Proteomic Analysis Of Wheat Stripe Rust Resistance Near Isogenic Line Taichung29~*6/Yr5

Wheat sripe rust, caused by the fungus Puccinia striiformis West. f.sp. tritici Eriks, is one of the most important wheat diseases worldwide. It can lead to nearly 40 percent of yield loss in some fields in epidemic years. Nowadays, the wide application of resistance cultivars is the major strategy for wheat stripe rust control. Use of resistant cultivars has become the major means of controlling stripe rust and resistant cultivars utilization is the most effective, economical and safe way. Because of development of the new physiologic races, wheat stripe rust resistance genes frequently became susceptible. Therefore, It's very important for preventing and controlling wheat sripe rust that found and clones the stripe rust resistance gene in the short time.Recently, wheat stripe rust has been studied on many aspects, such as morphology, cytology, physiological and biochemical prodesses, but the molecular mechanism of wheat stripe rust resistance was little understood. In recent years, proteomics, as an important new research method of functional genomics, has been wide applicated.In this study, we studied wheat stripe rust resistance to the pathogen Puccinia striiformis on molecular level with proteomic method aiming at finding the interaction mechanisms between wheat host and its pathogen Puccinia striiformis. The results will be beneficial to wheat resistance breeding.We used the wheat stripe rust resistance gene Yr5 near isogenic line(NIL) Taichung29*6/Yr5, and its susceptible recurrent parent Taichung 29 as a control supplied by Institute of plant protection of Chinese Academy of Agricultural Sciences.In order to compare the differences of expressed wheat proteins after inoculation with pathogen, we used the two-dimensional electrophoresitic technology to analyze the differences of expressed total proteins of Taichung29*6/Yr5 and its control vatiety Taichung29 inoculated with the race CY32 of Puccinia striiformis f.sp. tritici 2 ,5 and 7 days later, sepecially the changes of expressed proteins after after 5 days of inoculation. After 2 days of inoculation, we found 13 specific-expressed proteins in Taichung29*6/Yr5; After 7 days of inoculation, we found 28 specific-expressed ones; After 5 days of inoculation, we found 45 specific-expressed proteins in the four groups of materials. These specific-expressed prpteins were of significant difference of protein spot and the percentage of volume change more than 1.5-2 times according to the T-test (p <0.05).Through matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS ; MALDI-TOF-TOF MS/MS), we got these specific-expressed protein mass fingerprintings (PMFs) and their amino acid sequences. By searching the related plant protein database using the MASCOT search engine, we found these proteins were related to disease resistance, metabolism, photosynthetic electron transport etc. Some of them are pathogenisis-related and defense proteins of SAR, such as pathogenisis-related protein 1.2 (PR-1.2), beta-1,3-glucanase, glutathione S-transferase, E3 ubiquitin ligase, ferredoxin-NADP(H) oxidoreductase etc.Based on the experimental results, we tentatively concluded that 2-DE could be an effective method in future wheat leaf proteomics analysis, and application of 2-DE can acquire more specific-expressed proteins; proteomic analysis of NIL Taichung29*6/Yr5 will be beneficial to elucidating wheat resistance mechanisms to stripe rust and providing theoretical principles and gene resources for wheat molecular resistance breeding to Puccinia striiformis.