Identification And Preliminary Function Analysis Of Candidate Effector Gene Cg4LysM From Colletotrichum Gloeosporioides Hevean Brasiliensis

Anthracnose is one of the important fungal disease of rubber tree,which is mainly caused by C.gloeosporioides(Colletotrichum gloeosporioides),rubber can infect leaves,petioles,the yield of rubber caused a huge impact.Effector protein is one of the major causative factors of pathogenic fungi,due to the current limited research rubber tree anthrax LysM type of effector protein,so in order to control the rubber anthrax,this experiment chose effector proteins having a conserved domain structure Cg4LysM LysM as a candidate effector proteins,isolation and identification of them,then a preliminary analysis of its function.In this study,the Cg4LysM gene was cloned according to PT-PCR.According to bioinformatic analysis shows that the gene contains 1983bp bp open reading frame(ORF),encoding 660 amino acids.The protein contains four conserved LysM domains,amino acids 1-20 of protein is typical of a signal peptide sequence,without any transmembrane domains,presumed to be extracellular secreted protein.Comparative analysis by BLAST alignments and software DNAMAN its homology found anthrax spores gum rubber Cg4LysM strawberry gloeosporioides(Colletotrichum gloeosporioides Nara gc5),Houttuynia anthracnose(Colletotrichum gloeosporioides Cg-14)sorghum anthracnose(Colletotrichum sublineola),plaster sample microspore bacteria(Microsporum gypseum CBS),two grapes mold(Stachybotrys chartarum)protein having high homology,homology were 97%,95%,55%,59%,59%.To further investigate the function of effector proteins Cg4LysM constructed knockout vector designated pCB1532-ToxA-?Cg4LysM.By the principle of homologous recombination,protoplasts obtained knockout mutant ACg4LysM.The results showed that:the growth rate of mutant Ming lower than the wild-type growth rate,aerial mycelium and undifferentiated wild type conidia compared with wild type,mutant glucose,sucrose,galactose utilization rate is higher.By pathogenic analysis shows that the incidence of the mutant decreases the incidence is lower than the wild type,but pathogenic mutants have not lost,knockout mutant strain mechanical force is not lost.Oxidative stress tolerance of the wild-type non-discriminatory.It follows that the gene is a non-pathogenic factor Cg4LysM rubber gum anthrax spores,but the gene may be involved in the regulation of pathogenicity of the strength.Hevea rubber trees anthrax gene Cg4LysM no significant pathogenicity,but involved gloeosporioides spores and hyphae growth regulation.