by resistance screening and DNA dentification, and then monitored phenotype and GUS histochemical staining. The results were as follows:(1) The LtAGO1 gene included an open reading frame for 3 300 bp, encoding 1 100 amino acid, the molecular weight was 122.14 kD and theoretical isoelectric point(pI)was 9.36.(2)Amino acid sequence analysis showed that LtAGO1 consisted of Gly-rich-AGO1 and Piwi conserved domains of AGO family. Phylogenetic trees revealed that LtAGO1 was closed to Cinnamomum micranthum(RWR84608.1)in evolutional relationship.(3)The specific tissue expression analysis demonstrated that the expression order was stamen>floral bud>petal>calyx>leaf>pistil>leaf bud>stem among tissues, the expression order was leaf bud sprouting stage >young leaf stage>senescence stage >mature stage among stages, it was highly expressed in the leaf margin of Liriodendron L., and LtAGO1 gene expression in leaf tooth sinus was higher than that in leaf tooth tip of Liriodendron tulipifera.(4)The transgenic strains leaf polarity of the medio-lateral and proximo-distal axis were absent with serrated leaf margin and double petal flower. It was found that GUS staining was stably detected at the tip of leaf bud of transgenic seedlings, and higher GUS activity was observed at newly differentiated petioles. ProAGO1 promoter drove GUS gene to accumulate specifically in the vascular bundle of Arabidopsis thaliana leaf, flower, pod and stem, and GUS activity intensity order was leaf tip bud> flower>vascular bundle among tissues, which was in accordance with the real-time PCR results. Therefore, the LtAGO1 gene is predominantly expressed in apical meristem and is regulated by various pathways during the development of leaf and flower. The results provide a theoretical basis for further functional research of the LtAGO1 gene in Liriodendron tulipifera and regulation mechanism of leaf shape development.

Cloning and expression analysis of Ml3GT1 in Magnolia liliflora ‘Hongyuanbao'

2022/9/3 13:46:31
UDP-flavonoid 3-O-glucosyltransferase(3GT)is one of the important catalytic enzymes in the anthocyanin biosynthesis pathway. To study the function of 3GT in anthocyanin biosynthesis of Magnolia liliflora, M. liliflora ‘Hongyuanbao' was employed as materials. Primers were designed based on the 3GT sequence obtained from the transcriptome database of M. liliflora ‘Hongyuanbao', and the structural gene Ml3GT1 in anthocyanin biosynthesis pathway was cloned by RT-PCR(reverse transcription-PCR), and its bioinformatics and expression pattern were analyzed. The results were as follows:(1)The cDNA sequence length of Ml3GT1 was 1 863 bp, and the open reading frame was 1 374 bp, encoding 457 amino acid residues. The relative molecular weight of Ml3GT1 was 49.37 kDa, and its isoelectric point was 6.04.(2)The deduced amino acid sequence of Ml3GT1 contains a conserved plant secondary product glycosyltransferase signature sequence(PSPG box).(3)Results of the phylogenetic analysis showed that Ml3GT1 protein was closely relative to 3GT proteins from Freesia hybrida, Petunia &#215; hybrida, and Ipomoea batatas.(4)Results of fluorescence quantitative PCR(qRT-PCR)revealed that Ml3GT1 has spatio-temporal specificity, with the highest expression level in flowers, the lower expression level in young leaves and old leaves, and little expression in roots and stems; With the development of flowers, the expression level of Ml3GT1 gene decreased first, then increased, and showed the highest expression level at the fully-flowering period. These results suggest that Ml3GT1 may be involved in flavonoid 3-O-glycosylation. This study will lay a foundation for the flower and color breeding of Magnolia plants.

Codon usage bias in the chloroplast genome of Ventilago leiocarpa

2022/9/3 13:46:31
To determine the codon usage pattern of chloroplast genome of Ventilago leiocarpa, a total of 50 selected protein-coding sequences were analyzed using Codon W 1.4.2 and online softwares of CUSP and Chips. The results were as follows:(1)There were 29 codons with RSCU > 1, and 28 of them ended with A/U, indicating that synonymous codons in chloroplast genome tend to end with A/U.(2)GC content of codon in chloroplast genome of V. leiocarpa was GC₁(47.38%)> GC₂(39.81%)> GC₃(29.60%), and there were 40 with ENC value greater than 45, which indicates that there is weak bias in chloroplast genome of V. leiocarpa.(3)Neutral mapping analysis and ENC-plot analysis demonstrated that the codon preference of chloroplast genome of V. leiocarpa was affected by both selection and mutation factors.(4)Through the constructed high and low gene expression libraries, 15 optimal codons were finally determined, which were UUG, AUU, GUU, GUA, UCU, CCU, ACU, ACA, GCU, CAA, AAC, GAA, UGU, CGU and GGU. The present study took some basis for the determination of chloroplast genome and genetic diversity analysis of V. leiocarpa.

Bioinformatics of grape CBF4 gene and its response to low temperature and potassium silicate

2022/9/3 13:46:31
In order to explore the structure and expression characteristics of CBF4 gene from grapes, the study analyzed the grape CBF4 gene from the aspects of bioinformatics and low temperature and potassium silicate response. The results were as follows:(1)CBF4 protein was located in the nucleus, there were 5 phosphorylation sites and 14 glycation sites, without signal peptide. It was a hydrophilic, a poor lipid solubility and an extra-cellular protein. The secondary structure was dominated by random coil, with a ratio of 56.88%. The protein contained an AP2/EREBP domain.(2)The multiple sequence alignment and phylogenetic analysis of CBF4 protein showed that wine grapes and American grapes had the highest homology and the closest genetic relationship.(3)Quantitative real-time PCR analysis indicated that the expression level of CBF4 gene in grape leaves was up-regulated after low temperature stress, indicating that CBF4 gene may be involved in the response of grape leaves to low temperature stress. Under low temperature conditions, the CBF4 gene expression was different when potassium silicate was applied, indicating that the response mechanism of this gene to potassium silicate may be different in different grape tissues. These results lay a foundation for further study on the function and mechanism of CBF4 gene in grapes.

Bioinformatics analysis of WRKY gene family in Physcomitrella patens

2022/3/3 16:55:12
As a transcription factor first found in plants, WRKY plays an important role in plant growth and development. However, it has not been studied in Physcomitrella patens. By using the WRKY gene family data(accession number is PF03106)in Pfam database, this paper studied the basic information of WRKY in P. patens, which included physicochemical properties, protein secondary structure prediction, chromosome localization, exon and intron distribution and phylogenetic relationship. The results were as follows:(1)The WRKY gene family in P. patens consisted of 38 members which were divided into two major categories ─Ⅰ and Ⅱ, and the conserved domains of some WRKY genes had mutated.(2)The amino acid length of WRKY protein was 216-775 aa and the relative molecular mass was 24.5-82.8 kDa; Subcellular location showed that the proteins of WRKY gene family were distributed in the nucleus.(3)The secondary structure of WRKY protein was composed of four constituent elements: α-helix, extended strand, β-turn, and random coil; Except for PpWRKY11(α-helix dominated), the proportion of random coil was up to 70%.(4)The phylogenetic relationship with Arabidopsis showed that the number of the WRKY family members and the way of their revolution had changed in the process of plants' evolution, the number of exons of WRKY gene family members were 3-7.(5)The members of WRKY gene were randomly dispersed on 21 chromosomes without forming a gene cluster. This study analyzed the basic structure and properties of the WRKY gene family, and it was found that the WRKY gene family of Physcomitrella patens has evolutionary diversity and unique variability in conserved domains, laying the foundation for subsequent research.

Cloning and expression analysis of BeLEA2 gene from Brachymenium exile

2022/3/3 16:55:12
The purpose of this study was to explore the structural and expression characteristics of LEA2 genes from Brachymenium exile. BeLEA2 gene was firstly isolated and analyzed by polymerase chain reaction(PCR). The results were as follows:(1)Gene structure analysis showed that BeLEA2 gene contained 2 exons and 1 intron and contained an open reading frame(ORF)of 456 bp encoding a protein of 151 amino acids, and its molecular mass was predicted to be 16 515.96 Da.(2)The phylogenetic analysis of LEA2 with other LEA2 in different plants revealed that BeLEA2 from B. exile and LEA2 from Physcomitrella patens belonged to the same branch of evolutionary distance.(3)The promoter sequence of the BeLEA2 gene of 1 072 bp was isolated from Brachymenium exile by high-efficiency hermal asymmetric interlaced polymerase chain reaction(HiTail-PCR)and analyzed by PlantCARE, the results showed that it had TATA-box, CAAT-box, ABRE, MYB, MYC, MYB binding site and other cis-acting elements.(4)Quantitative real-time PCR analysis indicated that BeLEA2 expressed in different stages and tissues of B. exile, and BeLEA2 responded to dehydration stress. These results lay a foundation for further study on the function of LEA2 gene in bryophytes.

Clone and expression analysis of DcSKP1 in Dianthus caryophyllus

2022/3/3 16:55:12
SKP1 gene is the core component of the SCF(Cul1-Rbx1-Skp1-F box)E3 ubiquitin ligase protein complex and is involved in many different biological processes. However, the SKP1 gene of carnation has not been cloned. In this study, a new DcSKP1 gene(the accession number in GenBank was MK931293)was isolated from the flower of Dianthus caryophyllus by using RT-PCR and RACE approaches. The results were as follows:(1)The full length cDNA sequence of DcSKP1 was 962 bp, and a ORF of 567 bp encoded 188 amino acids.(2)Protein sequence alignment showed that DcSKP1 had a highly conserved TPEE motif, Skp1_POZ domain and Skp1 domain, and clustered on a branch with Arabidopsis thaliana Skp1.(3)The DcSKP1 gene expression pattern of Dianthus caryophyllus was studied by qRT-PCR, and it was found that the DcSKP1 gene was expressed in all tissue sites, the expression level in anthers was higher than that in stem and leaf tissues, the expression level was the highest in young anthers, and the expression level decreased with the development of anthers. It is speculated that DcSKP1 gene may play an important role in carnation meiosis in D. caryophllus.

Bioinformatics analysis of GeBP transcription factor gene family in Glycin max

2022/3/3 16:55:12
It has been clarified that GeBP transcription factor regulates the growth and development of plant epidermal hair and participates in the control of plant leaf development. The bioinformatics methods were used to identify the GeBP gene family in the whole Glycin max(soybean)genome, and from physicochemical properties of amino acids, as well as gene structure, physical distribution of chromosomes, phylogenetic tree, and multiple sequence comparison, the functional domain, tissues expression and other basic characteristics of GmGeBP gene family were analyzed. The results were as follows:(1)A total of nine members of GmGeBP transcription factor gene family were identified, of which only two genes contained introns and all had only one intron, indicating that the gene structure of the family members is relatively simple but stable.(2)The molecular weights of GmGeBPs were 39.65-49.24 kD, and the theoretical isoelectric point was 4.65-9.08; these members were basically acidic amino acids, which were hydrophilic and unstable proteins.(3)The chromosome physical distribution showed that nine genes were unevenly distributed on seven chromosomes, two GeBP genes on chromosome 10 and 20 respectively, and one gene on chromosome 3, 5, 13, 15 and 19, respectively.(4)The phylogenetic analysis showed that GeBP members of Glycin max and Arabidopsis thaliana were closely related, clustered into four branches respectively, but far away from Oryza sativa.(5)The analysis of domains showed that all the nine GmGeBP members contained DUF573 domain, which was probably the domain interacting with cis-acting elements of target genes in GeBP transcription factors.(6)By analyzing the expression of GmGeBP transcription factor gene family, we found that the expressions of different genes in different tissues were different, with a certain specificity. The analysis and identification of GmGeBP transcription factor gene family provide the theoretical basis for further studying the molecular role of Glycin max epidermal development.

Cloning and expression analysis of FtF5H gene from tartary buckwheat(Fagopyrum tataricum)

2022/3/3 16:55:13
Ferulate 5-hydroxylase is a key enzyme that regulates the synthesis of S-type lignin. To study the molecular mechanism of ferulate 5-hydroxylase in lignin synthesis pathway of Fagopyrum tataricum, a FtF5H(GenBank accession number: MW455111)gene identified from F. tataricum RNA-seq data was screened and analyzed by bioinformatics methods including the physicochemical properties, signal peptide, transmembrane structure, subcellular localization, hydrophilicity, protein secondary structure and tertiary structure, amino acid structure, as well as phylogenetic tree. In addition, the real-time quantitative PCR(qRT-PCR)was applied to analyze the expression pattern of FtF5H gene in leaves, flowers, stems and husks of thick husk tartary buckwheat and thin husk tartary buckwheat. The results were as follows:(1)FtF5H gene sequence included completed open reading frame with 1 395 bp, coding for 464 amino acids.(2)The FtF5H protein was predicted to have P450 superfamily structure, and FtF5H protein, a non-secretory protein, was a hydrophilic, stable and acidic protein without transmembrane domain.(3)The FtF5H protein secondary structure was predicted to be mainly composed of α-helix and random coil, and its prediction of tertiary structure showed a high similarity with 5ylw.1.A.(4)Phylogenetic analysis showed that FtF5H belonged to the CYP84A subfamily.(5)In addition, qRT-PCR showed that the FtF5H gene was expressed in different parts of the two kinds of tartary buckwheat, and the expression level in the thick husk tartary buckwheat was four times higher than that in the thin husk,with extremely significant differences. The research establish a foundation for further study on the molecular regulation mechanism of lignin synthesis in tartary buckwheat, and has important research significance for the cultivation of new tartary buckwheat varieties.

Screening and transcriptome analysis of EMS-induced cold-tolerant mutants in Hongyang kiwifruit

2023/10/14 20:37:40
The Hongyang kiwifruit (Actinidia chinensis var. chinensis ‘Hongyang')has high economic and nutritional values and good prospects for market development. However, in recent years Hongyang kiwifruit production areas such as Yunnan and Sichuan have been subjected to extreme weather such as inversions on several occasions, and its poor cold resistance has limited its scope for development. In this study, ethyl methanesulfonate(EMS)was used to induce mutants of Hongyang kiwifruit in a tissue culture process, which led to the screening of cold-tolerant mutants and the investigation of their stress response mechanisms through transcriptome analysis. In this study, mutants were induced using EMS induction technology using Hongyang kiwifruit leaves as experimental material in tissue culture(4.4 g·L^-1 MS + 4.5 g·L^-1 agar + 1.5 mg·L^-1 6-BA + 0.1 mg·L^-1 NAA + 15 g·L^-1 sucrose + 0.01-0.10 g·L^-1 EMS)and screened for cold-tolerant mutants under low temperature. Selected cold-tolerant mutants and normal Hongyang kiwifruit tissue culture seedlings were subjected to 4 ℃ 12 h cold stress treatment, while later, transcriptome sequencing analysis was performed. The results were as follows:(1)According to the preliminary phenotypic identification, some of the mutants induced by the 0.06 g·L^-1 EMS were phenotypically resistant to cold;(2)In the GO functional enrichment analysis of transcriptome sequencing data, the most enriched entries were in the biological processes;(3)When using KEGG database analysis, a total of 21 differentially expressed genes(DEGs)were annotated in 15 pathways, and which were all up-regulated. The protein processing pathway(ath04141)in the endoplasmic reticulum had the most DEGs, and sHSF, Hsp70, and NEF in this pathway may be related to the regulation of cold-tolerant mechanisms. The above findings provide a material basis and theoretical rationale for the research and utilization of cold-tolerant germplasm resources of Hongyang kiwifruit.

