Subjects: Biology >> Botany >> Applied botany submitted time 2023-12-24 Cooperative journals: 《广西植物》
Abstract： The secondary laticifer is the position for synthesis and storage of natural rubber (NR), which is differentiated from the vascular cambium of bark in stem of rubber trees (Hevea brasiliensis). The quantity of secondary laticifer is depend 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. So, 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 is successful and constructed by the technology of gateway. The capacity of the primary library is 6.3 × 106, the total number of clones is 1.2 × 107, and the capacity of secondary library is 7.7 × 106, the total number of clones is 1.5 × 107, the recombination rate of the libraries is 100%. (2) The average length of inserted fragments is 1.1 kb and 1.2 kb in primary and secondary library, respectively. The bait vector of pGBKT7-HbHDA6 is constructed and confirmed no self-activation activity. (3) The cDNA library is screened with the pGBKT7-HbHDA6 bait vector, found 22 proteins interacting with HbHDA6, 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.
Subjects: Biology >> Botany >> Applied botany submitted time 2019-08-26 Cooperative journals: 《广西植物》
Abstract： 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, it is known that, 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 is yet to be elucidated. In the present study, the expression abundance differences of 15 rubber biosynthesis regulatory genes HbCOI1, HbJAZ1, HbJAZ2, HbJAZ3, HbMYC1, HbMYC2, HbMYC3, HbMYC4, HbMYC5, HbGAPDH, HbHMGR1, HbSRPP, HbREF, HbHRT1, HbHRT2, and 2 common internal reference genes Hb18S, HbACTIN1 in 10 rubber tree germplasms latex following tapping them with S/2D d/3 tapping system were compared. The expression abundance of HbACTIN1 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 showed that 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, respectively. Correspondingly, 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 percent. Overall, at the population level, the transcription abundance of the 16 genes from high to low were 18S＞SRPP＞HMGR1＞REF＞MYC2/HRT1＞COI1＞MYC1/MYC4＞GAPDH/JAZ1/MYC5＞JAZ2＞HRT2/MYC3/JAZ3, correspondingly, the average abundance were 28382.26, 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 HbACTIN1 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 to understanding the function of genes in a more comprehensive way.
Subjects: Biology >> Botany >> Applied botany submitted time 2019-08-26 Cooperative journals: 《广西植物》
Abstract： Tapping-enhanced rubber biosynthesis is closely related to the activation of jasmonic acid signaling in laticifer cells of rubber tree. The expression level of genes related to jasmonic acid signaling pathway and rubber biosynthesis were both positively correlated with the dry rubber yield. However, the exact relationship between the expression of gengs involved in jasmonic acid signaling and rubber biosynthesis is yet to be elucidated. In the present study, qPCR was used to analyze the expression of 9 jasmonic acid signaling genes, HbCOI1, HbJAZ1, HbJAZ2, HbJAZ3, HbMYC1, HbMYC2, HbMYC3, HbMYC4, HbMYC5 and 6 rubber biosynthesis genes, HbHRT2, HbSRPP, HbREF, HbHMGR1, HbHRT1, HbGAPDH, in laticifer cells of 5 Wichham germplasms and 5 1981’IRRDB germplasms following tapping them with S/2D d/3 tapping system, Hb18S was used as internal reference gene. The correlation between these 15 genes was analyzed by Pearson correlation coefficient. The bivariate correlation coefficient (r12) and partial correlation coefficient (r12.3) of 105 gene pairs were obtained with the mean ± standard deviation were 0.486 ± 0.220 and 0.304 ± 0.211, respectively. Among them, 63 gene pairs (60%) with r12 in the same direction as r12.3 and 42 genes pairs (40%) in the opposite direction; In the degree of correlation, 23 gene pairs (21.905%) with | r12 | less than | r12.3 |, and 82 gene pairs (78.095%) with | r12 | greater than | r12.3 |. There 76 gene pairs (72.38%) bivariate correlation coefficient (r12) were significant correlation at P＜0.05 level and 59 gene pairs (56.19%) at P＜0.01 level. In contrast, there less partial correlation coefficient (r12.3) were significant correlation at P＜0.05 level (21pairs, 20%) and P＜0.01 level (16 pairs, 15.24%). These results suggest that the expression of genes involved in these two pathways were related to each other. And provides a further evidence for the assumption that “expression correlation of functional related genes”, which is widely adopted in gene expression profile analysis. It can be used for excavating, screening and predicting unknown genes related to rubber biosynthesis, as well as provides a theoretical basis for studying the molecular regulation mechanism of rubber yield formation in Hevea brasiliensis Muell. Arg.
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