Screening, Identification And Functional Analysis Of Genes Interaction With Clg2p From Curvularia Lunata

Maize is a very important grain crop in China even in the world, Curvularia leaf spot ofmaize caused by [Curvularia lunata（Walker） Boed.] threats the yield of maize seriously. So thepathogenic mechanism research of this plant pathogen can provide theoretical basis in preventionand control. Clg2p coding Ras homology protein has been cloned in our research group,comparing the mutant ΔClg2p deletion of Clg2p with the wild Curvularia lunata DQ-1,indicated that the conidiospore yield of ΔClg2p reduced, conidiospore morphology,appressorium morphology and pathogenicity aspects of ΔClg2p had some changes to a certainextent, these phenomenon indicated that Clg2p contributes to biological characteristics andpathogenicity in Curvularia lunata, but until now the regulation mechanism of Clg2p stillunknown. Clg2p was used as bait in this study and7proteins interacting with Clg2p werescreened out from cDNA library of Curvularia lunata; after sequencing anylasis, function of Clfand CIUase in biological character and pathogenicity were verified, the results of this study areas follows:1. Clg2p as bait,7positive clones were screened out by yeast two hybrid, they wereUricase(CIUase)、 Raf protein kinase (Clf)、 Cell wall-associated hydrolase、 Zincfingertranscription factor ace1、Acetyl-coenzyme A synthetase and two hypothetical proteins；2. Yeast two hybrid and BiFC assays further confirmed that Clg2p interact with Clf andCIUase;3. Clf and CIUase deletion homologous fragments were constructed by using Double-jointPCR;59Clf deletion mutants and63CIUase deletion mutants could grow on regenerationmedium with hygromycin,1positive Clf deletion (ΔClf) and1positive CIUase deletion(ΔCIUase) were screened out from59Clf hygromycin resisitant mutants and63CIUasehygromycin resistant mutants;4. Comparing pathogenicity and morphology between wild type Curvularia lunata (DQ-1)、ΔClf and ΔCIUase, the results indicated that ΔClf can’t produce conidiospore, pathogenicity ofΔClf weaken; conidiospore morphology of ΔCIUase had changed, most of conidiosporeproduced by ΔCIUase turned into straight instead of falcate, pathogenicity of ΔCIUase weakenedtoo. the result confirmed that Clf regulats pathogenicity and conidiospore production; ClUaseregulated conidiospore morphology and pathogenicity.