Preliminary Study On Establishing Stable Genetic Transformation System Of Wheat Stripe Rust

Stripe rust,caused by Puccinia striiformis f.sp.tritici(Pst),is a disease on leaf spread by gas and a serious fungal disease of wheat worldwide.Pst,belonging to Basidiomycotina,is a strict obligate biotophic fungus and can not be cultured on artificial culture medium.As the sequencing of Pst genome completes,the biology of Pst enters into functional genomics era,so that the tools for studing functional genomics will be needed largely.However,genetic transformation technique is a strong tool for detaching and analyzing gene function.Therefore,it is urgent to establish a stable genetic transformation system of Pst for functional genomics research in depth.In this study,we select the promoter of ?-tubulin gene which is stably expressing in wheat stripe rust as transformation vectors' promoter.The mCherry act as a reporter marker,because it is a monomer molecules and stable fluorescent label.We select ?-tubulin and Bar gene as screening markers and construct a series of transformation vectors.Then,we use detached leaf in vitro to determine a suitable benomyl concentration for screening transformed strain.In this study,we introduced DNA into urediospores by using the Agrobacterium-mediated transformation system and biolistics-mediated transformation system.The vectors are constructed for biolistics and Agrobacterium-mediated transformationsystem,by inserting Pst ?-tubulin promoter for mCherry,?-tubulin and Bar genes fragment into pBluescript KS ? or PBin20.For chemical screening,benomyl was selected.The concentration 75?g / ?L was determined for Pst transformats screening.So far,we have tried to insert DNA into urediospores by biolistics and gotten a certain collections of putative transformants of Pst.