小麦应答条锈病菌冬孢子产生的蛋白质组学分析  

Proteomic Analysis of Wheat(Triticum aestivum) Induced by Puccinia striiformis f.sp.tritici at Telial Stage

作　　者：万翠萍[1] 李辉[1] 王付平[1] 康振生[1] 韩青梅[1] 

机构地区：[1]西北农林科技大学植物保护学院/旱区作物逆境生物学国家重点实验室,杨凌712100

出　　处：《农业生物技术学报》2016年第6期815-823,共9页Journal of Agricultural Biotechnology

基　　金：国家重点基础研究发展规划(973)项目(No.2013CB127700);高等学校学科创新引智计划(No.B07049)

摘　　要：由小麦条锈菌(Puccinia striiformis f.sp.tritici,Pst)引起的小麦条锈病是我国小麦(Triticum aestivum)最严重的流行病害之一。中国小麦条锈菌表现出高度的遗传多样性和毒性变异,现已研究表明小麦条锈菌经有性生殖可发生致病性变异并产生新的小种。冬孢子作为小麦条锈菌有性阶段的开始,是小麦条锈菌生活史中完成有性生殖过程必不可少的阶段。为分析小麦条锈菌产冬孢子时期寄主植物的叶片总蛋白表达变化,探究寄主植物如何响应冬孢子产生,本研究利用三氯乙酸(trichloroacetic acid,TCA)/酚抽提法提取产冬孢子时期和同时期未接种的小麦叶片总蛋白,经双向电泳技术分离后,利用PDQuest软件进行分析并选取显著上调表达的蛋白进行Orbitrap质谱鉴定。结果表明,在产冬孢子时期的寄主叶片总蛋白图谱中发现22个蛋白点(上调表达大于1.5倍且符合t检验),经功能注释发现这些差异蛋白参与糖代谢、抗逆境以及植物衰老相关代谢途径。之后利用q RT-PCR技术对蛋白水平的结果进行验证,结果发现这22个蛋白点中有16个差异蛋白的基因在转录水平上调表达(相对表达倍数大于1.5倍),证实了蛋白结果的可靠性。本研究通过双向电泳技术发现小麦条锈菌冬孢子的产生影响了寄主叶片参与糖代谢、抗逆境及植物衰老相关代谢过程蛋白的表达,并推测小麦条锈菌冬孢子的产生可能与寄主植物的衰老有关。本研究为进一步深入了解小麦条锈菌冬孢子阶段寄主植物与病原菌的互作机制提供了一定的理论基础,对于揭示冬孢子的形成机制和全面了解小麦条锈菌的有性阶段有重要意义。Wheat stripe rust or yellow rust, caused by an biotrophic fungus Puccinia striiformis f. sp. tritici （Pst）, is one of the important epidemic disease of wheat （Triticum aestivum） in China. Wheat stripe rust in China has shown high levels of genetic diversity and toxicity mutation. Research has shown that sexual reproduction of Pst can result in pathogenic mutation and produce new races. As the start of the sexual stage of wheat stripe rust, teliospore is an essential stage for completing Pst sexual reproduction. To analyse the proteomic changes of wheat leave infected by Pst at telial stage and investigate how the host plant respond to the sporification of teliospore. Proteins were extracted from leaves infected with stripe rust CYR32 and control group using trichloroacetic acid （TCA）/phenol extraction, and then separated by using two-dimensional electeophoresis （2-DE）. Using PDQuest software analysis, 22 up-regulated spots （relative expression more than 1.5 folds） were observed in Suwon11 leaves inoculated with stripe rust CYR32 at telial stage. The up-regulated protein spots were selected for Orbitrap analysis and database searching. All these proteins were successfully identified and were mainly involved in glucose metabolism, anti-stress and plant senescence related metabolic pathways. There were 13 proteins involved in glucose metabolism, among which triose phosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were key enzymes in the process of glycolysis, and isocitrate dehydrogenase participates in the Krebs cycle. Five proteins were involved in polysaccharide degradation and related metabolic, with the rest participating in ATP synthesis. There were 6 anti-stress related proteins, mainly involved in the removal of harmful reactive groups and the synthesis of osmolytes. The rest 3 proteins participate in plant senescence related metabolic pathways, among which both ATP-dependent Clp protease proteolytic subunit and cysteine protease precursor participate in the degradation of

关 键 词：小麦条锈菌 冬孢子 植物衰老 蛋白质组学 双向凝胶电泳 

分 类 号：S432.1[农业科学—植物病理学]