Subcellular Localization Of Potato StCRK1/2 And Preliminary Screening Of Their Interaction Proteins

Potato is one of the world's third largest staple crops,but it is highly susceptible to various diseases during the cultivation process,resulting in reduced yields.Therefore,studying the defense mechanism of potato itself has important theoretical and practical significance for controlling potato diseases.Receptor-like kinases(RLKs)act as conserved upstream signaling molecules that regulate biological processes such as plant development and stress adaptation.Cysteine-rich receptor-like kinase(CRK)is an important class of RLK,which plays an important role in plant disease resistance and cell death.In this study,the CDS sequences of potato StCRK1 and StCRK2 without stop codon were cloned.The potato StCRK1/2-GFP expression vector was constructed by LR reaction in Gateway system,and the expression vector was transformed into Agrobacterium tumefaciens GV3101 for transient expression by infecting N.benthamiana leaves.It was found that both StCRK1 and StCRK2 were localized on the plasma membrane.At the same time,the sequence containing the juxtamembrane and kinase domain in StCRKl/2 was cloned,and the bait vector for yeast two-hybrid was constructed.And a potato cDNA library induced by Phytophthora infestans was constructed.The proteins interacting with StCRK1/2 were screened by yeast two-hybrid technique.After preliminary screening,32 positive clones interacting with StCRK1 and 39 positive clones interacting with StCRK2 were obtained.Our research provides a theoretical basis for further elucidating the function of StCRK1/2 and their roles in disease resistant responses.