Functions Of Stress Related Genes Mediated By ClpA ATPase In Xanthomonas Campestris Pv.Malvacearum

Cotton angular leaf spot is a cotton disease caused by Xanthomonas campestris pv.malvacearum(Xcm).It is less harmful and is a minor diseases in the history.Studies on the cotton angular leaf spot is less than other diseases.In recent years,due to the spreading of Sea Island Cotton varieties and the changes of climatic conditions,the incidence of the cotton angular leaf spot increased these years.Therefore,the study of the pathogenesis of Xcm has great value for the development of effective prevention and control measures.ClpA is a conserved bacterial protein quality control factor,which can promote the degradation of abnormal proteins in cells by hydrolysis of ATP,and the role of ClpA in Xcm has not been reported.In previous studies,we demonstrated that ClpA is involved in the pathogenicity of Xcm to cotton,but the pathogenic factors and metabolic pathways regulated by ClpA remain to be elucidated.In this study,we investigated the regulatory effects of ClpA on Xcm by detecting the expression of 5 differentially expressed proteins in the wild-type and AclpA mutant of the wild type and the phenotype of the defective strains.The partial amino acid sequences of the differentially expressed proteins of Xcm wild type and clpA mutant were determined by the method of bioinformatics and compared with the previous proteomic methods,and 5 differentially expressed proteins were identified.They are 2-oxoglutarate semialdehyde dehydrogenase(ketoglutarate semialdehyde,dehydrogenase,KsdD),penicillin binding proteins(penicillin-binding,protein,PbP),two carboxypeptidase(dipeptidyl peptidase,DpP),isoAspartate protein carboxyl methyltransferase(L-isoaspartate protein,carboxylmethyltransferase,IpcT)and bacterioferritin co migration of protein(bacterioferritin comigratory protein,BcP).In order to reveal the biological function of these 5 proteins in bacteria Xcm in the role of the ClpA protein in the regulation of Xcm pathogenicity,gene expression sequence flanking the protein encoding by difference,connected with the sucrose mortality characteristics pK18mobSacB,the gene knockout vector was constructed,and the 5 derivative mutants△ksdD,△pbP,△dpP,△ipcT and AbcP were obtained by homologous recombination.Determination of the five its derivative mutants respectively,in biofilm formation,movement ability,salt resistance,heat resistance,competition,colonization and other aspects of the phenotype.The results showed us that:compared to the wild type,△pbP biofilm formation ability decreased significantly,athletic ability is also weakened,the tolerance of salt ions was also reduced,the pathogenicity of ApbP is far less than that of wild type,speculated that pbP participates in the regulation of multiple genes in Xcm.In order to further prove the role of gene ApbP under adversity stress in Xcm,complementation strain by homologous recombination,phenotypic differences between wild type and mutant and complementary strain in the biofilm,salt movement,etc.After the discovery of gene complementation,we have a certain degree of recovery for the tolerance of salt ions,exercise capacity also increased.Through further study of the function of pbP and its role in Xcm,it is expected to reveal the function of stress related genes mediated by ClpA ATPase.