Research On The Screening,Cloning And Function Of Rice Blast Fungus Effector Proteins

Background and Objective:Rice blast fungus effector protein (EP) play an important role in the pathogenesis of Plant Pathology, so there are many majority scholars pay more and more attention on it.The study on EP can reveal the molecular interaction between rice blast fungus and rice. Even help the rice occurrence of blast. At present, it is difficult to explore EP and study its functions.In this study, we hope through prediction of rice blast fungus genome analysis and transcriptome analysis of high-throughput sequencing after 36 hours infection,combined with the specificity of gene expression to screen out some important EPs and to study their functions,We also hope to provide new routes for exploring the cloning of pathogen effectors.Methods:(1)Through bioinformatics,we analysis rice blast the predicted genes of fungus genome and analysis high-throughput sequencing of total RNA after ROR1 infected Nipponbare rice leaves 36 hours,meantime,we combine with the expression of the gene-specific validation (RTPCR),the ultimate goal is to screen some EPs that have disease-related.(2)The EP will be filtered out into the rice over-expression vectorâ–³pCambia1305.2.Then,through the rice transformation to obtain transgenic lines of rice.(3)2-3018 was found has BIR domain,and then through constructing expression vectors pCB1532,knockout vector (â–³T),recombinant promoter vector pCSN43 of the rice blast fungus,and series of pvx transient expression vectors of tobacco,we want to study its function.Results:(1)Through genome analysis and high-throughput sequencing data analysis,we successfully obtained 45 candidates EPs, and eventually to identify 12 EPs to research including the 2-3018, and made sure that the above methods is feasible at discovering and cloning of EPs. (2)Constructed expression vector of EPs and transformation of rice and obtained transgenic rice plants.(3)Constructed several carriers of 2-3018, made the foundation for further study of its function.Conclusion:(1)Successfully screened out a part of the EPs,and provided a new way of exploring and cloning of pathogen effectors. (2)Successfully constructed rice expression vector pCambia1305.2-EPs, and through rice transformation,obtained transgenic lines of rice. (3)Successfully unearthed the BIR domain of 2-3018,and build its overexpressive vector pCB1532-3018,knockout vectorâ–³TKF2-3018 andâ–³TKFbir, recombinant promoter vector pCSN43-3018F1 and pCSN43-3018F2 of rice blast fungus,and transient expression vectors PVX-3018,PVX-C93,PVX-C96,PVX-H109,PVX-H114 of tobacco.