Sequencing and analysis of full-length transcriptome from Liquidambar formosana leaves in discoloration stage

2023/10/14 20:37:40
Liquidambar formosana is an excellent landscape ecological tree species because its beautiful tree shape and red or orange leaves in autumn. In order to understand the genetic basis of discoloration and secondary metabolism of L. formosana leaves, the mixed samples of L. formosana leaves in five leaf discoloration stages were used for full-length transcriptome sequencing using single-molecule real-time sequencing technology(PacBio platform). The results were as follows:(1)High-quality 41.04 Gb data were obtained by full-length transcriptome sequencing, from which 563 180 full-length non-chimeric sequences were identified, and 27 269 high-quality full-length transcripts were obtained by clustering and de-redundancy. In 27 269 full-length transcripts, 2 035 long-chain non-coding RNA(lncRNA)were predicted, and 14 892 simple sequence repeat(SSR)sites and 1 856 transcription factors were detected.(2)The results of gene annotation showed that a total of 24 857 transcripts were annotated in eight databases such as NR, GO, COG and KEGG, etc., 124 metabolic pathways were obtained in KEGG database, including ribosome, carbon metabolism, amino acid biosynthesis and so on, and 49 and 71 transcripts were involved in flavonoid and chlorophyll metabolism respectively. The above results preliminarily reveal the transcriptome information and functional characteristics of L. formosana leaves during the leaf discoloration stage, and provide basic data for the follow-up study of the molecular mechanism of leaf discoloration, the pathway and regulation of pigment metabolism and synthesis, the cloning of related functional genes and the improvement of leaf color.

Cloning and functional analysis of promoter of TcALDH and TcGLIP genes in Tanacetum cinerariifolium

2023/7/30 11:54:58
Natural pyrethrin is a green botanical insecticide that extracted from the aboveground tissues of pyrethrum(Tanacetum cinerariifolium). Aldehyde dehydrogenase(TcALDH)and GDSL lipase(TcGLIP)are key rate-limiting enzymes involved in pyrethrin biosynthesis pathway in pyrethrum. The promoters of TcALDH and TcGLIP genes were cloned from the genomic DNA of pyrethrum clone ‘W99' in order to investigate the regulatory mechanism of these genes. The regulatory elements, activity, hormone specificity and tissue inducibility of the two promoters were analyzed through bioinformatics analysis, histochemical staining(GUS staining), luciferase reporting, and exogenous hormone treatment. The results were as follows:(1)Using pyrethrum genomic DNA as a template, specific primers were used to clone the pTcALDH and pTcGLIP fragments. The sequence lengths of pTcALDH and pTcGLIP were 2 848 and 1 343 bp, respectively, and the promoter analysis software the PlantCARE predicted that they both contained multiple cis-elements related to stress response and hormone signals.(2)The plant expression vectors fused by pTcALDH and pTcGLIP and luciferase report gene were constructed, and were transformed into tobacco(Nicotiana benthamiana)to analyse hormone inducibility by observing the fluorescence imaging in tobacco leaves. The results demonstrated that the pTcALDH displayed typical hormone inducibility of methyl jasmonate(MeJA)and abscisic acid(ABA), whereas the pTcGLIP showed no response.(3)The tissue culture seedlings of pyrethrum ‘W99' were treated with MeJA and ABA, the expression of TcALDH was up-regulated by ABA within 12 h, and first increased and then decreased under MeJA treatment; the expression of TcGLIP was down-regulated by ABA and MeJA.(4)We constructed the expression vectors of pTcALDH and pTcGLIP fused with GUS reporters and transformed them into tobacco, then the transient transformation of tobacco drived the expression of GUS gene and showed initiating activity. It was found that the pTcALDH expressed in both the glands, glandular hair heads and mesophyll of the leaves, while the pTcGLIP was only expressed in the parenchyma cell. These results indicated that the pTcALDH and pTcGLIP were tissue-specific promoters, and the pTcALDH appeared MeJA-inducible and ABA-inducible characteristics. This study provides a new insight into the regulatory mechanism of TcALDH and TcGLIP genes involved in pyrethrin synthesis.

Cloning and expression analysis of FeR2R3-MYB of anthocyanin synthesis-related genes of common buckwheat

2023/7/30 11:54:58
MYB is a common transcription factors widely involved in the regulation of anthocyanidin biosynthesis. In order to explore the regulatory role of MYB transcription factors in the biosynthesis of common buckwheat anthocyanidins, this study screened and cloned a MYB gene associated with anthocyanin biosynthesis from the transcriptomic data of safflower common buckwheat and white flower common buckwheat, and named it FeR2R3-MYB, GenBank login number was MT151381.1. The sequence was analyzed by bioinformatics analysis and qRT-PCR was used to analyze the expression characteristics of FeR2R3-MYB gene in white flower common buckwheat and safflower common buck wheat. The results were as follows:(1)FeR2R3-MYB gene was 831 bp in total length, encoding 276 amino acids. The relative molecular mass of the protein was 30.95 kD, the theoretical isoelectric point(pI)was 8.73, and the instability index of the protein was 69.64, which belonged to the unstable protein. The total hydrophobic value was -0.679, and the whole peptide chain showed hydrophilic characteristics.(2)FeR2R3-MYB had a typical R2R3-MYB domain and belonged to the R2R3-MYB subfamily.(3)FeR2R3-MYB was closely related to common buckwheat and knotweed, belonging to the same family.(4)The promoter sequence of FeR2R3-MYB contained a total of nine light corresponding elements, 17 transcription factor binding sites, four abiotic corresponding elements and two hormone response elements.(5)Subcellular localization found that FeR2R3-MYB was only expressed in the nucleus.(6)The expression of FeR2R3-MYB gene of safflower common buckwheat was higher than that of white flower common buckwheat in leaves and inflorescences, and it was further speculated that FeR2R3-MYB gene could positively regulate the biosynthesis of common buckwheat anthocyanin. In summary, these results lay a foundation for further deepening the research on the function and expression regulation of FeR2R3-MYB gene in the biosynthetic pathway of common buckwheat anthocyanin.

Effects of genetic improvement on traits of needle and cone of Cunninghamia lanceolata

2023/7/30 11:54:59
To reveal the effects of genetic improvement on leaf and seed traits of major afforestation timber species, and to clarify the variation trend of traits, selected trees(improved population)of the fourth cycle of Chinese fir breeding and phenotypic superior trees and ancient trees(unimproved population)in four provinces and five regions were selected as the research objects. The needle and cone traits of 218 clones were investigated, and the study used variance analysis and multiple comparison methods to study the phenotypic differences of genetic improvement on Chinese fir and different types of Chinese fir. The correlation analysis was used to explored the effect of genetic improvement on the phenotypic traits of Chinese fir needles and cones; the principal component analysis and cluster analysis were used for classification. The results were as follows:(1)The needle length, needle width and seeding rate of the unimproved population were 13.28%, 10.81% and 33.90% smaller than those of the improved population, respectively. Other traits showed that the unimproved population was larger than the improved population, with differences ranging from 10.90% to 27.03%. The coefficients of variation of cone length, cone width and seeding rate of the unimproved population were 9.14%, 12.73% and 15.38% larger than those of the improved population, respectively.(2)Among the four traits of cone length, cone width, bract scale length and bract scale width, only cone length and cone width(0.931), bract scale length and bract scale width(0.622)were extremely significant positive correlation between them in the unimproved population. After genetic improvement, the four traits showed a significant or extremely significant positive correlation in pairs.(3)Cone length and cone width in Sichuan Ya'an(SCYA)were 48.83% and 53.26% larger than those in the improved population, and the hundred-grain weight in Anhui Huangshan(AHHS)was 16.92% larger than those in the improved population.(4)Genetic improvement led to a decrease in the proportion of Chinese fir with loose-stretched cones, and an increase in the proportion of Chinese fir with tightly packed and inverted cones. In summany, this study believes that the genetic improvement of Chinese fir leads to the reduction of cone size, changes the proportion of different needles and cone types, and changes the correlation between needle traits and cone traits, which will be helpful for providing the basis to the evaluation of Chinese fir germplasm resources and future multi-targets breeding.

Optimization of protoplast preparation conditions of Pleurotus giganteus

2023/7/30 11:54:59
In order to obtain the optimal conditions for the protoplast preparation of Pleurotus giganteus, two strains, PG46 and PG79, with different temperature types, were selected as materials to study the effects of five factors(mycelial age, osmotic stabilizer type, lywallzyme concentration, enzymatic hydrolysis duration and enzymatic hydrolysis temperature)based on single factor and orthogonal experimental methods. The results were as follows:(1)In the single factor test, the optimal conditions for the protoplast preparation of P. giganteus were mycelial culture for 5 d, using 2.5% lywallzyme with 0.6 mol·L^-1 mannitol, incubated for 4 h at 32 ℃(PG46)or 27-35 ℃(PG79).(2)Orthogonal experiment verified and optimized the single factor test results. Combination 2(mycelial age 5 d, lywallzyme concentration 2.5%, 0.6 mol·L^-1 mannitol, incubated for 4 h at 32 ℃)could be used as the optimal condition for the protoplast preparation of PG46 and PG79 at the same time, and the protoplast yields were 11.22 × 10⁶ CFU·mL^-1 and 7.28 × 10⁶ CFU·mL^-1, respectively.(3)For F-test, the influence degree of various factor on the protoplast preparation were as follows: mycelial age>lywallzyme concentration>enzymatic hydrolysis temperature>enzymatic hydrolysis duration(PG46), and mycelial age>enzymatic hydrolysis duration>enzymatic hydrolysis temperature>lywallzyme concentration(PG79), respectively. In conclusion, the protoplast preparation conditions of the two P. giganteus strains with different temperature types were basically the same, and the effects of mycelial age on the protoplasts yield of the two strains were the most significant. The results can lay a foundation for further cross-breeding, genetic transformation, whole genome sequencing and promote the molecular genetics development of P. giganteus.

Genetic diversity and population structure of Aegilops tauschii accessions based on ISSR method

2023/7/30 11:54:59
Aegilops tauschii is considered as an important gene source for improving common wheat, which has wide distribution and rich genetic variation. In order to understand genetic diverstity and population structure of A. tauschii from different origins, ISSR markers were used to evaluate genetic diversity and population structure of 56 A. tauschii accessions. The results were as follows:(1)The 170 polymorphic bands were detected by 16 ISSR primers, and polymorphic bands of each ISSR primer ranged from 3 to 18, with an average of 10.63. The variation of polymorphism information content(PIC)ranged from 0.17 to 0.85, with an averaged of 0.67.(2)The comparison among four populations indicated that the population genetic diversity of Central Asia was the highest(H_e=0.225 4, I=0.355 7)and gene flow between populations was relatively low(N_m=1.638 6).(3)The clustering results showed that 56 A. tauschii accessions were divided into two groups at the genetic similarity coefficient 0.67, of which eight accessions from Tajikistan and Turkmenistan were clustered in Group 2. And, the Group 1 including 48 accessions could be further divided into three sub-groups, which indicated that A. tauschii accessions with clustering together have the same origin.(4)Based on population structure analysis, 56 A. tauschii accessions were divided into five populations, of which the V population from Iran in West Asia had relatively consistent genetic background and relatively low degree of hybridization. Furthermore, the Q value analysis of populations showed that the genetic relationship of IV population were relatively complex, producing the most abundant genetic diversity. The results of this study can provide an important reference for analysis of genetic relationship, protection of biodiversity, and lay a foundation for the scientific utilization and evolution research of A. tauschii.

Identification of resistance to root rot and preliminary establishment of its SSR markers in castor bean

2023/7/30 11:54:59
Castor bean root rot is a root disease caused by Fusarium solani, which seriously threatens the production of castor bean. Due to the lack of resistance genes, the breeding for root rot resistance in castor bean is seriously restricted. In order to mine resistant resources and establish resistant molecular markers, the phenotypic and molecular marker identification was performed on the disease resistance of 252 castor bean accessions in this study. The results were as follows:(1)Irrigating roots with the conidia suspension of 1×10⁶ spores·mL^-1 was an effective inoculation method. The 5-grade evaluation method based on the days of wilt after inoculation could be used as the criteria to evaluate the resistant level of accessions objectively.(2)According to the criteria, the resistance of 252 accessions were divided into five grades from high to low, among which Grade 1 was high resistance and Grade 2 was moderate resistance. The number of accessions with different grades from 1 to 5 were 105, 25, 33, 31 and 58, respectively, accounting for 42%, 10%, 13%, 12% and 23%, respectively. A total of 130 resistant accessions were identified, of which 105 were high resistance and 25 were moderate resistance.(3)The proportion of resistant accessions in wild accessions(66%)was much higher than that in cultivated accessions(35%). Among wild accessions from South China, 69% were resistant accessions, and 60% were high resistance accessions. It is strongly suggested that the research and utilization of wild accessions, especially the wild accessions in South China, should be an important direction of resistance breeding in the future.(4)Eight SSR markers associated with the resistance were preliminarily established. Although different resistant accessions carried different markers or marker combinations, most of them carried three to four of the above markers, therefore, they can be used as resistant molecular markers for assisted selection. The results of this study provide an effective method and evaluation criteria for root rot resistance identification, screen out a number of resistance genetic resources urgently needed in breeding, and preliminarily establish the SSR markers available for assisted selection, which lay an important foundation for resistance breeding of castor bean root rot.

