Male Sterile Mechanism Of Thermo-Sensitivity Genic Male Sterile PK3-12 In Winter Rape(Brassica Rapa L.)

This paper was mainly study on the main influence factor,flower morphology,physiological and biochemical,endogenous hormone content,two-dimensional gel electrophoresis analysis and differential protein gene expression in the process of fertility transformation about thermo-sensitivity male sterile PK3-12 in Brassica rapa L,and compared with LRA-39 in Brassica rapa L.and NLJ-62 A in Brassica napus L.of cytoplasmic male sterile lines,the main results were as follows:1.PK3-12 was thermos-sensitive genic male sterile,which be fertile under low temperature and be sterile under high temperature.When >0 ? accumulation more than 241.5 ? after appearing bud,PK3-12 can be fertile;when >0 ? accumulation reached 342 ? after appearing bud,PK3-12 can be sterile.2.Fertile period and sterile period of PK3-12 showed the stretched petals,bright color,normal growth of the stigma,length and width of petal,pistil height were no difference,but there is great difference among the stamens,filament and anther length of sterile period were account for 49.8%,71.7% of fertile flower.There was no significant difference among PK3-12?LRA-39 and NLJ-62 A.3.The results of the physiological characteristics show that POD activity and SOD activity of PK3-12 in sterile period were higher than that of fertile period,while the CAT activity showed the opposite regular.During the sterile period,calyx cracking stage of POD activity than that of young bud stage increased 32.2%,flowering stage of POD activity than that of calyx cracking stage increased 26.3%,during the sterile period,calyx cracking stage of CAT activity than that of young bud stage reduced by 45.7%,flowering stage of CAT activity than that of calyx cracking stage reduced by 31.7%.Changes regular of POD activity and CAT activity of PK3-12 were consistent with LRA-39 and NLJ-62 A,while the changes regular of SOD activity of PK3-12 was opposite with that of LRA-39 and NLJ-62 A.The content of soluble protein in three male sterile lines were maintained at a low level and the content of soluble protein of PK3-12 in sterile flower changes more dramatically.4.The results of study on endogenous hormone content of PK3-12 showed that ZR and GA3 content of PK3-12 had remained at a low level with the development of flower organ in sterile period,while ABA content always remaining at a higher level;ZR,GA3 content of PK3-12 were all lower than LRA-39 and NLJ-62 A in the bud stage,calyx cracking stage and flowering stage.5.The results of differential protein identification by mass spectrometry showed that there were Ru BP carboxylase/oxygenase,phosphoglycerate kinase,glutathione transferase/lyase appeared in PK3-12 fertile and sterile period in different proteins,LRB-26 and LRA-39 differential proteins,NLJ-58 and NLJ-62 A differential proteins.however most differential proteins also showed species specificity: Specific known proteins of PK3-12 were heat stress protein,membrane associated protein,succinyl coenzyme A,UMP-c MP kinase;specific known protein of LRB-26 and LRA-39 were starch synthase,aspartic protease,chlorophyll a/b binding protein,deoxycytidine deaminase,ribosomal protein;specific known protein of NLJ-58 and NLJ-62 A were the S-glutamine synthetase,S-adenosylmethionine synthetase,malate dehydrogenase,proteasome subunit beta,peptidyl prolyl CIS trans isomerase protein.6.Differential protein GO functional annotation showed that differential protein of PK3-12 in fertile and sterile period mainly concentrated metabolic reactions,metabolic process,response to stimulus,single-organism process in the biological process;concentrated binding and catalytic activity in molecular function;concentrated membrane,organelle,cell part in cell component,which were consistent with GO concentration analysis results of LRB-26 and LRA-39 differential protein,NLJ-58 protein and NLJ-62 A differential protein,but the percentage of the same GO term in three kinds of differential proteins were different.The KEGG pathway analysis of differential proteins shows that the three kinds of differential proteins were mainly involved in glycolysis,amino acid biosynthesis,photosynthetic biological carbon fixation and other pathways.7.The results showed that relative expression of RCA gene(common differential protein gene)in fertile period of PK3-12 was significantly lower than the sterile period in each stage,while the other two cytoplasmic male sterile materials showed the opposite regular,and the degree of deviation of relative expression of RCA gene in PK3-12 between male sterile period and fertile period was more severely than two kinds of cytoplasmic male sterile lines.The relative abundance of differentially expressed proteins was used to verify the abundance of protein expression,we find that not all of the relative expression of differential expressed genes was consistent with the abundance of protein expression.