Expression Analysis Of Nine Potential Pathogenicity-related Genes And Amidotransferase Gene Knockout And Function Analysis

Banana fusarium wilt is a global devastating disease in banana caused by path-ogenic fungus Fusarium oxysporum f.sp.cubense(Foc)which transmission by the so-il and damage banana vascular bundle.Up to now,we have not quite efficient cont-rol method because of lack of disease-resistant cultibars varieties of banana,unclea-ring of the molpecular pathogenic mechanism and the banana disease-resistant mec-hanism.To illuminate the molecular pathogenic mechanism,provide direction for ba-nana desease resistant breeding and target for the drug development,we selected 9 differences expression proteins pathogenesis related proteins or potential virulence genes from comparative proteomic analysis of Fusarium oxysporum f.sp.cubense race 1 and race 4.The genes were cloned and the expression was analyzed by real time PCR.Fathermore,amidotransferase gene knockout plasmids were build and gene functions were analyzed.The main research results are listed as follows:1.Nine different expression proteins were selected from the comparative prote-omic analysis of Fusarium oxysporum f.sp.cubense race 1 and race 4.We gain ban-ana different expression profiles treated by race 1 and race 4 respectively using rea-1 time PCRAnd four differentially expressed genes were picked out.2.Left and right side gene homologous arms of amidotransferase were amplify-ed and then knockout vector with tag GFP(green fluorescent protein)was construc-ted successfully.3.Ten mutants with GFP(green fluorescent protein)and HPH(hygromycin B phosphotransferase)were got using PEG mediated protoplast transformation gene k-nockout method.4.Got amidotransferase gene knock out strains successfully.5.There are no difference in growth and colonial morphology between gene k-nockout strain and wild strain.6.There have same polycarbafil penetrating power between gene knockout stra-in and wild strain.7.Amidotransferase gene knock out strains have the lower and hydrogen pero-xide tolerability compared with wild strain which suggested that amidotransferase m-ay be play a role in defense the resistance of banana.8.Amidotransferase gene knock out strains have the lower cellulose utilization ability compared with wild strain.9.Amidotransferase gene knockout strain has the higher tolerance to the sorbit-ol and NaCl compared with wild strain.10.Amidotransferase gene knockout strain wasn't able to colonize in banana le-aves while the wild strain was able.11.The amidotransferase knockout strains have the lower pathogenicity compare-ed with wild strain by the pathogenic test.