Castor Different Types Of Inflorescence Differences Studied By DNA Methylation And Proteomics

Castor is one of the ten oil crops of dicotyledon,which is the only vegetable oil that can replace oil.Increasing the yield of castor per plant at this stage is one of the methods to improve the yield of castor.Castor’s inflorescence type directly affects the yield of castor.The female line of Lm female was obtained by 60Co-C ray treatment from Tongliao Academy of Agricultural Sciences.It is an important castor breeding material in Inner Mongolia,including three species of per female,marked female and amphoteric.Therefore,in this study,the inflorescence axis of Lm female line,per female,marked female and amphoteric at different developmental stages was studied.DNA methylation and proteomics differences were studied to find the genes and proteins related to the development of castor inflorescence.This study laid the foundation for revealing the molecular mechanisms underlying the development of castor inflorescence from gene levels and protein levels.The level and pattern of inflorescence methylation of per female,marked female and amphoteric plants at different developmental stages were analyzed by MSAP technique.The results showed that the total methylation rate,semi-methylation rate and total methylation rate were decreased during the 4-leaf stage to the 5-leaf stage.And the total methylation rate,hemi-methylation rate and total methylation rate were increased during the 5-leaf stage to the main spike and two branches.The methylation rate was the lowest at 4-leaf stages and 5-leaf stages.Sequencing successfully recovered of 256 differential bands of which 28 have functional loci and are homologous to known functional genes.28 sequence analysis showed that C11,C16,C18,C28 may be involved in the regulation of the development of castor mosaic.C11,C16,C18,C28 were tested for bisulfite sequencing.The results showed that C11 had methylation differences only in the 4-leaf stage and the two branch of the per female inflorescence.C16 had methylation differences in the four-stage marked female inflorescence;C18 had methylation differences in the four-stage marked amphoteric inflorescence.C28 in the 4-leaf stage,5-leaf stage,the main spike period of per female inflorescence in the presence of methylation differences.C11,C16,C18 and C28 genes were analyzed by fluorescence quantitative PCR,indicating that gene hypermethylation inhibited gene expression,low methylation promoted gene expression.These results indicate the involvement of DNA methylation in the process of formation of castor.Two-dimensional electrophoresis was used to analyze the inflorescence of per female,marked female and amphoteric at different developmental stages of castor.The result of two-dimensional electrophoresis showed that the phenol extraction method was more effective than TCA acetone precipitation extraction method.Image Master 2D Platinum Software Version 7.0 software for protein profiling,the 93 differences were particularly significant for the recovery of protein spots.84 protein spots MS analysis was successful,of which 80 successfully searched the protein library.The 80 differential proteins were classified into 9 categories by function(substance metabolism related proteins,energy metabolism related proteins,amino acid metabolism and protein biosynthesis related proteins,signal transduction related proteins,photosynthesis related proteins,development of related proteins,transport related proteins,other proteins,unknown protein).The analysis of 80 proteins by thermal analysis showed that in the same development period,different types of inflorescence of different proteins.The difference between per female and mark female is far less than that between per female and amphoteric,between mark female and amphoteric.The same type of inflorescence in different developmental stages of the differential protein,there were differences between the 4 leaf period,the 5 leaf period,the main spike period and the two branches period.But there was a significant difference 4 leaf and 5 leaf period between main spike and two branches period.Analysis of 80 proteins showed that M15,N3,L9 and N7 may be involved in the development of castor.Integrated DNA methylation and proteomics results show the differential gene fragment C18 and differential protein M15 were involved in the phosphatidylinositol signaling system,which affected the development of the flowering of the castor by affecting the morphology of the vacuoles and the development of pollen tubes.