Screening And Identification Of StCRK1/2 Interacting Proteins In Solanum Tuberosum

In recent years,the potato planting industry has achieved great social and economic benefits.At the same time,due to the particularity of its planting,the degree of trouble caused by diseases and pests is also increasing year by year,which seriously affects the yield and quality of potato.Cysteine rich receptor like kinase(CRK)is an important class of receptor like kinase(RLK),which plays an important role in plant disease resistance and cell death.The team screened 71 positive clones interacting with St CRK1/2 by yeast double hybridization.Six interacting proteins were further screened in this study.Two proteins interacting with St CRK1,St PUB14 and St PUB17,and one protein interacting with St CRK2,St Importin α2.In this study,St Importin α2 was selected for research.the CDS sequence of St Importin α2 was cloned from Solanum tuberosum.Using bioinformatic analysis it has two typical importin α domains in its coding sequence.The binary expression vectors of St Importin α2-G3 and St CRK2-G1 were constructed.The interaction between St Importin α2 and St CRK2 was verified in vivo by using bimolecular fluorescence complementary technique and the interaction was found in the nucleus.Subcellular localization analysis showed that it is located in the nucleus.It was expressed in roots,stems,leaves and flowers.After 4℃ treatment,the expression level of St Importinα2 decreased in most time points.However,its expression level increased significantly after salicylic acid analogue,BTH treatment.This suggests that St Importin α2 may be involved in the disease resistance pathway which is salicylic acid mediated in Solanum tuberosum.The results enrich the research of plant importin family proteins and lay a foundation for elucidating the function of Solanum tuberosum importin α proteins,and the role of St CRK2 in disease and defense response and signal transduction pathway.