Khan Najeeb Ullah, Liu Mengjie, Yang Xiufen, Qiu Dewen
The State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agriculture Sciences (CAAS), Beijing, 100081, China.
Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, Pakistan.
PLoS One. 2016 Jun 27;11(6):e0158112. doi: 10.1371/journal.pone.0158112. eCollection 2016.
MoHrip2 Magnaporthe oryzae hypersensitive protein 2 is an elicitor protein of rice blast fungus M. oryzae. Rice seedlings treated with MoHrip2 have shown an induced resistance to rice blast. To elucidate the mechanism underlying this MoHrip2 elicitation in rice, we used differential-display 2-D gel electrophoresis and qRT-PCR to assess the differential expression among the total proteins extracted from rice leaves at 24 h after treatment with MoHrip2 and buffer as a control. Among ~1000 protein spots detected on each gel, 10 proteins were newly induced, 4 were up-regulated, and 3 were down-regulated in MoHrip2-treated samples compared with the buffer control. Seventeen differentially expressed proteins were detected using MS/MS analysis and categorized into six groups according to their putative function: defense-related transcriptional factors, signal transduction-related proteins, reactive oxygen species (ROS) production, programmed cell death (PCD), defense-related proteins, and photosynthesis and energy-related proteins. The qPCR results (relative expression level of genes) further supported the differential expression of proteins in MoHrip2-treated rice leaves identified with 2D-gel, suggesting that MoHrip2 triggers an early defense response in rice leaves via stress-related pathways, and the results provide evidence for elicitor-induced resistance at the protein level.
稻瘟病菌过敏蛋白2（MoHrip2）是稻瘟病菌（Magnaporthe oryzae）的一种激发子蛋白。用MoHrip2处理的水稻幼苗对稻瘟病表现出诱导抗性。为了阐明水稻中这种MoHrip2激发作用的潜在机制，我们使用差异显示二维凝胶电泳和定量逆转录聚合酶链反应（qRT-PCR）来评估在用MoHrip2和作为对照的缓冲液处理24小时后从水稻叶片中提取的总蛋白之间的差异表达。在每块凝胶上检测到的约1000个蛋白点中，与缓冲液对照相比，在MoHrip2处理的样品中有10种蛋白被新诱导，4种上调，3种下调。使用串联质谱（MS/MS）分析检测到17种差异表达蛋白，并根据其假定功能分为六组：防御相关转录因子、信号转导相关蛋白、活性氧（ROS）产生、程序性细胞死亡（PCD）、防御相关蛋白以及光合作用和能量相关蛋白。定量聚合酶链反应（qPCR）结果（基因相对表达水平）进一步支持了二维凝胶鉴定的MoHrip2处理的水稻叶片中蛋白质的差异表达，表明MoHrip2通过与胁迫相关的途径触发水稻叶片的早期防御反应，并且这些结果为激发子诱导的抗性提供了蛋白质水平的证据。