Full-length transcriptome information for Tibetan medicine “Zangyinchen” of original plant Comastoma polycladum

2023/7/30 11:54:59
Comastoma polycladum is one of the original plant of Tibetan medicine “Zangyinchen”, which contains abundant medical components. To further know the transcriptome of C. polycladum and enrich its genetic information of gene annotation and metabolic pathway, the Pacbio sequencing platform was used to perform full-length transcriptome sequencing of C. polycladum leaves. The results were as follows:(1)A total of 17 Gb of sequencing data was collected, and 87 814 high-quality full-length transcripts were obtained by clustering and de-redundancy of 795 698 CCS sequences.(2)Comparing with seven databases, 277 451 transcripts were annotated successfully, and in NR database with 39 214 transcripts annotated the most transcripts. A total of 26 396 transcripts were annotated to the KOG database, with 26 subcategories, and a total of 39 104 transcripts with six major pathways and 40 secondary pathways to the KEGG database. A total of 39 102 transcripts were annotated to the GO database, which were divided into three major categories: molecular function, biological process and cellular component.(3)SSR analysis yielded 22 861 SSRs, with single-base repeats being the most abundant. A total of 1 874 transcription factors and 15 166 long non-coding RNAs(LncRNAs)were identified, and the C3H transcription factor family had the most annotated transcripts.(4)A total of 55 transcripts involved in the synthesis of monoterpenes and flavonoids were screened out. These results enrich the transcriptome information of C. polycladum, and provide significant genetic resources for further screening of candidate genes related to the synthesis of medicinal components of C. polycladum.

Identification and evolutionary analysis of cotton DUR3 gene

2023/3/26 12:05:35
Plant DUR3 homologous protein is a high affinity urea transporter which belongs to the family of sodium/solute symporter family, and plays an important role in the active absorption of exogenous urea and redistribution of endogenous urea by plants. The purpose of this study was to clarifystructure and evolution situation of cotton DUR3 gene. Based on bioinformatics methods, DUR3 genes were identified from Gossypium hirsutum and G. raimondii genomic sequences, and then the gene structure, transmembrane domain, motif location, as well as phylogenetic relationship, were systematically analyzed. The results were as follows:(1)Three DUR3 genes were identified from the A and D subgroup chromosomes of upland cotton and G. raimondii genomic sequences. These three cotton DUR3 homologous proteins, like other plant DUR3 homologous proteins, had 15 transmembrane domains and three highly conserved motifs with consistent positions.(2)The gene structure analysis showed that the number of exons of DUR3 genes in dicotyledons was significantly higher than that in monocotyledons, and so were the cotton DUR3 genes.(3)Phylogenetic analysis revealed that the amino acid sequences of different species were classified according to species kinship, and cotton clustered in one branch with dicotyledons.(4)The K_a/Ks values of orthologous and paralogous DUR3 genes were generally more than one, indicating that those genes mainly experienced positive selection among evolution. The results of this study provide a theoretical basis for further research on cotton DUR3 homologous protein.

Identification and analysis of TCP transcription factors in response to low nitrogen fertilizer stress in Solanum tuberosum

2023/3/26 12:05:35
TCP transcription factors are a type of plant-specific transcription factors, which have been implicated in multiple aspects of plant biological processes. To study the role of potato TCP transcription factors in response to low nitrogen fertilizer stress, here, four transcriptome libraries from potato roots and leaves under the conditions of low nitrogen fertilizer(0.05 mmol·L^-1)and sufficient nitrogen fertilizer(7.5 mmol·L^-1)were constructed for transcriptome sequencing, respectively. Meanwhile, the differentially expressed TCP transcription factor were analyzed. The results were as follows:(1)A total of 24 TCP transcription factors were identified in the four transcriptome libraries, and mainly distributed on the second, third and sixth chromosomes.(2)The analysis of domains showed that all of 24 transcription factors contained basic-Helix-Loop-Helix domain.(3)The phylogenetic analysis showed that the TCP proteins of potato and Arabidopsis thaliana were closely related, and clustered into 10 subgroups.(4)Transcriptome sequencing results showed that the expression levels of most potato TCP transcription factors were suppressed by low nitrogen fertilizer stress. Among them, three TCP transcription factors were significantly differentially expressed in roots, while five TCP transcription factors were specifically expressed in leaves.(5)According to the GO functional analysis and the relationship between the potato and the A. thaliana TCP transcription factors, it is predicted that these TCP transcription factors were involved in response of potato to low nitrogen fertilizer stress. The research provides a foundation for further study on the molecular role of transcription factors in response to low nitrogen fertilizer stress in potato and other crops.

Discovery and analysis of volatile oil synthesis related genes in Illicium verum based on transcriptome sequencing

2023/3/26 12:05:35
In order to identify volatile oil synthesis related genes in Illicium verum, transcriptome sequencing and assembly annotation were carried out on the leaves of the superior clone Guijiao 69 and common variety Zhen 01, two varieties with significant difference in volatile oil content. Then GO and KEGG pathways analyses of differentially expressed genes(DEGs)were performed. The results were as follows:(1)A total of 84 182 Unigenes were obtained after transcripts assembly, and 59 161 Unigenes were annotated in NR, NT, Swissprot, KEGG, KOG, GO, and Pfam databases. A total of 30 572 DEGs were obtained after filtering the low abundance genes, and 15 025 up-regulated and 15 547 down-regulated genes were identified in Guijiao 69 compared to the common variety Zhen 01.(2)GO classification results showed that 20 287 DEGs were annotated. In addition, 21 600 DEGs were involved in 133 KEGG pathways. Among them, genes encoding the key enzymes related to volatile oil synthesis such as linalool synthase, myrcene synthase, geranyl geranyl pyrophosphate synthase, cinnamoyl CoA reductase, caffeic acid 3-O-methyltransferase and cinnamyl alcohol dehydrogenase were differentially expressed and enriched in monoterpene biosynthesis pathway, terpene skeleton biosynthesis pathway and phenylpropanoid biosynthesis pathway.(3)Transcription factor analysis showed that the DEGs were distributed in 31 transcription factor families, of which MYB family had the most Unigenes. In this study, transcriptome sequencing was performed to analyze the DEGs superior clones and common varieties of I. verum as well as their related functions and pathways. The candidate genes obtained provide the references for further exploring the synthesis mechanism of characteristic components of volatile oil and molecular breeding of I. verum.

Cloning, prokaryotic expression and subcellular localization of PtCHI gene from Pueraria montana var. thomsonii

2023/3/26 12:05:35
In order to explore the differences in the molecular mechanism of the Pueraria cultivars between isoflavone metabolic enzyme gene PtCHI, and to preliminarily reveal the difference content causes of the isoflavones. The materials of the study were Pueraria montana var. lobata and P. montana var. thomsonii. Puerarin and total flavonoids of P. montana var. lobata and P. montana var. thomsonii were extracted by ethanol, and their contents were measured by high-performance liquid chromatography. Based on the reported CHI gene of Pueraria montana var. lobata, the PtCHI gene from P. montana var. thomsonii was isolated by homologous cloning method, and the protein was expressed in vitro. At the same time, the location of the PtCHI gene was studied in Arabidopsis thaliana protoplasts. The results were as follows:(1)The content of puerarin in P. montana var. lobata was significantly higher than the P. montana var. thomsonii, and the content of total flavonoids was also higher but not significant.(2)The gene PtCHI was successfully isolated from P. montana var. thomsonii. The gene was 742 bp in length, containing a complete ORF frame of 672 bp, encoding 223 amino acids, and had up to 99% homology with P. montana var. lobata.(3)This study found that the expression of CHI gene in P. montana var. thomsonii was stem>root>leaf, P. montana var. lobata was root>stem>leaf. The expression of CHI gene from P. montana var. lobata was significantly higher than P. montana var. thomsonii besides in leaves.(4)Through the online tool prediction analysis, PtCHI was found to be stable hydrophilic protein and the size was 27.8 kD. The secondary and tertiary structures were based on α-helix, with 25 phosphorylation sites, closely relating P. montana var. lobata, Glycine max and Glycyrrhiza uralensis, and were more likely to interact with F3H2, F3H, 4CL4, DFR2 and CHS.(5)At the same time, the protein of PtCHI was successfully induced and isolated in vitro, with a single protein of 27.8 kD.(6)Through the Arabidopsis thaliana protoplasts revealed that PtCHI was mainly located in the chloroplasts. This study is in further analyzing the difference in flavonoids in P. montana var. lobata and P. montana var. thomsonii, as well as laying the foundation for the functional verification of P. montana var. thomsonii PtCHI and the research on the mechanism of isoflavone metabolism.

Development and validity evaluation of Liquidambar formosana EST-SSR primers based on transcriptome sequencing

2023/3/26 12:05:35
Liquidambar formosana is one of the important native tree species in Guangxi, which has high timber, ornamental and medicinal value. This study designs and develops EST-SSR markers of L. formosana based on data from the transcriptome sequencing. Primers were developed, and screened out by PCR amplification and polyacrylamide gel electrophoresis for high polymorphism, and the efficiency was tested by using 30 individuals from a wild L. formosana population to verify the practical application of these SSR primers. The results were as follows:(1)A total of 23 777 SSR loci were obtained by searching SSR Unigenes from transcriptome data. The repeat type of the SSR loci in L. formosana was dominated by mononucleotide repeat type(46.54%). The highest proportion of SSR loci(72.36%)was between 5 and 12 times.(2)A total of 262 pairs of SSR primers were developed, and among them, 139 pairs of effective amplifications with a success rate of 53.1%, and 18 pairs of them that could be used to steadily obtain clear bands were finally identified.(3)The polymorphism detection showed that all sites had a high degree of polymorphism. The number of alleles(Na), effective alleles(Ne), Shannon's diversity index(I), observed heterozygosity(Ho)of the L. formosana population ranged in 2-4, 1.112 8-2.609 6, 0.208 9-1.112 7 and 0.275 9-1.000 0, the average values were 2.333 3, 1.957 4, 0.708 5 and 0.722 6, respectively. In conclusion, the dominant SSR repeat type and repeat motif in L. formosana are basically the same as those in other species, the developed 18 pairs of EST-SSR markers can meet the needs of population genetic studies of L. formosana, which provides abundant primers for the study of genetic diversity of L. formosana. It is of important significance to the protection, development and utilization of L. formosana.

Functional analysis of RcMsc2 gene in castor(Ricinus communis)

2023/3/26 12:05:35
G2/mitotic-specific cyclin-2(Msc2), as a key regulatory protein in response to stress in higher plants, is involved in multiple responses to stresses. In order to explore the function of RcMsc2 gene which was successfully cloned from castor leaf tissue, the structure and potential function of RcMsc2 protein were analyzed by bioinformatics, and the expression characteristics of RcMsc2 gene in tissue and abiotic stress were analyzed by qRT-PCR. The results were as follows:(1)RcMsc2 gene was located in the long arm of Chromosome 5 in castor, and its CDS region was 1 299 bp, encoding 432 amino acids.(2)RcMsc2 protein has the characteristic domain of cyclin family, which was an unstable acidic hydrophilic protein without transmembrane domain and signal peptide, and its relative molecular weight was 49.38 kD.(3)The secondary and tertiary structures of RcMsc2 protein were mainly α-helix and random coil.(4)RcMsc2 protein had the highest sequence homology with CYCB2 protein of Jatropha curcas and Hevea brasiliensis, and was clustered into Group Ⅱ.(5)35S-RcMsc2-GFP fusion protein was localized to the nucleus.(6)RcMsc2 gene was expressed in all tissues of castor, and mainly played a role in roots and stems; abiotic stress analysis showed that RcMsc2 gene could be induced by abscisic acid( ABA ), salt, drought and low temperature treatment, and the response of RcMsc2 gene to low temperature stress was the most sensitive. In summary, this study comprehensively analyzed the structural characteristics, phylogenetic evolution and expression patterns of RcMsc2 gene, and provides a theoretical reference for revealing the function of RcMsc2 gene in castor growth and development and response to cold stress.

Cloning and expression analysis of the SpLEA1 gene of Selaginella pulvinata under drought stress

2023/3/26 12:05:35
Late embryogenesis abundant(LEA)is widely present in organisms and closely related to plant resistence, it can protect plant cells and reduce plant damage under drought stress. Selaginella pulvinata is a fern with the ability to survive drought stress, with a strong recovery ability under drought stress. To investigate the molecular mechanisms and expression characteristics of the SpLEA1 gene in drought-tolerant plants, we used the highly drought-tolerant plant S. pulvinata as experimental material and obtained the cDNA sequence of the SpLEA1 gene by RT-PCR based on the transcriptome sequencing results. The promoter sequence was obtained by the HiTail-PCR technique, and the sequence was analyzed by bioinformatics. qRT-PCR was used to analyze the expression pattern of the SpLEA1 gene under drought stress. The results were as follows:(1)The length of SpLEA1 was 476 bp, the open reading frame(ORF)was 279 bp, and it encoded 92 amino acids. The predicted molecular weight of the protein was 9 491.46 Da, and the isoelectric point was 5.45. The predicted protein structure analysis showed that the protein was hydrophilic. The protein contained ten phosphorylation sites, of which six serines, three tyrosines, and one threonine, respectively, and the predicted secondary structures showed that the protein was mainly composed of α-helix and random coil.(2)The conserved structural domain of the SpLEA1 protein was predicted to be Lea-5, derived from the LEA1 family. Based on the phylogenetic tree and genetic distanced matrix, the SpLEA1 was found to have high homology with Lea-5 protein from Cicer arietinum and Trifolium pratense.(3)Predictive analysis of cis-acting elements in promoter sequenced revealed that the SpLEA1 gene promoter contained five classes of hormone response elements and functional elements related to the drought stress response. The SpLEA1 gene was hypothesized to have multiple functions in the plant body and was closely related to drought stress response mechanisms.(4)SpLEA1 gene expression was up-regulated under natural dehydration treatment and peaked in 12 h. After rehydration treatment for 24 h, expression was significantly down-regulated. In summary, the SpLEA1 gene is likely to be involved in the regulation of drought stress response mechanisms in matted curly cypress. This results provide the reference for further studies on the function of the matted cypress SpLEA1 gene under drought stress and its expression regulation mechanism.

Accumulation rule of main nutrients and isoflavones and their relationships in Pueraria lobata var. thomsonii

2023/12/30 21:36:09
In order to determine the optimal harvesting period and the application value of each part of Pueraria lobata var. thomsonii and to explore the correlation between the main nutrients and the isoflavones, P. lobata var. thomsonii of Tengxian, Guangxi was used as the material, the contents of isoflavones in different parts, the contents of main nutrients and isoflavones in different growth periods were determined by test box method, enzyme gravimetric method, Soxhlet extraction method, high performance liquid chromatography and other physiological and biochemical techniques, and the relationship between the accumulation of various nutrients and isoflavones was analyzed to clarify the accumulation rule of main nutrients and isoflavonoids and their relationships in P. lobata var. thomsonii. The results were as follows:(1)At the mature stage, the content of genistein in different parts of P. lobata var. thomsonii was not significantly different, but the content of isoflavoues aglycone in leaf was significantly lower than that in middle vine, and the contents of total isoflavones, puerarin and daidzein in root apex and vine were significantly higher than those in leaf and tuberon, and the contents of puerarin in root apex and vine were more than 1.00%.(2)The accumulation of starch, polysaccharide, crude protein, and soluble protein reached the maximum in November and December, the accumulation of insoluble dietary fiber reached the minimum in December, the accumulation of soluble dietary fiber was the maximum in August and December, and the accumulation of total isoflavones and puerarin reached the maximum in August and September.(3)The correlation analysis results showed that the accumulation of total isoflavones and puerarin was negatively correlated with the accumulation of starch and polysaccharide; the accumulation of total isoflavones and puerarin was positively correlated with the accumulation of insoluble dietary fiber. Therefore, it can be concluded that the root apex and vine of pueraria powder contain rich total isoflavones, puerarin, daidzein, which has good value for medicinal development; the best harvest time for medicinal using are August and September, and the best harvest time for edible using are November and December; the accumulation of total isoflavones and puerarin is related to starch, polysaccharide and insoluble dietary fiber. This study clarify the accumulation rule and correlation of the main nutrients and total isoflavones in P. lobata var. thomsonii, and provides a reference for the comprehensive development and utilization of P. lobata var. thomsonii and the determination of the harvest time.

