| Cuticles acts as a barrier to protect plants adapt to the stress,which is closely related with the anti-adversity of the plant,cutin is the main element of the cuticles,which is also the basic structure of cuticles layer,the research on cutin helps to clarify the molecular mechanism of the formation and regulation of cuticles,which also lays the foundation for the research of the function of the cuticles.In this paper,we optimized the cutin extraction method,by Gas chromatography-mass spectrometry(GC-MS)technical analysis to identify cutin content and composition,compard the two grown rape cutin content composition differences and rapeseed different organizations(stem,leaf,silique)cutin content composition differences,the different degrees of drought stress on eight drought resistance there is significant differences of the effect of rapeseed varieties leaf cutin,we used the RNA-Seq sequencing technology testing different(root,stem,leaf,flower and silique,seed)and we found the of gene expression under drought stress,we selected some cutin genes by the analysis of the related gene expression level.we identified 398 rape strains of cutin content and composition.The main results are as follows.We found the pyrolysis liquid of methanol-sulfuric acid-chloroform effect is the best way,separation and identify the cutin of monomer composition,most cutin monomer amount higher than the other three methods,which is suitable for cutin extraction.The rape leaf cutin composition is more comprehensive in filed environment,and most of the relative content of cutin composition is higher than that in greenhouse environment.We analyzed the cutin content in silique,stem and leaf on double 11 rape,a total of 81 cutin monomers.silique,stem and leaves were found to have the highest relative contents of cutin is 2-hydroxy-Tetracosanoic acid.the relative content of 5 cutin components,Tetradecanoic acid?14-methyl-pentadecanoic acid?hexadecanoic acid? 2-hydroxy-Tetracosanoic acid?20-methyl-2-hydroxy-docosanoic acid were significant different in silique,stem and leaves.RNA-Seq sequencing technology was used to analyze the gene expression in different tissues of double 11 rapeseed,The expression of the genes in 118 cutin was analyzed.Six genes were not expressed in different tissues.BnaA05g09070 D is only expressed specifically in seeds.BnaC05g04230 D is only expressed specifically in flowers.BnaCnng13910 D has the highest expression in the root and the leaves,and the expression of BnaA04g20340 D is the highest in the flowers.The expression of BnaC04g51420 D is the highest in the stem,and the expression of BnaA02g32750 D is the highest in the silique and the seed.The cutin content of eight rape strains in different drought conditions was analyzed.Compared with the control group,with the increase of drought stress,the total cutin decreased.Compared with the control group,the relative content of 27 cutin in 8 cultivar was significantly difference.The expression of the 118 cutin genes related to Cutin synthesis was analyzed at different time periods(0h,1h,8h)during the drought stress,.there are 108 different genes,Seven candidate genes were significantly different,and the expression of four candidate genes increased significantly,The expression of three candidate genes decreased significantly.We have analyzed the cutin contents and components of 398 strains of rapeseed,and the next step is to make GWAS analysis of the cutin content.and to locate the locus on some chromosomes related to Cutin synthesis. |
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