Proteomics-based Analysis Of Low Abundance Proteins In Leaves Of Wheat NILs Taichung 29*6/Yr5

Wheat stripe rust is one of the most important wheat leaf diseases caused by fungus Puccinia striiformis f.sp. tritici, which is responsible for major damage in wheat and result in both yield losses and downgrading in quality worldwide. Use of resistant cultivars is the most economical, effective and safe way to control wheat stripe rust. Because of simplification of resistant cultivars and development of the new physiologic races, the resistance genes frequently became susceptible. Therefore, it is greatly important theoretically and practically to understand the disease resistant mechanism for improving disease resistance in wheat. In the leaves of plants, the low-abundant proteins such as regulatory and signaling factors, and other rare proteins are related to the resistance and physiological mechanisms of plants. Due to the developing methods and demand of research, the analysis of low abundance proteins has been an effective way to reveal the physiological mechanism.In this study, we used the susceptible recurrent parent Taichung 29 as a control, to compare the differences of expressed low abundance proteins of leaves of the wheat stripe rust resistance isogenic line Taichung29*6/Yr5 supplied by Institute of plant protection of Chinese Academy of Agricultural Sciences. Through analyses of the changes of low abundance proteins in Taichung29*6/Yr5 after inoculated, we try to find specific-expressed resistance-related proteins.In this study, The proteins were extracted from leaves by buffer solution method. The sample was separated through an ion-exchange column. After SDS-PAGE analysis, fractions eluted between 0.10-0.25 M NaCl were detected to contain most of low abundance proteins. The result of 2-DE showed that, the separation system based on AEX was efficient and could be applied for analysis of the low abundance proteome in wheat leaf.The total proteins of Taichung29 and Taichung29*6/Yr5 leaves were extracted after inoculated by the epidemic race CYR32. The high abundance proteins were removed by anion exchange chromatography. SDS-PAGE and 2-DE were used to compare the differences of low abundance proteins. As a result,5 and 11 protein spots were found differentially expressed in Taichung 29 and Taichung29*6/Yr5 after inoculated respectively. There were 20 different spots detected in the comparison between two inoculated materials. 25 peptide mass fingerprints (PMFs) were obtained of the specific-expressed proteins after identified by MALDI-TOF-MS analysis.12 proteins were identified by against NCBI databases. The result shows that, the specific-expressed proteins were related to glycometabolism, glycolipid metabolism and other hardiness physiology of plants. Some of them are beta-glucosidase, tetratricopeptide repeat-containing protein, lipoxygenase, ascorbate peroxidase,3-phosphoglycerate kinase. These functions are mainly involved in plant defense responses. They not only participated in repair trauma caused by stripe rust fungus, but also inhibit the growth of stripe rust fungus. Base on the experiment results, the obtained data were provide theoretical principle and gene resources for molecular breeding of wheat resistance to puccinia striiformis, and helpful to the resistant mechanisms research for similar plant samples.