Physiological differences of different rice materials in response to salt stress at seedling stage

2023/12/30 21:36:09
In order to investigate physiological characteristics of salt-tolerance rice HD96-1 at seedling stage, indica rice HD96-1(strong salt-tolerant)and 93-11(weak salt-tolerant)were used as experimental materials in this study. Three NaCl concentrations(0, 60, 120 mmol·L^-1)was set to treat 3-leaf rice seedlings for 7 d. The growth, physiological and biochemical indexes of two rice cultivars were measured and analyzed. The results were as follows:(1)The plant height and the pseudostem width of two cultivars decreased, and the root-shoot ratio increased under salt stress; compared with 93-11, the decrease of plant height and pseudostem width of HD96-1 was lower, and the increase of root-shoot ratio was higher under salt stress; in addition, the dry weight of aboveground and root of HD96-1 increased under salt stress, while that of 93-11 decreased.(2)The malonaldehyde(MDA), superoxide anion(O₂^-)and hydrogen peroxide(H₂O₂)contents in rice seedlings increased after salt stress, of which the increase of HD96-1 was lower than that of 93-11.(3)The activities of superoxide dismutase(SOD), peroxidase(POD), catalase(CAT)and ascorbate peroxidase(APX), the contents of ascorbic acid(AsA), glutathione(GSH), proline, soluble sugar and soluble protein in rice seedlings were increased under salt stress, of which the increase of HD96-1 was higher than that of 93-11. In conclusion, the physiological mechanisms of two rice seedlings responding to salt stress are similar, but the difference is that HD96-1 with strong salt-tolerance is stronger antioxidant and osmotic regulation ability than 93-11 in response to salt stress, so that growth and development of HD96-1 are less inhibited than 93-11.

Effects of GABA seed priming on alleviating salt stress and physiological and biochemical changes in Capsicum annuum

2023/12/30 21:36:09
Seed priming is proved to be an efficient way to improve crop salt tolerance, which is a measure of pre-treating seed with various agents while obtaining enhancement to crop performance at growing stage. Gamma amino butyric acid(GABA)is a non-protein amino acid involved in various metabolic processes, and partially protects plants from environmental stress. Enhancing effects of GABA priming on germination characteristics and abiotic stress have been established in several crops. However, the effect and mechanism of GABA seed priming on salt tolerance of pepper(Capsicum annuum)are still unknown. In this study, a hybrid pepper variety of ‘Maoshu 360' Chaotian pepper was used as material, the effects of seed priming with various concentrations of GABA(0, 1.0, 2.0, 4.0, 6.0, 8.0 μmol·L^-1)on plant biomass, leaf osmotic regulating substance, leaf antioxidant capacity, leaf photosynthesis system, potassium and sodium ion uptake and distribution between leaves and shoots of pepper plants under 100 mmol·L^-1 NaCl stress applied at 4-6 leaf stage were investigated. The results were as follows:(1)From the point of plant growth under salt stress, the best concentration of GABA for seed priming of pepper was 6.0 μmol·L^-1, which greatly boosted the biomass of pepper plants under salt stress.(2)The mechanisms of GABA seed priming promoting salt tolerance of pepper were further analyzed. Seed priming increased the leaf contents of soluble sugar, soluble protein, chlorophyll and proline, decreased the leaf contents of O₂^- and malondialdehyde(MDA), enhanced the leaf activities of antioxidant enzymes, including super oxide dismutase(SOD), peroxidase(POD), catalase(CAT)and ascorbate peroxidase(APX), and raised several chlorophyll fluorescence metrics, including F_v'/F_m', qP_Lss, QY_Lss, NPQ_Lss and Rfd, reduced the K⁺ content and K⁺/Na⁺ ratio in roots and stems.(3)For a comprehensive understanding of the mechanisms of GABA seed priming promoting salt tolerance of pepper, grey correlation analysis was carried out. Based on results of grey correlation analysis, seed priming with GABA significantly alleviated salt stress to pepper plants by boosting the activities of the antioxidant enzymes POD and CAT as well as increasing the levels of osmotic regulators. In conclusion, seed priming with 6.0 μmol·L^-1 GABA significantly increases salt tolerance of pepper seedlings, probably by improving antioxidant and osmotic regulating capacities of pepper plants.

Effects of different application modes of phosphorus- solubilizing bacterial fertilizer on seedling root growth and phosphorus nutrition of Pinus massoniana families

2023/12/30 21:36:09
In order to study the effect of different application modes of phosphorus-solubilizing bacterial fertilizer(PSB fertilizer)on seedling phosphorus nutrition of Pinus massoniana, a pot inoculation experiment with local and evenly application of PSB fertilizer was set up. We studied the responses of P uptake and root growth of different P. massoniana families under different PSB fertilizer treatments through using WinRHIZO Pro STD1600+ root image analysis software and Molybdenum antimony colorimetric method. The results were as follows:(1)Inoculating PSB fertilizer had a significant effect on growth of P. massoniana seedlings. The main growth indexes, such as seedling height, ground diameter, root dry matter weight, root/shoot and whole plant dry matter weight, significantly increased under local treatment.(2)Local treatment significantly increased the root length, root surface area, root volume and root tip number of seedlings. The root length of 0 P. massoniana showed different performance under application of PSB fertilizer. The low phophorus tolerance provenance No. 22 was a much sensitive family, and all growth indexes were significantly higher than those of No. 10 and No. 50 families. In conclusion, local application of PSB fertilizer significantly promoted the growth and P uptake of P. massoniana families under low P environment, and this study provides guidances for the management of P. massoniana plantation under P poor sites.

Effects of light quality and photoperiod on growth and physiology of Michelia baillonii seedlings

2023/12/30 21:36:09
In order to explore an efficient artificial light environment for improving the quality of Michelia baillonii and shortening its cultivation cycle, the annual seedlings of M. baillonii were exposed to five composite lights, red+ blue(8R1B, 6R1B), red+blue+purple+green(8R1B1P1G, 6R1B1P1G), and white light(W)with two photoperiods(12,16 h·d^<sup>-1), two-factor experiment with randomized block design and subordinate function were used to explore the response pattern of the growth, photosynthetic pigments and endogenous hormone contents of M. baillonii seedlings to different light qualities and photoperiods. The results were as follows:(1)Light quality, photoperiod, and their interaction had significant effects on height growth, leaf area, chlorophyll a, zeatin(ZR), abscisic acid(ABA)content, and endogenous hormone ratio(IAA/ABA,(IAA+GA₃+ZR)/ABA)(P<0.05).(2)16 h·d^-1 photoperiod was conducive to the improvement of height growth, leaf area, seedling quality index, biomass, chlorophyll a, auxin(IAA), ZR content, and endogenous hormones ratio.(3)Under 16 h·d^-1, height growth, leaf area, and seedling quality index under 8R1B treatment reached the maximum value, which were 21.84 cm, 158.39 cm² and 2.43, respectively; compared with the 6R1B treatment, the 8R1B had higher chlorophyll a/b and ZR content; compared with the 6R1B1P1G treatment, the 8R1B1P1G had higher chlorophyll a, a/b, carotenoids, IAA, gibberellin(GA₃)and endogenous hormones ratio. In conclusion, red-blue composite light quality with higher red ratio and proper extension of photoperiod are conducive to improving the quality of M. baillonii, while the addition of purple-green light can not promote its growth, 16×8R1B is the most suitable light condition for the growth of M. baillonii seedlings.

Genetic diversity of six Amorphophallus species in Southwest China based on cpDNA sequences

2023/12/2 0:00:00
Investigating mechanisms underlying genetic variation and differentiation of wild resources is critical for the utilization and improvement of crop germplasms. Amorphophallus species is an important economic crop in Southwest China for its glucomannan production. However, the wild populations of this genus are declining due to human activities. To investigate the genetic diversity and phylogenetic relationship of representative Amorphophallus species in Southwest China, the phylogenetic relationships between species were reconducted by using three chloroplast DNA(cpDNA)fragments and analyzing genetic diversity of six Amorphophallus species. The results were as follows:(1)The genetic diversity of wild Amorphophallus populations was generally low with an average haplotype diversity(H_d)of 0.428. In addition, nearly half of the total populations had only one haplotype. The haplotype diversity of each species varied from 0.704 to 0.983.(2)The genetic differentiation between each pair of six species was relatively high, and the genetic differentiation coefficient(F_ST)values ranged from 0.481 to 0.967.(3)The phylogenetic analysis suggested that 27 selected Amorphophallus species should be mainly divided into three clades Africa clade, Southeast Asia clade, and East Asian continent clade. A. paeoniifolius belonged to the Southeast Asia clade. The East Asian continent A clade included A. konjac and A. krausei, and East Asian continent B clade was comprised of A. kiusianus, A. yunnanensis and A. tonkinensis. Geographic isolation and human disturbance could have caused the low genetic diversity in wild populations of Amorphophallus. The divergence of the East Asian continent clade may be driven by the rapid radiation and ecological adaptation in species of this clade. These findings provide theoretical guidance for the conservation, sustainable utilization and breeding of Amorphophallus species in Southwest China.

Transcriptome characteristic analysis and EST-SSR marker development of Capsicum annuum under waterlogging stress

2023/12/2 0:00:00
SSR molecular markers were obtained based on abundant transcriptome data under different waterlogging conditions in order to develop appropriate biological tools and explore the molecular mechanism of hot pepper response to waterlogging stress in this experiment. The results were as follows:(1)A total of 128 939 Unigenes were obtained from transcriptome analysis of hot pepper. The total length, average length, and GC content were 55 082 725 bp, 1 101 bp and 40.57%, respectively. Compared with seven major functional databases, 102 123(NR: 79.20%), 110 157(NT: 85.43%), 70 203(SwissProt: 54.45%), 73 539(KOG: 57.03%), 77 646(KEGG: 60.22%), 77 442(GO: 60.06%)and 68 216(Pfam: 52.91%)Unigenes obtained functional annotations. It was found that lipid metabolism, carbohydrate metabolism, amino acid metabolism, environmental adaptation, secondary metabolite biosynthesis, signal transduction and translation played important roles in the response to waterlogging stress.(2)A total of 26 574 SSR loci were found in 24 889 Unigenes from hot pepper transcriptome data. The frequency of SSR was 20.61%, of which single nucleotide repeat accounted for the highest ratio(37.26%), followed by tri-nucleotide(31.00%)and di-nucleotide(25.44%)repeat types, accounting for 93.70% of the total EST-SSR. The motifs A/T, AG/CT and TC/GA, followed by AT and TA, were the most abundant in mono-nucleotide and di-nucleotide. The most common types of tri-nucleotides were TTG/CAA and ACA/TGT.(3)10 002 pairs of EST-SSR primers were designed with Primer 3. A total of 30 pairs of primers were randomly selected for amplification test, and all of them were effective. Seven pairs of primers could amplify the target bands in three hot pepper germplasms. To sum up, the motif structure of the most dominant SSR repeat type in hot pepper is basically similar to that of other varieties, and the molecular mechanism of hot pepper waterlogging stress response is preliminarily explored. The EST-SSR markers are developed to provide reference for hot pepper waterlogging-tolerant genetics breeding.

Genetic diversity and structure of wild Rosa roxburghii in Southwest China

2023/12/2 12:35:43
In order to study genetic diversity and the origin of wild Rosa roxburghii in Southwest China and provide some bases for its utilization and protection. A survey was based on the splicing sequences of two single-copy nuclear genes(GAPDH and ncpGS)and three chloroplast genes(atpF-trnH, trnL-trnF and trnG-trnS). The individuals of 320 wild R. roxburghii from 27 wild populations in China were amplified, sequenced by PCR. After that, the sequencing results were analyzed with relevant software. The results were as follows:(1)Low genetic diversity levels were found in R. roxburghii of single-copy nuclear gene(Hd=0.469 2, π=0.000 49)and chloroplast gene(Hd=0.653 4, π=0.000 65). But there were significant differences among different populations.(2)Analysis of molecular variance(AMOVA)showed that the genetic variation almost occured within populations, which indicated that population variation was the main source of genetic variation in wild R. roxburghii. It existed obvious genetic differentiation among populations(cpDNA: F_ST=0.336 47, G_ST= 0.273, N_ST= 0.308; scnDNA: F_ST=0.094 87, N_ST=0.076, G_ST=0.056). The distribution of R. roxburghii did not have obvious phylogeographical structure(P>0.05).(3)Tajima's D value of neutral test was insignificantly negative value, indicating that R. roxburghii populations conformed to neutral evolution model. Fu's Fs value was significantly negative, combining the result of mismatch analysis curve, deducing that R. roxburghii populations had an expansion within a small range before. But generally, they remained stable.(4)According to the haplotype network, the populations of the Bijie region not only presented higher genetic diversity, but also had a lot of haplotypes. Therefore, the Bijie region was speculated to be one of the refuges of ice age. Therefore, they were supposed to be carried out the strategy of local protection. The populations with special traits and unique haplotypes shall be protected to adopt a priority protection approach. The survey is expected to provide a reference for wild R. roxburghii of the resource protection and genetic breeding.

