大豆应答玉米小斑病菌胁迫的蛋白质双向电泳分析

Proteomic Analysis of Glycine max Leaf in Response to Stress of Bipolaris maydis

为探究大豆应答非致病菌(Bipolaris maydis)胁迫的分子机制,将玉米小斑病菌接种于3周龄的大豆苗。取48 hpi的大豆叶片以TCA/丙酮法提取蛋白质,利用蛋白质双向电泳技术分离蛋白,串联质谱(MS+MS/MS)鉴定蛋白质再结合生物信息学技术进行蛋白质功能分析。结果显示:分离到大豆苗期叶片总蛋白点1 300±15个,其中,差异表达(2倍以上,P值<0.05)的蛋白质点18个,除2个蛋白点未成功测序,其余16个差异蛋白点经GO分析可知,差异表达的蛋白质功能主要涉及光合作用、胁迫应激和非寄主抗性响应,如核酮糖1,5-二磷酸羧化酶、病程相关蛋白PR-10和细胞骨架结构相关蛋白profilin-2。同时,由测序结果推测:在非致病菌(玉米小斑病菌)的胁迫下,大豆通过降低光合作用相关蛋白和能量代谢相关蛋白的表达,来增强防御相关蛋白和非寄主抗性相关蛋白的表达。

To investigate the response of the Glycine max to non-adapted fungi stress, 3-week-old soybean seed- lings were subjected to Bipolaris maydis stress. Leaf tissues were harvested 48 h post inoculated and TCA-acetone precipitation method was used for protein extraction. A tandem combination of two-dimensional electrophoresis, mass spectrometry （MS＋MS/MS）, and bioinformatics tools were used to identify protein that contribute to nonhost resistance in Glycine max. Results showed almost 1300 protein spots were separated with 2-DE gels of leaf of Glycine max. Eighteen differentially proteins （Iratiol〉2, p〈0.05） were identified in inoculated compared to controlling plants, including proteins involved in photosynthesis, response to stress, and non-host resistant response such as RuBisCO, pathogen related protein PR-10, and cytoskeleton structure related protein profilin-2. Taken together, when Glycine max was subjected to the non-adapted pathogen Bipolaris maydis, it expressed non-host resistance （NHR） 48 hpi by up or down-regulating the defense related proteins. The results also suggested that the decreased expression of protein involved in photosynthesis and energy could be beneficial to the increased expression of protein involved in defense response and non-host resistant response.