Clone And Functional Analysis Of S-phase Kinase Associated Protein Gene PsSkp1 In Phytophthora Sojae

The Soybean Root Rot is one kind of ruinous earth passes on plant disease,causes by Phytophthora sojae.The spreading of Phytophthora sojae is mainly through the mycelium and the zoospores.If we can destroy the growth of mycelium and the production of zoospores,it can effectively control the disease.In this paper,the Skp1 is a member of ubiquitin ligase SCF complex family(Skp1-Cul1-F-box),the protein can regulate the degradation of proteins in the organism,and participate in many aspects of biological development through the degradation of multiple cell cycle regulatory proteins.In this study,we constructed the Skp1 gene silencing expression vector pTORSkp1-GFP in Phytophthora sojae by RAN interference technology.Using the method of protoplast mediated transformation,the silencing vector was introduced into the wild-type Phytophthora sojae 6497,so that the target gene was transiently expressed.Through the qRT-PCR experiment,we selected two strains which had low expression of Skp1 gene in the mutant strain,named L1 and L2.Phenotypic analysis of the mutant was carried out by stress,infection of hyphae,induction and pathogenicity of conidia and electron microscopy.The growth rate was measured by the experiment on silent mutation strains,found that the growth rate is relatively slower than silence mutant;and analyzed its antioxidant capacity and salt tolerance were weakened;I induced strains of sporangia and zoospores,found the number of silence mutant zoospores decreased,as the original strain 20%-30%;Though measuring the pathogenicity of mycelial growth rate of the mutant,I found silence deformity having more branches,the top is thick,early aging and dying,soybean infection and pathogenicity were lower;zoospores infection and pathogenicity were low on soybean yellow flower seedlings.