Functional Identification Of PLC Family Genes Related To The Development Of Lm Female Inflorescence Of Castor Bean

Castor is one of the top ten oil crops in the world and has extremely high economic value.The growth and development of castor inflorescence is one of the factors affecting the yield of castor.The previous research in this laboratory showed that the PLC gene family is related to the inflorescence development of Lm type castor.In order to study the function of the PLC family genes in regulating the inflorescence development of the castor Lm-type female line,this subject took the castor Lm-type female aLmAB2 as the research object,first of all,the bioinformatics analysis and subcellular positioning of the PLC family 6 genes;Then,the expression of 6 genes of the PLC gene family in three types of inflorescences of single castes,standard females,and bisexuals of the castor Lm-type female line aLmAB2 was analyzed by fluorescent quantitative PCR;the 6 genes of the PLC family were in Arabidopsis Heterologous overexpression in mustard,to determine the impact of each gene on the inflorescence development of Arabidopsis;Finally,the relationship between the six genes of the PLC family was analyzed.The test results are as follows:Bioinformatics analysis and subcellular localization analysis showed that the full length of the six genes of the castor PLC family is 1 796-2 163 bp,and the molecular weight of the protein is 62.19-68.95 ku;PLC proteins are all hydrophilic and not Stable protein,the amino acid sequence between them has a high homology of 68.03%;the other 5 proteins except PLC2N are all non-secreted proteins;it is predicted that PLC2N protein is located in mitochondria,and the other 5 proteins are located in other organelles;PLC proteins have no motif structure;in addition to PLC2,PLC6 proteins only contain PLCXc,PLCYc,C2 three domains,the other four proteins contain EF-hand-like,PLCXc,PLCYc,C2 four domains,their The main component of the secondary structure is the ?-helix;the functional domain of the tertiary structure is folded into a dumbbell shape in the spatial structure.The subcellular localization results showed that the castor PLC2N protein localization results were inconsistent with the bioinformatics prediction results,which need to be further verified;the other five proteins are all expressed on the cell membrane,which is consistent with the bioinformatics analysis prediction results.The expression levels of 6 genes of PLC family in castor Lm female line aLmAB2 were analyzed by fluorescence quantitative PCR.The results are as follows:When the expression levels of 6 genes in inflorescences of different types and different development stages were analyzed,PLC2,PLC4,PLC6 genes There are relatively high expression levels in standard female lines;PLC2M,PLC2N,PLC4X2 genes have relatively high expression levels in single female inflorescences;PLC4 genes have relatively low expression levels in single female inflorescences;PLC2,PLC2M,PLC2N,PLC4X2,and PLC6 have relatively low expression levels in inflorescences of both sexes.Six genes of PLC family were heterologously overexpressed in Arabidopsis thaliana.The results showed that:Arabidopsis resistant plants AT-PLCs+(AT-PLC2+,AT-PLC2M+,AT-PLC2N+,AT-PLC4+,AT-PLC4X2+,AT-PLC6+)T3 generation.By PCR detection of AT-PLCS+,6 plants with overexpression of positive genes were obtained,and the results of fluorescent quantitative PCR showed that the expression of 6 genes in AT-PLCs+was higher than that of control wild-type Arabidopsis thaliana.Biological testing of AT-PLCs+showed that the overexpression of PLC family genes can obviously promote the maturation of Arabidopsis thaliana,the growth of lateral moss and the development of flowers and carob,but the basal leaves and stems of Arabidopsis thaliana Leaf development is significantly inhibited.The analysis of the relationship between the 6 genes of the PLC family showed that when the castor PLC2,PLC2M,PLC2N,PLC4,PC4X2,and PLC6 genes were overexpressed in Arabidopsis,the relative expression levels of the Arabidopsis PLC3 gene were all Obviously up-regulated;Based on the analysis of the expression of Arabidopsis PLCs when heterogeneous overexpression of castor PLCs gene and the homology relationship between castor PLCs gene and Arabidopsis PLCs gene,it is speculated that PLC2,PLC2M,PLC2N,PLC4,PLC4X2 and PLC6 genes are in a cooperative relationship.