Cloning And Disease Resistance Of Sucrose Transporter Gene In Grapevine

Vitis vinifera L.is a widely cultivated economic fruit tree in the world.With the implementation of rural revitalization in China,its economic value has become increasingly prominent.Grape is susceptible to fungal infection in the production process,resulting in grape diseases,reducing fruit quality and yield,affecting economic benefits.Botryosphaeria dieback is one of the most important diseases,of which Lasiodiplodia theobromae is the most virulent.Glucose transporter is the key to regulate the immune function of plants against pathogens,but so far,the mechanism of grape immune has not been reported.The aim of this study was to reveal the role of sucrose transporters in the interaction between host grapes and L.theobromae,and to provide new ideas and basis for the prevention and control of grape canker,especially for the prevention and control of L.theobromae,and the cultivation of resistant varieties.In this study,we explored the functions of sucrose transporter genes in disease resistance and the ways of participating in grape disease resistance regulatory networks.The following research results have been achieved:1.Cloning of SUC gene from ’ Thompson Seedless ’.Bioinformatics analysis showed that this family of proteins had 12 transmembrane regions,and the helix structure model was basically consistent.It had close genetic relationship with other sucrose transporters.The subcellular localization was on the cell membrane,which conformed to the properties of sucrose transporters.2.Immune-related factors induce SUC gene expression.Four different hormones(SA,Me JA,ACC,ABA)and two elicitors(chitin,flg22)were carried out on the tissue culture seedlings of ’Thompson Seedless ’ grapes,and the inoculation treatment of ’ Summer Black’ grape branches was carried out.It was found that the three genes had different degrees of response to the above immune response-related factors,indicating that they were involved in the relevant response pathways of grape to the infection of L.theobromae.Through the comparison of gene expression results,it can be seen that the expression peak of VvSUC12 gene under various treatments was significantly higher than that of the other two genes.It is inferred that the gene may have a higher correlation with the disease resistance pathway of grape.However,VvSUC27 shows great differences,which may have functions different from those of VvSUC11 and VvSUC12.3.Functional verification of VvSUC12 gene.The over-expression of VvSUC12 gene in tobacco and grape callus was obtained by transgenic technology.Through the inoculation observation,it was found that grape callus showed resistance to L.theobromae,indicating that VvSUC12 gene could improve the disease resistance of plants.4.Screening of Dof transcription factors related to the upstream of VvSUC gene.Six Dof transcription factors that might interact with the VvSUC12 gene were screened out by yeast onehybrid assay.Yeast self-activation verified that VvDof19 and VvDof22 had transcriptional activation activities.Post inoculation showed that the expression patterns of VvDof19 and VvSUC12 were similar.GUS activity test proved that VvDof19 could activate the expression of VvSUC12.