Function Analysis Of VdWRKY70 Transcription Factor Disease Resistance From Vitis Davidii 0943

C.diplodilla is one of the four major diseases of grape,so it has become an important research direction of grape resistance to C.diplodilla.As one of the origin centers of grape plants in the world,China has many valuable wild grape resources.Some of the varieties were resistant to C.diplodilla,especially ‘Vitis dividii 0943’ has strong ability to resist C.diplodilla.In the early stage of the study,it was found that Chinese wild grape ‘Vitis dividii 0943’ had strong resistance to C.diplodilla,and that Vd WRKY49 and Vd WRKY53 played an important role in the resistance to C.diplodilla.This study research the function of VdWRKY70 transcription factor with resistant to C.diplodilla by contrast with Manicure Finger.The main research results are as follows:1.Through phenotypic observation of C.diplodilla and Trypan blue staining at different time after inoculation in VAC and VIM,it was found that the ability of resistance to C.diplodilla was stronger in VAC.By combining the content change of SA,JA with the relative expression of WRKY70 and related disease resistance gene NPR1/PR1,it was found that the change trend of VdNPR1 and VdPR1 was consistent with the change trend of SA content.Therefore,it is speculated that Vd WRKY70 regulates the resistance to C.diplodilla of VAC mainly through SA pathway.2.Through the localization and nuclear localization of Arabidopsis thaliana protoplasts,it was found that Vd WRKY70 had nuclear localization signal and was located in the nucleus of Arabidopsis thaliana.3.The culture of Arabidopsis thaliana seedlings transferred with Vd WRKY70,wild type Arabidopsis thaliana as control,which inculated with C.diplodilla,G.cichoracearμm and Pst DC3000.After inoculation,We observe the phenotypes of Col-0 and Col-PGW70 by a series of means,such as phenotypic observation,trypan blue staining and statistical analysis of quantitative characters.It was found that Col-PGW70 had stronger disease resistance than Col-0.We combined the change of SA content in Col-0 and Col-PGW70 with the change of relative expression of Vd WRKY70,AtNPR1 and AtPR1.It showed that the trend change of SA and relative expression of AtNPR1 and AtPR1 were basically consistent.4.By cluster analysis,it was found that the VdWRKY70 protein sequence was clustered with Fc WRKY70,CaWRKY70,At WRKY70,AtWRKY54 and ClWRKY70,indicating that they also had some similarity in function.5.The promoter sequences of NPR1 and PR1 in Vitis dividii 0943 and Beautiful finger were obtained by homologous cloning.The results of sequence alignment and cis-acting element analysis showed that VdPR1 and VvPR1 promoter sequences are different,VdPR1 sequence is 26 bp more than the VvPR1 sequence,and there are cis-acting elements related to SA pathway.Therefore,we speculate that Vd WRKY70 transcription factors may combine with VdPR1 promoter in SA pathway.