Kumar Dilip, Kirti Pulugurtha Bharadwaja
Department of Plant Sciences, School of Life Science, University of Hyderabad, Hyderabad, India.
PLoS One. 2015 Feb 3;10(2):e0117559. doi: 10.1371/journal.pone.0117559. eCollection 2015.
Late leaf spot is a serious disease of peanut caused by the imperfect fungus, Phaeoisariopsis personata. Wild diploid species, Arachis diogoi. is reported to be highly resistant to this disease and asymptomatic. The objective of this study is to investigate the molecular responses of the wild peanut challenged with the late leaf spot pathogen using cDNA-AFLP and 2D proteomic study. A total of 233 reliable, differentially expressed genes were identified in Arachis diogoi. About one third of the TDFs exhibit no significant similarity with the known sequences in the data bases. Expressed sequence tag data showed that the characterized genes are involved in conferring resistance in the wild peanut to the pathogen challenge. Several genes for proteins involved in cell wall strengthening, hypersensitive cell death and resistance related proteins have been identified. Genes identified for other proteins appear to function in metabolism, signal transduction and defence. Nineteen TDFs based on the homology analysis of genes associated with defence, signal transduction and metabolism were further validated by quantitative real time PCR (qRT-PCR) analyses in resistant wild species in comparison with a susceptible peanut genotype in time course experiments. The proteins corresponding to six TDFs were differentially expressed at protein level also. Differentially expressed TDFs and proteins in wild peanut indicate its defence mechanism upon pathogen challenge and provide initial breakthrough of genes possibly involved in recognition events and early signalling responses to combat the pathogen through subsequent development of resistivity. This is the first attempt to elucidate the molecular basis of the response of the resistant genotype to the late leaf spot pathogen, and its defence mechanism.
花生晚斑病是由半知菌亚门真菌花生球腔菌引起的一种严重病害。据报道,野生二倍体物种Arachis diogoi对该病具有高度抗性且无症状。本研究的目的是利用cDNA-AFLP和二维蛋白质组学研究,调查受花生晚斑病病原菌侵染的野生花生的分子反应。在Arachis diogoi中总共鉴定出233个可靠的差异表达基因。约三分之一的转录衍生片段(TDFs)与数据库中的已知序列没有显著相似性。表达序列标签数据表明,所鉴定的基因参与赋予野生花生对病原菌侵染的抗性。已鉴定出几个与细胞壁强化、过敏细胞死亡和抗性相关蛋白有关的基因。鉴定出的其他蛋白相关基因似乎在代谢、信号转导和防御中发挥作用。在时间进程实验中,通过定量实时PCR(qRT-PCR)分析,对与防御、信号转导和代谢相关的基因进行同源性分析,在抗性野生种中进一步验证了19个TDFs,并与一个感病花生基因型进行比较。与六个TDFs相对应的蛋白质在蛋白质水平上也有差异表达。野生花生中差异表达的TDFs和蛋白质表明其在病原菌侵染时的防御机制,并为可能参与识别事件和早期信号反应以通过后续抗性发展对抗病原菌的基因提供了初步突破。这是首次尝试阐明抗性基因型对花生晚斑病病原菌反应的分子基础及其防御机制。