Analysis of gene expression characteristics before and after seed germination of Polygonatum cyrtonema

2023/12/2 12:35:43
Polygonatum cyrtonema seeds have a comprehensive dormancy phenomenon, and analyzing the changes of key genes before and after seeds germination plays an important role in exploring seed physiology and breaking dormancy. Based on high-throughput sequencing technology(Illumina Hiseq 2500), the transcriptome sequencing and bioinformatics analysis were carried out in four stages of P. cyrtonema seed germination with three biological replicates. The results were as follows:(1)388 231 Transcripts and 178 319 Unigenes were obtained by de novo assembly.(2)Among the 11 817 Unigenes with significant differences before and after germination, 6 405 were up-regulated and 5 412 were down-regulated.(3)Significant difference genes were enriched and analyzed in the GO enrichment analysis, and the differential expression up- and down-regulation Uningenes were main enriched in biological process and molecular function, which were mainly involved in metabolic processes and catalytic activities.(4)KEGG significant enrichment indicated that the differential expression genes were mainly enriched in ribosome, plant hormone signal transduction, starch and sucrose metabolism and other pathways; the up-regulated differential genes were mainly enriched in ribosome 231, plant hormone signal transduction 56, starch and sucrose metabolism 76 and others; the down-regulated differential genes were mainly enriched in phenylpropanoid biosynthesis 48 plant hormone signal transduction 48, starch and sucrose metabolism 48, and other pathways; a total of 40 differential genes were involved in the synthesis of auxin pathway key enzymes. Genes encoding sucrose phosphate synthase were down-regulated and genes encoding glycogen phosphorylase were up-regulated in starch and sucrose metabolic pathways. This paper preliminarily clarified that plant hormone signal transduction, starch and sucrose metabolism played a key role before and after the seeds germination of P. cyrtonema and analyzed the key genes involved in these two pathways. This study preliminarily clarifies that plant hormone signal transduction and starch and sucrose metabolism play a key role before and after the germination of P. cyrtonema seeds, and analyzes the key genes involved in these two pathways, which provides a reference for further research on the physiology, reproduction and breeding of P. cyrtonema seeds.

Genetic diversity analysis and comprehensive evaluation of phenotypic characters in 20 germplasm resources of Erythropalum scandens

2023/12/2 12:35:43
In order to analyze the phenotypic character of different germplasm resources of woody vegetable Erythropalum scandens and lay a foundation for selecting excellent E. scandens resources with large leaf and vigorous branch. In this study, 20 provenances from Vietnam and three provinces(regions)in China(Guangxi, Guangdong and Fujian)were selected as the research objects. Twelve leaf character and four branch characters were measured and calculated. Descriptive statistics, variance analysis and correlation analysis were performed, and the phenotypic characteristics of different E. scandens resources were counted, classified and evaluated. The results were as follows:(1)There were significant or extremely significant differences in most leaf and branch characters among different resources. The rangeabilities of the coefficient variation(CV)of all characters within provenances were not the same. The rangeability order of the coefficient variation of all characters among different resources was leaf functional character(15.42%～70.01%)> branch character(20.57%-71.71%)> leaf morphological character(3.39%～20.01%). Phenotypic variation within provenance was more prominent.(2)In terms of correlation between leaf morphological character and functional characteristic, there was a significant correlation between number of new branches, number of internodes and number of new leaves, but there was no significant correlation between number of new branches and leaf morphological characters.(3)Four principal components could be extracted from 16 phenotypic characters, and the total contribution rate was 85.528%. Four principal components reflected leaf morphology, leaf germination and growth, leaf shape, dry matter accumulation and branch thickening, respectively.(4)Cluster analysis of 20 resources could be divided into three categories, one had large leaves and good growth condition, one had small leaves and vigorous branches, and the other one was not outstanding in comprehensive performance. The geographical distribution of the subgroups of the major resources was close to each other.(5)Anxi and Fuqing in Fujian could be selected as resources with large leaves; Daxin, Shangsi and Guiping in Guangxi could be selected as resources with strong branches. In conclusion, the provenances of Anxi in Fujian has the best comprehensive performance, followed by Fuqing, Haifeng, Nanning and Chaling. Zhaoping and Yizhou has the worst comprehensive performance and they are not suitable for cultivation in Nanning. In some resources, there are excellent single plants with outstanding growth performances, which could be developed into clones for further provenance tests. This study provides a scientific basis for the analysis of phenotypic characters and the initial performance of different E. scandens germplasm resources in Nanning, and lay a foundation for screening and breeding high-yield E. scandens varieties.

Phenotypic traits and SSR molecular identification of hybrid progenies of Camellia chekiangoleosa × C. semiserrata

2024/8/5 10:00:45
Camellia chekiangoleosa has high oil concentration and oleic acid content, while C. semiserrata has strong growth vigor and resistance. In order to make the utmost of the advantages of C. chekiangoleosa and C. semiserrata and cultivate excellent germplasm materials, the phenotypic traits of 45 F₁ hybrid progenies of C. chekiangoleosa and C. semiserrata were analyzed to grasp the phenotypic traits of the hybrid progenies. In addition, SSR markers were used to identify hybrids, and SSR markers that could be used to identify the hybrid progenies of oil tea were screened. The results were as follows:(1)The F₁ hybrids of C. chekiangoleosa × C. semiserrata showed tall tree and rapid growth, and their leaf veins, sepals and stigmas were all tended to the traits of the male parent C. semiserrata, while flower and leaf morphologies and other traits were similar to the female parent C. chekiangoleosa, and the characteristics of leaf color and size were between those of the parents.(2)From the 32 SSR markers, eight fully complementary markers that could distinguish parents and determine the origin of offspring were screened out for identification of hybrid progenies, among which the hybrid identification rates of seven markers were as high as 100%, and the hybrid identification rate of one marker was 55.56%. And 45 hybrid progenies were all true hybrids identified by the complementarity of eight markers.(3)The eight SSR markers were used to verify the ability to identify the hybrid progenies, indicating that it is feasible to use these SSR markers to identify the authenticity of the hybrid progenies of oil tea. This study results provide a reference for interspecies cross breeding of oil tea, and also provide a reference for the SSR marker identification of hybrids of oil tea.

Construction of a mutant library associated with aroma genes in tobacco using CRISPR/Cas9

2024/8/5 10:00:45
To explore the feasibility of constructing a tobacco mutant library using the CRISPR/Cas9 gene editing technology, we designed sgRNAs of 100 aroma related genes in tobacco, constructed a plasmid library composed of 100 corresponding CRISPR/Cas9 editing vectors, and analyzed the co-transformation rate, on-target editing rate, and off-target editing of transgenic offspring using the tobacco variety ‘Honghua Dajinyuan' as experimental material in this study. The results were as follows:(1)After co-transformation of 100 sgRNAs mediated by Agrobacterium tumefaciens, 77 of them were detected in 172 positive transformation strains, with a co-transformation rate of 77%.(2)Among 77 transgenic offspring carrying sgRNA, 69 sgRNAs edited the target genes, with an editing rate of 89.6%.(3)Sequencing detection revealed that only one sgRNA produced off-target editing at a non-target site, indicating a very low probability of off-target editing of CRISPR/Cas9 in tobacco. In summary, it is feasible to construct a mutant library by the co-transformation of a CRISPR/Cas9 vector library to edit a large number of candidate target genes in tobacco. This method has the characteristics of high co-transformation rate, high editing rate, and low probability of off-target editing.

Bioinformatics and expression analysis of expansin genes GmEXPB5 and GmEXPB7 in soybean

2024/8/5 10:00:45
Expansin(EXP)plays an important role in plant response to environmental stress by regulating cell wall relaxation. To explore the role of EXP genes in soybean response to abiotic stress, two soybean EXP genes(GmEXPB5 and GmEXPB7)and their protein sequences were analyzed by bioinformatics, and their expression levels were detected by real-time fluorescent quantitative PCR(qRT-PCR). The results were as follows:(1)The GmEXPB5 and GmEXPB7 were located on chromosomes 10 and 12 of soybean, and encoded proteins containing 272 and 267 amino acids, respectively. The molecular weight of GmEXPB5 protein was 29.07 kD and the theoretical isoelectric point was 7.51. The molecular weight of GmEXPB7 protein was 29.09 kD and the theoretical isoelectric point was 8.66. Both GmEXPB5 and GmEXPB7 were stable hydrophilic proteins, and localized in the cell wall. Both GmEXPB5 and GmEXPB7 proteins contained a signal peptide sequence and a conserved DPBB_1 structural domain.(2)GmEXPB5 protein had the closest affinity with CaEXPB15 protein of Cicer arietinum, and GmEXPB7 protein was closely related to EXPB3 proteins of Spatholobus suberectus, Vigna angularis and V. unguiculata.(3)GmEXPB5 and GmEXPB7 were expressed in root, stem and leaf of soybean, and their expression levels in root and leaf were significantly higher than those in stem.(4)GmEXPB5 and GmEXPB7 could respond to salt, drought and cold stresses in soybean seedlings.(5)The promoter region of GmEXPB5 contained two types of stress-related cis-elements(ABRE and ARE). The promoter region of GmEXPB7 contained five types of stress-related cis-elements(ABRE, ARE, CGTCA-motif, TC-rich repeats and MBS). In conclusion, these results indicate that GmEXPB5 and GmEXPB7 can participate in the response of soybean to abiotic stress.

Maintenance and variation of chromosome ploidy under tissue culture conditions of different Camellia germplasms

2024/8/5 10:00:45
The maintenance and variation of ploidy in plants are influenced by environmental factor, and the tissue culture regeneration can often cause changes of chromosome structures and ploidy levels. In order to explore the ploidy variation of Camellia germplasm under tissue culture conditions, the callus induction system of Camellia germplasm was used to analyze the ploidy changes by flow cytometry, and the stability and variation of the ploidy under tissue culture generation were analyzed in combination with colchicine treatment. The results were as follows:(1)Six of ten of Camellia germplasms were diploid, two of them were tetraploid, one hexaploid and one decaploid, and the germplasm materials largely maintained stable ploidy levels during the tissue regeneration processes.(2)The optimum conditions for colchicine treatment were obtained, that is, the callus was cultured in treatment medium containing colchicine at 20 mg·L^-1 for 10 d, followed by regeneration in tissue-differentiation culture.(3)The ploidy analyses were carried out on the treated tissues in different differentiation states, and the results showed that most of the independent tissues(including 56 callus and shoots)were found to be aneuploids, 38 tissues had ploidy between 1.5 to 2.5, and 11 tissues produced less than 1.5. This study further explores the ploidy relationship between different Camellia germplasms, and provides a reference for an in-depth study of ploidy regulation and polyploid induction of Camellia.

Influences of polyamines on embryogenic callus proliferation and somatic embryogenesis in Litchi chinensis

2024/8/5 0:00:00
To explore the effect of exogenous polyamines(PAs)on embryogenic callus(EC)proliferation and somatic embryogenesis of litchi, the morphology, structure, endogenous PA content and related enzyme activities of EC were systematically investigated using the ‘Feizixiao' ECs as materials subcultured on the medium supplemented with exogenous putrescine(Put), spermidine(Spd), and spermine(Spm). The results were as follows:(1)Exogenous Put, Spd and Spm treatments significantly increased the EC proliferation rate and reduced the amount of induced somatic embryos and number of germinations. The proliferated EC cells after exogenous PA treatments were more consistent in size and stained deeply and evenly. Furthermore, multicellular proembryos in EC were reduced, and fully differentiated early cotyledon embryos could be seen.(2)All the exogenous PA treatments significantly increased the endogenous PA content in EC. Among them, Put treatment had the highest content of each endogenous PA component and total PA. When the EC proliferated on the medium containing exogenous PAs was transferred to the medium without exogenous PAs(recovery culture)for proliferating, the Put content in the EC was still significantly higher than the control, however, the endogenous Spd and Spm were significantly decreased.(3)Exogenous Put treatment significantly increased the activities of ornithine decarboxylase(ODC), arginine decarboxylase(ADC), and diamine oxidase(DAO)in EC, while exogenous Spd and Spm treatments significantly reduced the activities of ODC and ADC in EC, and exogenous Spd significantly increased the polyamine oxidase(PAO)activity. When transferred to the recovery culture medium, the ADC and DAO activities of newly proliferated EC were significantly lower than those of EC cultured with exogenous PAs, but there was no significant difference in ODC and PAO activities. In summary, the exogenous PAs can affect endogenous PAs content by regulating the activities of enzymes related to PAs metabolism, thereby affecting EC proliferation and somatic embryo induction in litchi. These results provide a basis for further study on the mechanism of PAs regulating embryogenesis, and improve litchi regeneration efficiency in vitro.

Analysis of expression regulation of AP2 gene by pre-miR172 and miR172 in rape

2024/6/5 11:30:13
The appropriate flowering time is of great significance for crop yield. In the cropping pattern of “rice-rice-oil” in southern region, researching on the flowering period of Brassica napus can provide a theoretical basis for the breeding of early maturing rape varieties. The AP2 family transcription factors in rape are widely involved in the growth and development and play an important role during flower development. However, there are few studies exploring the regulation of AP2 at the microRNA level. In order to investigate the regulatory functions of the miR172 precursor(pre-miR72)and mature body(miR172)on AP2 gene in rape, the regulatory elements of miR172 and AP2 promoters were predicted based on bioinformatics, then the evolutionary relationship of six rape AP2 genes and the targeting relationship between miR172 and AP2 were analyzed, and the expression patterns of AP2, miR172 and pre-miR172 in different tissues of early and late maturing rape were detected by qRT-PCR. Finally, the correlation between miR172 abundance and AP2 expression level was studied, as well as the correlation between pre-miR172 and miR172. The results were as follows:(1)Both miR172 and AP2 promoter regions had cis-elements that regulated flower development.(2)The six AP2 sequences holded the strong purification selection, and they were the target genes of miR172 based on their binding sites for miR172.(3)miR172 family could promote the flowering of early maturing rape by increasing AP2 expression levels, except for miR172d. In late maturing rape, miR172a and miR172c performed weakly in flowering, while miR172b and miR172d worked together to reduce the expression level of AP2 to inhibit flowering.(4)The pre-miR172 family had a promoting effect on the expression level of miR172 family in early maturing rape; in late maturing rape, pre-miR172a and pre-miR172b exerted positive regulation on the formation of their mature bodies, while pre-miR172c and pre-miR172d exerted the opposite effects. After overexpression of pre-miR172, the expression patterns of miR172 and AP2 remained consistent with the above results, confirming the regulatory function of pre-miR172 on miR172 and AP2. The results of this study enrich the functional regulation pathway of rape AP2 gene, and provide new ideas for the study of gene regulatory function.

