The Research Of Drought Proteome Of Wheat Leaves Under Drought Stress

With the scarcity of freshwater resources and increased drought year after year, the threat of food production has been growing and the research of drought resistance is highly valued at home and abroad. Therefore, it laid theoretical foundation for the breeding of wheat varieties with drought resistance and drought resistance mechanism of wheat. Some scholars have been studied on expression of D-response of leaves of wheat seedling, results showing that the D-response protein related to the degree of drought resistance. Drought resistance of wheat is not single but the response of a variety of protein interaction or interaction with the environment. In order to find more proteins associated with drought resistance, this experiment will use two-dimensional electrophoresis analysis proteomic of wheat leaves after drought stress, establishing the theoretical foundation for the mechanism of drought resistance. Fully and rationally utilize water resources, breeding wheat varieties with more drought resistance. The main are as follows:(1) A 2-DE optimization technology system for wheat leaf holoprotein was developedWith different drought resistance winter wheat varieties Xinong 2208 and Xinong 189 for the materials, compared the effects of two different protein extraction method (TCA/acetone precipitation and sodium phosphate buffer grinding method) on two-dimensional electrophoresis technology after PEG stress during seeding stage, and the PH range of IPG gel strip, the loading quantity of samples and the gel concentrations were optimized. The result showed that the expression of RuBisCO enzyme reduced with TCA / acetone extraction total protein of leaf and better separation of molecular weight proteins and low abundance proteins. The proteins were separated obviously with pH 4~7 and 17 cm IPG line strip in the isoelectric point and molecular weight. A better 2-DE map was adopted by two-dimensional electrophoresis technology with 150 g/ gel loading quantity and 12% concentrations of SDS-PAGE gel. Analysised the difference of the protein map, twenty-eight protein spots in Xinong 189 and Xinong 2208 were present difference , In which a low credibility; including twenty-two down-regulated or up-regulated; including five novel induced expression.(2) analysis and identification difference proteinsIn order to reveal the molecular genetic mechanisms of wheat drought stress in protein levels, extracted with TCA / acetone the whole protein samples of leaves, separated and silvered staining it by solid-phase IEF / SDS-PAGE two-dimensional gel electrophoresis, has been reproducible two-dimensional electrophoresis map. Identified about 290 clearer protein spots with the PDQuest-DE software,28 differentially expressed proteins identified by MALDI-TOF, among which 22 proteins with apparent specificity respectively, are: 1,5 - ribulose bisphosphate carboxylase/oxygenase, acetyl coenzyme A carboxylase, chalcone synthase, 5 unknown proteins and defense mechanisms protein and so on.Analysising protein clustering of proteins amino acid sequence by mass spectrometric identification with bioinformatics and MEGA4.0 software, mostly unknown proteins are related to Calvin cycle and glycolysis. The proteins are directly involved in hormonal regulation, photosynthesis, protein transport, protein interactions, resilience defense of wheat leaves. Drought stress induced potential gene expression in wheat leaves, and the genes control drought resistance protein function and physiological metabolic process.