Study On Proteomics Of Powdery Mildew Resistant And Susceptible Grape's Leaves

Powdery mildew is one of the fungi disease which could harm grapes seriously. Taking advantage of the genetic resources of the disease resistance is one of important methods to solve the problem. Protein is the essence of life action, different disease resistance of grape is determined by specific proteomics, the difference of proteomics can reflect the essence of the disease resistance of plant more directly than the difference of genomics. So, using the research methods of proteomics to compare and investigate the difference of proteomics of grape with different disease resistance, finding out disease-susceptible and disease-resistant protein, obtaining the genes were directly involved in the disease-resistance of powdery mildew, have very important theoretic significance and practical value in solving the problem of the disease in the production of grape.Disease-susceptible grape Carignane(the male parent) and disease-resistant grape Baihe-35-1(the female parent)'s F1, disease-resistant plant 6-12-4 and disease-susceptible plant 6-12-2 were the material of this research, Their leaves were inoculated by grape powdery mildew artificially and then picked 0 day, 2 days nd 4 day's aftern inoculation, their differences of proteomics of the grape's leaves with different disease-resistance were analysed.The primary results are showed as fo11ows:(1) The method system of 2-DE was established initially. The results of 2-dimensional electrophoresis from different extraction methods, different loading capictity of total sample protein, different procedures of isoelectric focusing, different size of immobilized pH gradients strips and with different SDS-PAGE gel concentration were compared. The results showed that protein loading volume of 300μg ,isoelectric focusing procedure B , and 12% SDS-PAGE gel were the better choice for 7 cm immobilized pH gradients (IPG) strip with the linear range of pH 4～7. Protein loading volume of 1050μg is suitable for 17 cm immobilized pH gradients (IPG) strip with the linear range of pH 4～7.(2) The differences of proteomics of the six kind of materials handled differently were compared. 26 protein spots with or without expression of difference were found,10 of them were recycled and purified, they were identified by MS, 7 of them were positive(1305,4708, 4709,5411,5625,6915,7121), the others(1222,1420,7615) were negative.(3) Spot 1305 and 6915 were hypothetical protein, spot 4708 and 4709 were unnamed protein product. Spot 5411 was oxygen evolving complex 33 kDa photosystem II protein; spot 5625 was ribulose-1,5-bis-phosphate carboxylase/oxygenase activase 1, Rubisco. They were somehow associated with the disease-resistant and disease-susceptible characteristics of grape.