Molecular cloning, location and promoter analysis of NDH dehydrogenase gene from Santalum album

2024/6/5 11:30:13
In order to investigate the function and regulation mechanism of NDH dehydrogenase gene in Santalum album, the technique of RACE was used to amplify the full-length sequence of SaNDH6 with heartwood as material. The technique of quantitative real-time fluorescence PCR(RT-qPCR)was employed to analyze its expression in different tissues and after hormone induction. The subcellular location was determined by Arabidopsis thaliana protoplast transient expression. 2 kb cis-acting element upstream of start codon ATG was analyzed by PlantCARE online service, and the transcription factors which could bind the cis-acting elements was predicted by PlantRegMap software. The results were as follows:(1)SaNDH6 encoded 303 amino acids. It was a hydrophobin and located in chloroplast.(2)The phylogenetic tree analysis indicated that SaNDH6 had a more closely evolutionary relationship with NDH6 from woody plants.(3)Plant care analysis showed that the promoter sequence of SaNDH6 contained a large number of light responsive cis-acting elements such as ACE, AE-box, Box 4, G-Box and GT1-motif. It also contained abscisic acid(ABA)responsive element ABRE, jasmonic acid methyl ester(MeJA)responsive elements CGTCA-motif and TGACG-motif, gibberellin(GA₃)responsive elements P-box, ARE cis-acting regulatory element essential for the anaerobic induction, and TC-rich repeats element involved in defense and stress responsiveness.(4)The results of plantRegMap analysis showed that there were 76 transcription factors that could bind to the SaNDH6 promoter, and among which, ERF transcription factor was the most(up to 40 TFs).(5)SaNDH6 can be expressed in the tissues of roots, heartwoods, calluses and leaves, but had a higher expression level in the tissue of leaves; under 1×10^-4 mol·L^-1 MeJA and GA₃ treatments, the expression level of SaNDH6 were significantly elevated after 3 h when compared with 0 h, respectively. In conclusion, SaNDH6 was a nucleus gene encoding protein, its expression was induced by light and some hormones, and it might be involved in against some defense and stress processes in S. album.

Evaluation and analysis of pine resin components of Pinus elliottii clones

2024/3/3 12:56:19
In order to select superior clones with strong resin-producing capacity and high resin-quality from the existing fast-growing timber clones in seed orchard of Pinus elliottii, and to make high use of crop germplasm resource, 36 clones in the first generation of slash pine seed orchard were used as materials to determine their resin yield, resin mass flow rate and DBH growth, and to analyze their resin composition by GC-MS. Based on the above indicators, correlation analysis and cluster analysis was used to comprehensively evaluate the production and quality of resin among 36 clones. The results were as follows:(1)There were a total of 21 pine resin components, including 8 monoterpenes and 13 diterpenes.(2)Correlation analysis showed that the resin mass flow rate(RMR)had significant and positive correlation with the total content of monoterpene, weakly negatively correlates to abietic-type resin acid, and not significantly correlated to pimaric-type resin acid.(3)Based on the cluster analysis results integrating four types of indicators as the total monoterpene content, resin mass flow rate, abietic-type resin acid and pimaric-type resin acid, 36 clones could be divided into three categories and the difference between each type was significant. The performance of Class 1 was much better than that of the other two categories.(4)There were 17 high-resin yield pine clones(ERM≥15.15)among 36 clones, and on the basis of this, four clones(6-44, 4-11-1, 1-38, 3-64)display higher monoterpenes content, while four clones(4-11-1, 3-64, 2-0420, 3-468)showed higher contents of pimaric-type resin acid. And the content of abietic-type resin acid of clone 2-173 was the highest. In summary, a total of 21 pine resin components of P. elliottii were identified, and 36 clones were evaluated based on four indicators: the total monoterpene content, resin mass flow rate, abietic-type resin acid and pimaric-type resin acid. We not only analyzed qualitatively the resin composition but also evaluated quantitatively the resin-producing capacity of 36 clones in slash pine seed orchard. Our findings provide the scientific references for the targeted breeding of pine resin components and lay a foundation for subsequent heredity breeding and gene improvement of P. elliottii.

Cloning of HDR gene in Pinus massoniana and Cloningits response to drought and salt stresses

2024/3/3 12:56:19
Drought and land salinization are inhibiting factors for the sustainable development of forestry, and the plants will release volatile substances such as terpenoids during biological or non-biological stress. The 1-hydroxy-2-methyl-2-(E)-buteny-4-pyrophosphate reductase is the terminal active enzyme of the MEP pathway, which provides precursor terpenoids and has the main rate-limiting effect. In order to investigate whether HDR gene of Pinus massoniana is involved in stress response under drought and salt stress, the open reading frame of PmHDR gene was cloned, and bioinformation, tissue specific expression and preliminary function were analyzed. The results were as follows:(1)The coding region length of PmHDR gene is 1 458 bp, encoding 485 amino acids, and its encoded protein contains the core sequence of LytB/IspH gene superfamily and PLN02821 multifunctional structural domain, which belongs to the HDR family.(2)The PmHDR codon use preference was weak, with a preference for codons ending in A/U. Nicotiana tabacum, Arabidopsis thaliana and Saccharomyces cerevisiae were more suitable as its heterologous expression receptors.(3)Results of qRT-PCR showed that the PmHDR gene was most highly expressed in old needles of Pinus massoniana, followed by young needles, young stem and old stem and least expressed in root.(4)The gene expression vector pBI121-PmHDR was constructed and transformed into Arabidopsis thaliana. The transgenic A. thaliana showed greater resistance to drought and salt stress. These results indicate that PmHDR is involved in plant response and regulation to drought and salt stress, and provide some theoretical supports for Pinus massoniana stress-resistance breeding.

Correlation of HrANR genes and flavonoid accumulation with drought resistance in sea buckthorn(Hippophae rhamnoides subsp. sinensis)

2024/3/3 12:56:19
Anthocyanidin reductase is one of the key enzyme involved in the synthesis of flavonoids. In order to explore the structure of ANR gene, ANR enzyme expression pattern and flavonoid content under drought stress and their correlation, a HrANR gene identified from RNA-seq data of sea buckthorn was screened and analyzed by bioinformatics soft, the expression patterns of HrANR gene in different tissues and of flavonoid contents in leaves were performed. The results were as follows:(1)The ORF of HrANR gene was 1 017 bp, which encoded 338 amino acids. It was a stable hydrophilic protein, and the homologous protein had significant family and genus characteristics.(2)HrANR gene was expressed in roots, stems and leaves of sea buckthorn under drought stress, but the expression trends were different, with an increase followed by a decrease and then an increase in roots, a continuous decrease in stems, and an increase followed by a continuous decrease in leaves.(3)The flavonoid contents in leaves of sea buckthorn under different levels of stress showed a trend that first increased continuously and then decreased slightly, increased to the highest point after rehydration. The above results indicated that the expression of HrANR gene and the changes of flavonoid content were closely related to the drought resistance of sea buckthorn. The flavonoid content in leaves was positively correlated with drought stress at the beginning of drought stress and negatively correlated with drought stress under severe stress.(4)Leaf expression, stem expression and flavonoid content of HrANR gene were negatively correlated(P_leaf= -0.751, P_stem= -0.934)and root expression was positively correlated with flavonoid content(P_root= 0.444). The above results indicate that the expression of HrANR genes and changes of flavonoid content in sea buckthorn are closely related to drought resistance, and it provides a reference for elucidating the drought resistance mechanism of sea buckthorn.

Construction of yeast two-hybrid cDNA library in cambium tissue of Hevea brasiliensis and screening of HbHDA6 interacting proteins

2024/3/3 12:56:19
The secondary laticifer is the position for synthesis and storage of natural rubber(NR), which is differentiated from the vascular cambium cells of bark in stem of rubber trees(Hevea brasiliensis). The quantity of secondary laticifer is depended on the frequency of the secondary laticifer differentiation from cambia, which is the main index of yield breeding of rubber tree. In previous studies, we found trichostatin A(TSA), an inhibitor of histone deacetylase(HDA), can also induce laticifer differentiation, and the histone deacetylase gene(HbHDA6)is a participator in laticifer differentiation. Because of the molecular mechanism of secondary laticifer differentiation regulated by histone acetylation has not been clarified. Therefore, we construct a yeast two-hybrid cDNA library used the vascular cambium tissues treatment by coronatine(COR), and screening the yeast two-hybrid library by HbHDA6 gene as the bait, for determining the proteins interacting with HbHDA6. The results were as follows:(1)The homogenized yeast two-hybrid cDNA library of vascular cambium was constructed by the technology of Gateway. The capacity of the primary library was 6.34 × 10⁶ CFU·mL^-1, the total number of clones was 1.27 × 10⁷, and the capacity of secondary library was 7.72 × 10⁶ CFU·mL^-1, the total number of clones was 1.54 × 10⁷, and the recombination rates of two libraries were 100%. The average length of inserted fragments was 1.1 kb and 1.2 kb in primary and secondary library, respectively.(2)The bait vector of pGBKT7-HbHDA6 for screening the proteins interacting with HbHDA6 was successfullyconstructed and confirmed no self-activation activity.(3)The pGBKT7-HbHDA6 bait vector was used to screen the constructed yeast two-hybrid cDNA library, and 22 proteins interacting with HbHDA6 were obtained by NCBI_BLAST comparison and removing duplicates, including CLP1, ERF3, ERF4, HSP82, LARP6a, APT5, PP2A, APT5, FBA6, etc. The results provide a theoretical basis for analyzing the molecular regulatory network of the secondary laticifer differentiation of rubber tree, and provide candidate genes for the rubber production potential of genetically modified and a new clue for the genetic improvement and breeding of high-performance NR.

AP3 gene cloning and gene-editing vector construction of Hydrangea macrophylla ‘Dooley'

2024/3/3 12:56:19
Hydrangea macrophylla is a garden plant widely cultivated in Asia, America, and Europe with its inflorescence as main ornamental feature. It is commonly used in interior decoration and landscape creation. To investigate the role of AP3 gene in hydrangea during calyx formation, H. macrophylla ‘Dooley' was used as the material. The MADS-box class B gene HmAP3 was cloned, and its gene function was predicted by bioinformatics analysis. To explore methods for quicker breeding new varieties, highly-specific editing targets were screened and CRISPR/Cas9 gene-editing vectors were constructed. The vector sequence was integrated into the H. macrophylla genome by agrobacterium-mediated transformation. The results were as follows:(1)The cDNA sequence full length of HmAP3 was 546 bp, encoding 181 amino acids. Its amino acid sequence was 100% similar to the reference sequence and 58.8% similar to Arabidopsis thaliana.(2)AP3 differed greatly in different genera. Within the same genus, the main structure of AP3 protein was conserved and differed only in a few motifs.(3)There were two highly specific targets in HmAP3. Sequencing results indicated that two single-target CRISPR/Cas9 gene-editing vectors were constructed successfully.(4)There were five resistant buds with Cas9 sequences in their genomes. However, their target sequences did not change due to the absence of Cas9 expression. In this study, the potential of AP3 gene in the breeding work of double flower phenotype was investigated, and a preliminary exploration of CRISPR/Cas9 gene-editing technology for Hydrangea macrophylla was conducted. These results provide a basis for the breeding of H. macrophylla.

Cloning and expression analysis of BjGSTF12 in Brassica juncea

2024/3/3 0:00:00
To explore the role of glutathione S-transferase gene(GST)in the accumulation of anthocyanin in Brassica juncea, one GST gene related to the anthocyanidin accumulation was cloned from near-isogenic lines of B. juncea with purple stalk and green stalk, and named as BjGSTF12. In this study, the bioinformatics characteristics of BjGSTF12 encoding protein and promoter were analyzed, the expression level of BjGSTF12 and the relationship with total anthocyanidin content were analyzed in B. juncea lines with purple stalk and green stalk. The results were as follow:(1)BjGSTF12 genes from B. juncea were successfully cloned, whose the full length of BjGSTF12 in genome and cDNA was 808 bp and 651 bp, encoding a protein of 216 amino acids. The BjGSTF12 contained the typical domains of GST proteins, namely GST_N and GST_C. However, their sequences of BjGSTF12 did not exhibit any differences between the two lines of B. juncea.(2)BjGSTF12 was closely related to AtGSTF12 in Arabidopsis, and both belonged to the φ subfamily of GST family.(3)The BjGSTF12 promoter sequences were cloned from two Brassica juncea strains of green stalk and purple stalk, and they exhibited four base mutations/insertions. However, the types and numbers of cis-acting elements did not show obvious differences between the two strains, including nine MYB transcription factor binding sites, one hormone response element, and three abiotic corresponding elements.(4)The total anthocyanidin content in B. juncea of purple stalk was significantly higher than that in green stalk ones, and the expression levels of BjGSTF12 in two lines were found to be similar to the total anthocyanidin content in both lines.(5)Protein interaction network analysis revealed that BjGSTF12 may interact with the key enzymes of anthocyanidin biosynthesis, glycosylation modification, and transporter proteins. In summary, BjGSTF12 is likely to play a key role in the accumulation of anthocyanidin in B. juncea by regulating its biosynthesis, modification, and transportation through interactions with other proteins. This study provides a theoretical reference for further study on the function of GST and the mechanism of anthocyanidin accumulation in B. juncea.

Metabolomics analysis of flower color substances in three Rosa rugosa cultivars

2024/3/3 0:00:00
Rosa rugosa is a deciduous shrub belonging to Rosa L. in Rosaceae. It has a high ornamental value and commercial value, but its single color limits the development and utilization of rose and its application in landscape architecture. In order to explore the coloring substances of three different varieties of roses, ‘Rosa rugosa &#215; Rosa sertata', ‘Rosa Crimson Glory' and ‘Rosa alba', this study used ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)to detect the types and contents of flavonoids in petals. The KEGG database was used to enrich the differential metabolites, screen out the key metabolites, and analyze the correlation with the phenotypic value of flower color. The results were as follows:(1)A total of 58 metabolites were detected in the petals of three different color rose varieties, of which only one anthocyanin was cyanidin-3-O-glucoside, accounting for 30.45%.(2)K-means clustering analysis showed that a total of 12 key metabolites were annotated to the KEGG metabolic pathway. Among them, pinocembrin and myricetin were the main substances that determined the red color of ‘Rosa rugosa &#215; Rosa sertata' and ‘Rosa Crimson Glory', and eriodictyol, luteolin and kaempferol were the main substances that determined the white color of ‘Rosa alba'. In conclusion, this study can provide a theoretical basis for the breeding of roses with specific colors and promote the application of roses in landscaping.

Genetic variation analyses of quality and agronomic traits of 26 Fagopyrum tatari-cymosum lines

2024/3/3 12:56:19
Fagopyrum tatari-cymosum is a semi-perennial new buckwheat type developped from the hybridization between F. tataricum and F. cymosum. To explore the genetic laws of agronomic and quality traits of F. tatari-cymosum, 26 lines of F. tatari-cymosum were selected as materials, and their quality traits and agronomic traits were analyzed by variance analysis, correlation analysis and cluster analysis. The results were as follows:(1)The variation coefficient for quality traits of F. tatari-cymosum was gliadin content > glutenin content > flavonoids content > total protein content > globulin content > albumin content>starch content.(2)The variation coefficient for agronomic traits of F. tatari-cymosum was branch number of main stems > node number within 20 cm of the bases > main stem diameter > grain area > 1 000-grain weight > node number of main stems > plant height > grain length to width ratio > grain width > grain length > shell rate > grain perimeter > grain diameter.(3)In the correlation analysis, the flavonoid content was significantly positively correlated with albumin content; the gliadin content was significantly or extremely significantly positively correlated with the main stems diameter, the node number of main stems, the node number within 20 cm of the bases, and significantly or extremely significantly negatively correlated with the 1 000-grain weight, grain area, grain perimeter, grain width, and grain diameter; the starch content was positively correlated with grain area, grain length and grain diameter.(4)By cluster analysis, 26 F. tatari-cymosum lines were divided into three groups. Group I belonged to high starch, short stem, multi branched, low shell rate, large long grain lines, which could be used as parent material for breeding purposes of high starch and low shell rate; Group Ⅱ belonged to high protein, high stem, thick, small grain type lines, which could be used as a material for breeding purposes of high protein and strong stress resistance; Group Ⅲ belonged to high quality, high yield, large grain type lines. The results provide theoretical references for the breeding of F. tatari-cymosum.

