Molecular Cloning And Functional Analysis Of VpR82H Gene In Chinese Wild Vitis Pseudoreticulata

The grapevine is the fourth sequencing flowering plants after Arabidopsis,rice, and poplar. Thisprovides a good platform and resources for further accelerate resistance gene research.As one of the world’smost important fruit crop, the most large-scale cultivation of the Vitis vinifera varieties are generallysusceptible to Grape powdery mildew (Erysiphe necator)，which is a commolon fungal diseasethat widelydistributed in the world’s major grape-producing areas, it’s a serious threat to the grape production andprocessing. China， as one of the the origin and the most important areas of grapes, with a rich variety ofwild grape germplasm repository. and there are strong adaptability and resistance among Chinese wildgrapes.In this study, homology cloning technique was used to clone a broad-spectrum resistance geneRPW8.2Arabidopsis homology gene VpR82H from Chinese wild Vitis pseudoreticulata accession Baihe-35-1， Through sequence analysis and sequence alignment with European grapes after select from thesequences VpR82H in expression patterns in different tissues were analyzed to meet the requirements afteraccurately sequenced by semi-quantitative RT-PCR technique, VpR82H genes were expressed in varioustissues of different grapes, Real-time quantitative RT-PCR was found VpR82H gene was induced bybiological and exogenous signaling molecules. Grape powdery mildew and signaling molecules sprayingexogenous ABA, SA, MEJA are able to induce gene expression,0℃low temperature treatment mediatedthe same impact in VpR82H expression level; plant expression vectors with different promoters werebuilded, the VpR82H were transgenic into Arabidopsis by Agrobacterium-mediated gene methods ofFloral dipping.By using a fluorescent microscope screened T2gener ation of transgenic Arabidopsisseedlings was observed after inoculated powdery mildew,the expression of green fluorescent protein wasfouned.Same transient transformation after the benthamiana The GFP was also observed aroundpowdery mildew mycelium, indicating that the VpR82H gene expression can indeed be induced bypowdery mildew, and possiblly participated in the powdery mildew resistance response process.