Screening And Identification Of Effectors RxLR(Nudix)and Rx LR548 Interaction Proteins In Phytophthora Capsici

Phytophthora capsici is one of the main diseases of Capsicum annuum,which is distributed all over the world and can harm 78 crops in 18 families including leguminous,cucurbitaceae and Solanaceae,resulting in reduced yield and severe economic loss of Capsicum annuum.In recent years,it has been found that Phytophthora capsici secretes a large number of effectors,which plays a vital role in the invasion of pathogenic.Plants have a large number of target proteins to help pathogens infect or inhibit the invasion of pathogens.Therefore,it is of great scientific significance to carry out in-depth research on the isolation and identification of Phytophthora capsici and its target protein in the host.In this study,the target protein of the effectors in the host was used as the research object,and the protein interacting with it was screened by yeast two-hybrid method,and the effectors was further determined by the combination of bimolecular fluorescence complementation(BIFC)and co-immunoprecipitation(Co-IP).The interaction between the effectors and the target protein.The main findings obtained are as follows:1.Using yeast two-hybrid technique,RxLR548 screened five suspected interacting proteins.After one-to-one rotation verification,three interacting proteins were identified as bZIP transcription factor 17,protease inhibitor(PI),and glyceraldehyde-3-phosphate dehydrogenase A(GAPD).2.RxLR101(Nudix)is a key functional domain of RxLR101.RxLR101(Nudix)screened six suspected interacting proteins,and confirmed the 2-hydroxyisoflavone dehydratase(HID)as the target protein of the gene through one-to-one rotation verification.3.RxLR101(Nudix)and 2-hydroxyisoflavone dehydratase-like have been shown to interact by bimolecular fluorescence complementation assays,and the interaction positions are nuclear and cytoplasmic.4.In order to further determine the interaction between RxLR101(Nudix)and2-hydroxyisoflavone dehydratase-like,co-immunoprecipitation experiments were carried out.The results also showed that 2-hydroxyisoflavone dehydratase-like is indeed RxLR101(Nudix)in Capsicum annuum Target protein.Through the above studies,we found that the suspected target proteins screened by the above RxLR effectors are mostly related to the disease resistance and anti-reverse pathways and signal transduction.It is speculated that the above RxLR effectors may cause related disease-resistant immune protein production to prevent the invasion of pathogens in plants when infecting the host.The results of this thesis provide a basis for further research on the mechanism of interaction between RxLR effectors and interaction proteins,and provide a theoretical basis for the prevention and control of Phytophthora capsici and the mechanism of plant disease resistance.