Identification and expression analyses of O-acyltransferase WSD genes in Rehmannia glutinosa

2024/3/3 12:56:19
Plant wax ester synthase catalyzes the synthesis of wax esters from long-chain alcohols and fatty acids, and plays very important roles in plant wax synthesis and some resistances to drought, pathogenic bacteria, ultraviolet radiation, cold and insect invasion and other environmental stresses; cadmium(Cd)is one of the toxic heavy metals with the highest content in environment, and seriously threatens plant growth, development, quality, yield, and plant food safety. In order to explore the Cd stress expressions of wax ester synthase genes in Rehmannia glutinosa, we identified its wax ester synthase genes from its full-length transcriptome sequencing data, analyzed both physiochemical properties, phytogenetic trees and conserved domains with bioinformatics methods, and tissue expressions and Cd stress expressions using qRT-PCR. The results were as follows:(1)Two wax ester synthase genes, named as RgOATWSD1 and RgOATWSD2, were identified, whose coding proteins were unstable hydrophobic proteins with amino acid lengths of 463 aa and 473 aa, isoelectric points of 8.86 and 9.34 and molecular weights of 51.31 kD and 52.49 kD, respectively.(2)Both proteins contained a conserved acyl_WS_DGAT domain and DUF1298 superfamily, in which the former accounted for 92.65% to 94.50% of the amino acid sequence.(3)Both proteins were located in the endoplasmic reticulum and both secondary structures were mainly composed of random coil and α-helix; RgOATWSD2 was not transmembrane protein but RgOATWSD1.(4)Both were differentially expressed in the roots, stems and leaves of R. glutinosa plants.(5)Both expressions were highly responsive to Cd stress, but both expression change trends were different under Cd stress. This study identifies two wax ester synthase genes in response to Cd stress, and lays a foundation for further research on Cd stress expression and other functions of RgOATWSD.

Bacterial wilt resistance gene Slmlo1/6 mutants in tomato created by CRISPR/Cas9 technology

2025/1/7 10:23:14
Bacterial wilt is a devastating soil-borne disease in tomato(Solanum lycopersicum)production. The pathogenic species are complex and tend to have a variation, while mlo caused by the recessive mutation of MLO genes has a broad-spectrum resistance. The previous study suggested that Slmlo1/6 may be involved in the resistance response to bacterial wilt in tomato. In order to further study the gene function of Slmlo1/6 in tomato bacterial wilt resistance, the genetic mutant plants were created by CRISPR/Cas9 technology and their phenotypes were identified followed. The results were as follows:(1)gRNA sequences of SlMLO1/6 were designed and assembled with the U6 promoters, then U6-gRNA1/6 fragments containing highly effective targets were ligated to CRISPR vector of pBGK via restriction enzyme Bsa I digestion, to construct the two-gene fusion knockout vector of pBGK-SlMLO1/6. The recombinant plasmid of pBGK-SlMLO1/6 was transformed into Escherichia coli DH5α competent cells and positive monoclonal clones were selected via plate cultivation. Using Agrobacterium tumefaciens GV3101 strains-mediated genetic transformation and resistance screening to hygromycin, a total of nine edited tomato plants were obtained with sequencing validation.(2)Target region sequencing showed that M2 and M8 plants had the 177 bp and 7 bp deletion of SlMLO1, respectively, M7 had the 12 bp deletion of SlMLO6, and M9 had a single base T insertion of SlMLO6. Except for four single gene homozygous mutants above, the other mutations were heterozygous.(3)RT-qPCR showed that compared with the wild type plant, SlMLO1/6 gene expression of the mutants was significantly decreased, especially M2, M7, and M8 plants.(4)Phenotypic identification indicated that SlMLO1/6 might be tomato bacterial wilt susceptibility genes. In conclusion, the knockout vector is successfully constructed for resistance MLO genes and tomato transformation is also achieved, homozygous mutants acquire resistance to bacterial wilt. Amino acid deletion and frameshift mutation may be the crucial reasons for the gene function change of Slmlo1/6 in resistance. The results provide a theoretical reference and genetic engineering materials for the gene function study in resistance to bacterial wilt and disease resistance breeding application of tomato.

Identification and Fusarium wilt resistance evaluation of distant hybrid offspring of Lilium brownii var. viridulum

2025/1/7 10:23:14
During the cultivation process of Lilium brownii var. viridulum, it is easy to be infected with wilt disease, while some varieties of ornamental lilies have strong resistance to wilt disease. The study aimed to create hybrids with improved resistance to wilt disease by crossbreeding Lilium brownii var. viridulum with various ornamental lily varieties. Disease-resistant ornamental lily cultivars were specifically chosen for reciprocal crosses with Lilium brownii var. viridulum. The authenticity of the hybrids was confirmed through SSR molecular markers, and the resistance of the hybrid offspring to wilt disease was subsequently assessed. The results were as follows:(1)Lilium Asiatic hybrids such as ‘Black stone' had strong resistance to Fusarium wilt, while Lilium brownii var. viridulum and ‘Pink planet' had the weakest resistance to Fusarium wilt.(2)A total of 28 capsules were obtained from the orthogonal combination using Lilium brownii var. viridulum as the maternal parent, and 3 Lilium brownii var. viridulum × ‘Black stone' hybrid seedlings were finally obtained. The cross combination of Lilium brownii var. viridulum as the male parent obtained a total of 74 capsules, and ultimately obtained a hybrid seedling of ‘Strawberry event' × Lilium brownii var. viridulum.(3)SSR molecular marker identification results showed that 3 hybrid seedlings of Lilium brownii var. viridulum × ‘Black stone' were real hybrids, while 1 hybrid seedlings of ‘Strawberry event' × Lilium brownii var. viridulum was a false hybrid.(4)The 3 Lilium brownii var. viridulum × ‘Black stone' hybrid seedlings obtained had moderate resistance to Fusarium wilt. This study provides a reference for the distant hybridization of Lilium brownii var. viridulum and ornamental lilies, and proves that Lilium brownii var. viridulum hybrids with Fusarium wilt resistance can be obtained through hybridization. This study provides technical guidance for the distant hybridization of Lilium brownii var. viridulum with ornamental lily varieties, laying the groundwork for the breeding of new lily varieties with enhanced resistance to wilt disease in the future.

Cloning and expression analysis of OCPI2 gene in rice

2025/1/7 10:23:14
To investigate the function of protease inhibitor genes in rice defense against herbivorous insects, the coding sequence of the protease gene OCPI2 from the rice variety ‘Zhonghua 11' was cloned. The sequence characteristics of OCPI2 gene were analyzed, and a phylogenetic tree was constructed using bioinformatics software. Real-time quantitative PCR was utilized to examine the expression characteristics of OCPI2 gene under herbivorous insect feeding and plant hormone treatment. The results were as follows:(1)The coding region of OCPI2 gene was 219 bp, encoding a protein of 72 amino acids. The OCPI2 protein had a molecular weight of 7.72 kDa and a theoretical isoelectric point of 5.21, with no signal peptide and no transmembrane structure.(2)The OCPI2 protein was closely related to the homologous protein in Triticum urartu(EMS61613.1).(3)The OCPI2 gene contained a potato_inhibit conserved domain and belonged to the serine protease inhibitor family.(4)The expression of OCPI2 gene was induced by feeding from the rice striped stem borer(Chilo suppressalis)and the brown planthopper(Nilaparvata lugens), mechanical damage, and methyl salicylate treatment, whereas methyl jasmonate treatment consistently suppressed OCPI2 gene expression. Taken together, these results suggest that OCPI2 gene may be involved in the induced defense response of rice to herbivorous insects. This study provides a theoretical basis for a deeper understanding of the function of OCPI2 gene in rice defense against insect herbivores.

Physiological and biochemical responses of grape dormancy breaking with cyanamide and expression analysis of related genes

2025/1/7 0:00:00
To explore the responses of physiological and biochemical and mechanism of molecular regulatory for grape dormancy breaking with cyanamide, Vitis vinifera x V. labrusca ‘Shuijing' was used as experimental material in this study to determine the changes in activities of SOD, POD, CAT, contents of MDA and H₂O₂, and oxygen free radical production rate, and RT-PCR technology was used to clone the full-length cDNA sequences of two FT(Flowering location T)genes(VvFT1 and VvFT2)and one CBF(C-repeat Binding Factor)gene(VvCBF)from its buds, then their physicochemical properties, phylogenetic evolution, conserved motifs and domains, and expression levels differences in grape buds after cyanamide treatment were analyzed. The results were as follows:(1)The analysis of physiological and biochemical indicators showed that activities of SOD, POD and CAT, contents of MDA and H₂O₂, and oxygen free radical production rate in grape buds were significantly increased after treated with cyanamide.(2)The full-length cDNA sequences of VvFT1 and VvFT2 genes were 525 bp from Vitis vinifera &#215; V. labrusca ‘Shuijing', encoding 174 aa, and the full-length cDNA sequences of VvCBF gene was 826 bp, encoding 237 aa.(3)The homology analysis showed that VvFT1 of Vitis vinifera &#215; V. labrusca ‘Shuijing' had the highest amino acid homology with Litchi chinensis(LcFT: AEU08960.1)and Dimocarpus longan(DlFT2: ALA55998.1), VvFT2 had the highest amino acid homology with LcFT(AEU08961.1)and DlFT2(AHF27444.1). The phylogenetic analysis showed that VvFT1, VvFT2, LcFT(AEU08960.1, AEU08961.1)and DlFT2(ALA55998.1, AHF27444.1)clustered into a branch, with the most closes genetic relationship; VvCBF had the highest amino acid homology with Prunus ledebouriana(PlCBF: AEB69782.1), and the phylogenetic analysis showed that VvCBF and PlCBF clustered into a branch, with the most closest genetic relationship.(4)qRT-PCR analysis showed that VvFT1 and VvFT2 expression levels were significantly increased in buds after treated with cyanamide, while VvCBF expression level was significantly decreased. In summary, this study comprehensively analyzed the phylogenetic evolution of VvFT1, VvFT2, and VvCBF genes, as well as the expression patterns of these genes and physiological and biochemical indicators in grape buds after treated with cyanamide, laying a theoretical foundation for the molecular and physiological mechanisms of grape dormancy breaking with cyanamide.

Cloning and expression analysis of sugar transporter protein gene MeSWEET17b in cassava

2025/1/7 10:23:14
Cassava(Manihot esculenta)is an important food crop in tropical and subtropical regions. Sugar transporter protein SWEETs facilitate the flow of sugar between cells and play an important role in plant growth and development. In order to clarify the function of SWEET family genes in cassava, cassava‘KU50' was used as material in this study, and the gene properties of MeSWEET17b were studied by gene cloning, bioinformatics analysis, subcellular localization, in vitro yeast detection and RT-qPCR, etc. The results were as follows:(1)The open reading frame of MeSWEET17b was 726 bp, encoding 242 amino acids, and located in the plasma membrane. MeSWEET17b had the closer genetic relationship with AtSWEET16 and AtSWEET17, containing 7 transmembrane domains, and belonging to hydrophobic protein.(2)MeSWEET17b mainly transport fructose through the in vitro yeast detection.(3)The results of RT-qPCR showed that the expression trend of MeSWEET17b in stem was basically consistent with in petiole, and the expression was the highest at maturity. The relative expression of MeSWEET17b was relatively low in leaves, and the highest in the expansion stage of tuberous root, while decreased rapidly with the growth of tuberous roots.(4)The ‘KU50' seedlings were subjected to abiotic stress treatments such as high salt(8 g·L^-1 NaCl), drought(100 mmol·L^-1 mannitol), oxidation(10% H₂O₂)and cold(15 ℃ for 24 h, then dropped to 4 ℃ for 24 h). RT-qPCR showed that the relative expressions of MeSWEET17b in leaf and stem had the greatest difference under drought stress. The relative expressions of MeSWEET17b in leaf and fibrous root changed most significantly under salt stress; under oxidation and cold stress, the relative expressions of MeSWEET17b in fibrous root and petiole increased significantly with the extention of treating time. This study provides a theoreticac reference for further studying the function mechanism of sugar transporter protein SWEETs in cassava.

Identification and expression pattern analysis of soybean TrxG gene family

2025/1/7 10:23:14
TrxG(trithorax group)gene family is conserved and widely existed in plants and animals, and plays important roles in regulating gene expression and maintaining normal growth and development of organisms. In order to investigate the distribution of TrxG genes in soybean and its regulatory mechanism in response to abiotic stress of soybean, the physicochemical properties, phylogenetic relationship, composition of domains and genes expression of soybean GmTrxG family were analyzed by bioinformatics method and real-time fluorescence quantitative PCR. The results were as follows:(1)A total of 15 gene GmTrxG family members were identified in soybean, encoding hydrophilic proteins with the length of 982-2 394 aa amino acids.(2)GmTrxG protein members could be divided into three subfamilies, and members of each subfamily contained a SET domain.(3)The expression levels of most GmTrxG genes were highly expressed in stem tips, leaves and flowers of soybean.(4)The genes GmSDG2a/b/c/d were constantly and highly expressed by heat shock and drought. GmSDG2b/c and GmSDG14c were strongly induced by the stress of low temperature, and the genes GmSDG2a/b, GmSDG14c and GmSDG25a/b were up-regulated by salt.(5)Most GmTrxG genes were up-regulated under the treatment of jasmonic acid(JA)and salicylic acid(SA). In conclusion, GmTrxG genes are differentially regulated by different abiotic stresses, and members GmSDG2a/b/c/d, GmSDG14c and GmSDG25a/b may play an important role in soybean response to abiotic stress. The results provide a scientific reference for further exploring the biological function of soybean GmTrxG family.

