Expression Of Candidate Resistance Gene Induced By Powdery Mildew And VpWDR Gene Cloning And Localization

Chinese wild resistant grape species are important resist germ plasm resource. It'ssignificant to separate and identity resistant genes for disease-resistant mechanism andcultivating disease-resistant varieties. This research used Chinese wild Vitis pseudoreticulatacv. baihe35-1leaves to study the candidate resistant genes expression induced by powderymildew through the real-time quantitative PCR, clone the resistance-related gene VpWDR andstudy the function of the gene. The main results are as follow：1. After induced by powdery mildew, the expression of17candidate gene was studied byreal-time quantitative PCR. The result showed that the17candidate genes expression presentdouble peak phenomenon, single peak phenomenon, increase by degrees and decrease bydegrees.13genes were up regulated,4genes were down regulated. The largest expressionwas2.8times accronding to the control in up regulated expression genes; the least expressionwas0.13times accronding to the control in down regulated expression genes.2. A full length gene was cloned using RACE technology, the gene was named VpWDR.It contained1577bp, the longest open reading frame was1377bp in length encoding458Amino acids, deduced pI was5.07, and molecular weight was51.7KD. The gene belongs toWD40gene family. The plants fusion express carrier VpWDR/pBI221-GFP was transfer intoepiderm of Allium by gene gun.The localization of the gene showed that the translated proteinlocated at cell nucleus, cytoplasm and cell membrane.