Proteomic Analysis Of Leaf Intercellular Fluid From Stripe Rust Resistance Wheat

As a main crop in China, Wheat(Triticum aestivum) is planted in area of about 24 million hectares, the yield of which is about 20% of national grain output. Unfortunately, the wheat stripe rust(Puccinia striiformis f. sp.tritici) which has been a major disease of wheat has been threatening with China's wheat production and food security. It is important theoretical and practical significance to study wheat stripe rust and the interaction between wheat and stripe rust, so that it can play a key role in application of wheat disease-resistant cultivar and the prevention of wheat stripe rust.In this paper, the wheat near-isogenic lines which included gene Yr5 conferring high resistance to stripe rust were chose as experimental material, the main research contents include proteomics Analysis of intercellular washing fluids (IWF) in wheat leaf of Taichung29 and Taichung29*6/Yr5 which was infected by stripe rust CYR32 and the influence of Stripe rust fungus spores which germinate in IWF. The results are as follows:Firstly, we established a system for Separation and analysis of IWF, and comparative analysis of IWF protein and total protein in wheat leaf. When separated using 2-DE, in IWF protein images, there were 1042±47 particular spots when compared with total protein images. The MALDI-TOF-MS analysis of 100 spots that were randomly chosen from IWF protein 2-DE gels. The identified proteins include some pathogen-related proteins such asβ-1,3-glucanase, glucan endo-1,3-β-D-glucosidase, chitinase, peroxidase and so on.Secondly, we further researched on proteomics Analysis of IWF in wheat leaf of Taichung29 and Taichung29*6/Yr5 which was infected by stripe rust CYR32 after 36 hours, we found proteomics of intercellular fluids in wheat leaf had some obvious changes.23 extremely significant differential protein spots were detected in Taichung29*6/Yr5 after infected, and 31 extremely significant differential protein spots in Taichung29 after infected. In two types differential protein spots,5 differential protein spots were common.17 extremely significant differential protein spots were detected in Taichung29*6/Yr5 and Taichung29 after infected, and 13 extremely significant differential protein spots before infected. In two types differential protein spots, the common spots couldn't be found. By analysis, we infer that these proteins are related to infection of wheat stripe rust and function of wheat resistance to stripe rust, but there were not high identification rate by MALDI-TOF/TOF.Preliminary analysis showed that intercellular fluids in Taichung29 and aichung29*6/Yr5 had an obvious influence on germination of Stripe rust fungus spores. We found germination rate of wheat would recovery normal when the concentration of intercellular fluids of Taichung29 and aichung29*6/Yr5 were diluted from 0.2mg/ml to 0.02mg/ml. In addition, intercellular fluids had a large effect on germ tube of urediospores.