Cloning, structural and function analysis of MiMYB44L gene in kernels of Macadamia integrifolia

2025/1/7 10:23:14
Macadamia nut(Macadamia integrifolia)is an evergreen nut tree with high economic value. Its kernel is rich in nutrients such as fatty acid and protein, etc. In order to further explore the main regulatory genes related to nutrient formation in M. integrifolia kernels, transcriptomics, gene cloning, fluorescence quantification PCR and bioinformatics techniques were used to screen potential regulatory genes from the kernel transcriptomes of ‘Guire No. 1' and ‘A4', which have significantly different nutrient contents in M. integrifolia kernels. The results were as follows:(1)Transcriptome analysis showed that 1 667 genes were up-regulated and 1 798 genes down-regulated in ‘Guire No. 1' kernel compared with those of ‘A4' kernel; KEGG enrichment analysis showed that the differential genes were mainly in starch and glucose metabolism, amino acid biosynthesis and carbon metabolism.(2)A significant differentially expressed gene gene-LOC122077931 encoding the R2R3-MYB transcription factor MYB44L was discovered. The MiMYB44L gene was cloned in kernels of ‘Guire No. 1' using RACE technology, which was 1 165 bp in length, 999 bp in ORF in length, and encoded 332 amino acids.(3)Bioinformatics analysis confirmed the presence of the SANT domain in the MiMYB44L protein, a hallmark feature of the R2R3-MYB family. The protein lacked both a signal peptide and a transmembrane domain but featured phosphorylation sites.(4)The protein contents in kernels of 10 M. integrifolia varieties were determined. And it was found that the expression of MiMYB44L gene in M. integrifolia varieties with high protein content was significantly higher than that in varieties with low protein content, and the overall correlation coefficient was 0.54, reaching a extremely significant level. The results of this study provide theoretical guidance for in-depth analysis of the regulatory mechanism of MiMYB44L gene in the formation of protein content in M. integrifolia.

Cloning, subcellular localization, and self-activation detection of two NAC transcription factors in response to drought for Camellia oleifera

2025/1/7 10:23:14
Drought stress is a major abiotic stress for the development, yield and quality of Camellia oleifera. NAC transcription factors are widely involved in drought and salt-stress induced signal transduction in plants. To explore the role of NAC transcription factors in the drought stress response of C. oleifera, two-year oil tea seedlings were used as materials. The CDS sequences of CoNAC5 and CoNAC79 were obtained from through TA cloning. Bioinformatics, subcellular localization and self-activation were performed. qRT-PCR was used to determine the tissue specificity of CoNAC5 and CoNAC79 gene expression and the expression PEG and ABA at different treatment times. The results were as follows:(1)Gene structure analysis showed that length of CoNAC5 and CoNAC79 were 1 044 bp and 990 bp, respectively, encoding 348 and 330 amino acids. Their theoretical isoelectric points were 8.86 and 8.57, respectively. The instability coefficients of the proteins were 41.35 and 37.47, respectively. No transmembrane domain was found between the two genes, the highest homology with persimmon and lychee, respectively. Subcellular localization showed that both CoNAC5 and CoNAC79 were located in the nucleus.(2)Yeast self-activation detection analysis revealed that CoNAC5 and CoNAC79 did not have self-activation activity in the full-length proteins and N-terminal domain. However, the C-terminal domain exhibited self-activating activity.(3)The expression of CoNAC5 and CoNAC79 had significant tissue specificity and mainly expressed in roots and kernels; when PEG simulated drought and exogenous ABA treated C. oleifera seedlings, the expression levels of CoNAC5 and CoNAC79 were significantly higher than the control; Furthermore, the expression level of CoNAC79 decreased after 48 h under ABA treatment, and significantly higher than the control under PEG treatment. In summary, it is believed that there may be an inhibitory region at the N-terminal of CoNAC5 and CoNAC79, which hinders the transcription of the full-length sequence; indicating that the two NAC genes in C. oleifera may be probably indirectly involved in drought stress response through pathway of ABA synthesis; CoNAC79 can also directly participate in the drought stress response through other pathways. This study provided a scientific basis for further exploring the role of NAC transcription factors in the response of C. oleifera to drought stress.

Cloning and expression analysis of maize fucosyltransferase gene SPINDLY

2025/1/7 10:23:14
SPINDLY(SPY)is a novel nucleocytoplasmic protein O-fucosyltransferase that regulates target protein activity or stability via O-fucosylation. Previous studies have indicated that the SPY protein regulates plant growth and development by modulating various intracellular processes in Arabidopsis thaliana. Its mediated O-fucosylation plays an important role in maintaining cell homeostasis and regulating plant growth and development, however protein O-fucosylation regulated by SPY in other plants largely remain unknown. Maize(Zea mays )is one of the most important cereals crops for supplying foods, fibers, and fuels to humans. In order to explore the function of maize fucosyltransferase gene(ZmSPY), this study first analyzed the conserved domain, amino acid sequence and physicochemical properties of ZmSPY protein by bioinformatics means, and cloned the gene from maize root tissue to construct the GFP fusion protein expression vector. The subcellular localization of ZmSPY was analyzed, and its response to different hormone treatments(GA, IAA, 6BA, ABA)was determined by exogenous hormone application. And the results were as follows:(1)ZmSPY proteins belong to the TPR and SPY superfamilies, and structural analysis demonstrated that ZmSPY had TPR(Tetratricopeptide repeat)and catalytic domains.(2)Phylogenetic analysis shows that SPYs are highly conserved, and ZmSPY exhibits strong homology to SPY in Sorghum bicolor.(3)Sequence analysis shows that the CDS region of ZmSPY is 2736 bp. Physicochemical analysis indicates that ZmSPY, which contains 911 amino acids and 33 glycosylation sites, is hydrophilic and non-secretory. Its secondary and tertiary structure is largely composed of alpha helix and random coil.(4)The subcellular localization of ZmSPY is predominantly observed in the nucleus.(5)The expression of ZmSPY is induced by phytohormones including GA, IAA, 6BA and ABA, and exhibits various expression patterns. This study provides foundational information on SPY in maize, which could contribute to further investigation of SPY and its effect on O-fucosylation in cereal plants.

Cloning and expression of aerenchyma formation-related gene SmPAD4 in Saussurea medusa

2025/1/7 10:23:14
The aerenchyma is an adaptive structure of Saussurea medusa in response to extreme environments, and its formation is usually accompanied by programmed cell death(PCD). The death of cells and the formation of aerenchyma are typically regulated by the PAD4 gene(Phytoalexin Deficient 4). However, the mechanism by which SmPAD4 regulates the formation of aerenchyma in S. medusa remains unclear. In this study, S. medusa was used as the experimental material, and the gene SmPAD4 related to aerenchyma formation was cloned by homologous cloning and RACE technology, and its sequence, phylogenetic evolution, expression and subcellular localization were analyzed, and its promoter was amplified by hi-TAIL PCR technology to explore its function in environmental adaptation. The results were as follows:(1)The cDNA of SmPAD4 gene was successfully cloned with a total length of 2 047 bp(GenBank accession number OR766038), including an open reading frame of 1 866 bp, encoding 621 amino acids, a molecular formula of C₃₁₆₃H₄₉₀₆N₈₄₈O₉₁₀S₂₆. The protein was an alkaline and hydrophilic unstable protein.(2)Phylogenetic tree analysis showed that SmPAD4 had high similarity with CcPAD4 of Cynara cardunculus.(3)A length of 1 049 bp promoter sequence of SmPAD4 was amplified, which included cis-acting elements such as light response element, hypoxia response element, dry and auxin response elements.(4)Real-time quantitative fluorescence(qRT-PCR)analysis showed that SmPAD4 gene was expressed in root, stem and leaf, and the expression level was the highest in leaf. Under ultraviolet and hypoxia stresses, the expression of SmPAD4 gene was up-regulated in leaf and stem, and down-regulated in root.(5)Subcellular localization showed that SmPAD4 was distributed in the nucleus, cell membrane, and chloroplast. The results show that SmPAD4 gene has a unique protein domain and it responds to hypoxia and ultraviolet environmental stresses, so it plays an important role in the formation of aerenchyma and the response to adversity stress. This study provides theoretical basis for further exploring the role of SmPAD4 gene in the environmental adaptation process of Saussurea medusa.

-->Genetics and Breeding

Comparative analysis on codon usage bias in mitogenome of two species in genus Glycine

Cryopreservation of sterile Pinus massoniana seeds

Effect of simulated drought stress on seed germination and geographical variation of six provenances of Catalpa ovata

Cracking types of common buckwheat pericarp and its effect on early germination traits

Cloning and expression analysis of FmPHV gene in callus of Fraxinus mandshurica

Genome-wide identification and analysis for oil palm WRKY transcription factors

Molecular isolation and tissue expression analysis of alternative oxidase family genes in Liriodendron chinensis

Cloning and expression analysis of CPP transcription factor family LtTCX2 gene in Liriodendron tulipifera

Analysis of ratooning characteristics and quality of distant hybridization buckwheat(Fagopyrum tatari-cymosum)

Techniques for repeated cutting and transplanting of main stem of virus-free healthy sugarcane seedling propagation

Analysis of codon usage bias in the chloroplast genome of Dalbergia odorifera

Growth characteristics of hybrids between Eucalyptus urophylla and Section Exsertaria species

Comparison of expression abundance of genes related to rubber biosynthesis regulation in Hevea brasiliensis

Enhancement of Arabidopsis thaliana seed germination speed by NcEXPA8 gene of Neolamarckia cadamba

Effects of transient infection of FmJAZ1 gene on JA pathway related gene expression in Fraxinus mandshurica

Analysis of genetic relationship and construction of fingerprints in Bougainvillea based on ISSR molecular marker

Condon preference of chloroplast genome of Drynaria

Analysis of codon usage pattern of Eucommia ulmoides

Induction and proliferation of embryogenic callus on improved varieties of fast-growing Pinus elliottii

Screening for in vitro rapid propagation conditions of Adansonia digitata

In vitro regeneration of Osmunda mildei

Analysis of codon usage bias in the chloroplast genome of alfalfa(Medicago sativa)

Codon bias of transcriptomic genes in Cinnamomum camphora

Rapid propagation technology for eggplant rootstock (Solanums torvum)by micro-cuttage

Optimized study for plantlet inducing from cambium stem cell of young stem based on response surface design in Actinidia chinensis

Creating new tobacco germplasm with herbicide- resistance based on oligonucleotide-mediated mutagenesis(OMM)technology

Identification and bioinformatics analysis of WD40 gene family of Meconopsis horridula

Genome-wide identification and expression of millet PAL gene family

Genome sizes and characteristics of cliff plants Opisthopappus taihangensis and O. longilobus on Taihang Mountains

Cloning and fuction analysis of RcCATs and RcSODs genes in Tibet Rhodiola crenulata

Epigenetic diversity of Chinese flowering cabbage revealed by F-MSAP

Genetic diversity of Pinus taiwanensis var. damingshanensis natural populations by SSR markers

EST-SSR marker development and interspecific generality of Albizia odoratissima

Genomic sequence analysis of four culm variants of Moso bamboo(Phyllostachys edulis)on culm

Comparative study on chloroplast genomes of three Magnolia species

Cloning, expression and promoter analysis of LtAGO1 from Liriodendron tulipifera

Cloning and expression analysis of Ml3GT1 in Magnolia liliflora ‘Hongyuanbao'

Codon usage bias in the chloroplast genome of Ventilago leiocarpa

Bioinformatics of grape CBF4 gene and its response to low temperature and potassium silicate

Bioinformatics analysis of WRKY gene family in Physcomitrella patens

Cloning and expression analysis of BeLEA2 gene from Brachymenium exile

Clone and expression analysis of DcSKP1 in Dianthus caryophyllus

Bioinformatics analysis of GeBP transcription factor gene family in Glycin max

Cloning and expression analysis of FtF5H gene from tartary buckwheat(Fagopyrum tataricum)

Screening and transcriptome analysis of EMS-induced cold-tolerant mutants in Hongyang kiwifruit

Sequencing and analysis of full-length transcriptome from Liquidambar formosana leaves in discoloration stage

Cloning and functional analysis of promoter of TcALDH and TcGLIP genes in Tanacetum cinerariifolium

Cloning and expression analysis of FeR2R3-MYB of anthocyanin synthesis-related genes of common buckwheat

Effects of genetic improvement on traits of needle and cone of Cunninghamia lanceolata

Optimization of protoplast preparation conditions of Pleurotus giganteus

Genetic diversity and population structure of Aegilops tauschii accessions based on ISSR method

Identification of resistance to root rot and preliminary establishment of its SSR markers in castor bean

Full-length transcriptome information for Tibetan medicine “Zangyinchen” of original plant Comastoma polycladum

Identification and evolutionary analysis of cotton DUR3 gene

Identification and analysis of TCP transcription factors in response to low nitrogen fertilizer stress in Solanum tuberosum

Discovery and analysis of volatile oil synthesis related genes in Illicium verum based on transcriptome sequencing

Cloning, prokaryotic expression and subcellular localization of PtCHI gene from Pueraria montana var. thomsonii

Development and validity evaluation of Liquidambar formosana EST-SSR primers based on transcriptome sequencing

Functional analysis of RcMsc2 gene in castor(Ricinus communis)

Cloning and expression analysis of the SpLEA1 gene of Selaginella pulvinata under drought stress

Accumulation rule of main nutrients and isoflavones and their relationships in Pueraria lobata var. thomsonii

Physiological differences of different rice materials in response to salt stress at seedling stage

Effects of GABA seed priming on alleviating salt stress and physiological and biochemical changes in Capsicum annuum

Effects of different application modes of phosphorus- solubilizing bacterial fertilizer on seedling root growth and phosphorus nutrition of Pinus massoniana families

Effects of light quality and photoperiod on growth and physiology of Michelia baillonii seedlings

Genetic diversity of six Amorphophallus species in Southwest China based on cpDNA sequences

Transcriptome characteristic analysis and EST-SSR marker development of Capsicum annuum under waterlogging stress

Genetic diversity and structure of wild Rosa roxburghii in Southwest China

Analysis of gene expression characteristics before and after seed germination of Polygonatum cyrtonema

Genetic diversity analysis and comprehensive evaluation of phenotypic characters in 20 germplasm resources of Erythropalum scandens

Phenotypic traits and SSR molecular identification of hybrid progenies of Camellia chekiangoleosa × C. semiserrata

Construction of a mutant library associated with aroma genes in tobacco using CRISPR/Cas9

Bioinformatics and expression analysis of expansin genes GmEXPB5 and GmEXPB7 in soybean

Maintenance and variation of chromosome ploidy under tissue culture conditions of different Camellia germplasms

Influences of polyamines on embryogenic callus proliferation and somatic embryogenesis in Litchi chinensis

Analysis of expression regulation of AP2 gene by pre-miR172 and miR172 in rape

Molecular cloning, location and promoter analysis of NDH dehydrogenase gene from Santalum album

Evaluation and analysis of pine resin components of Pinus elliottii clones

Cloning of HDR gene in Pinus massoniana and Cloningits response to drought and salt stresses