大豆疫霉PsMPK1沉默突变体的基因表达谱分析  

Gene expression profiling of a Ps MPK1- silenced mutant of Phytophthora sojae

作　　者：叶文武[1] 张萌[1] 曹明娜[1] 翟春花[1] 李爱宁[1] 王源超[1] 

机构地区：[1]南京农业大学植物保护学院,江苏南京210095

出　　处：《南京农业大学学报》2016年第3期386-393,共8页Journal of Nanjing Agricultural University

基　　金：中央高校基本科研业务费专项资金项目(KJQN201507);国家自然科学基金项目(31401688);江苏省自然科学基金项目(BK20140698);国家国际科技合作专项资助(2013DFG32030)

摘　　要：[目的]促有丝分裂原蛋白激酶(MAPK)是真核生物细胞信号转导途径中的关键蛋白激酶,对植物病原菌的生长发育与侵染致病具有重要作用。我们前期发现1个MAPK基因Ps MPK1参与调控大豆疫霉的细胞壁完整性、菌丝生长、游动孢子形成,以及对大豆的致病性等生物学性状。本文旨在进一步探究Ps MPK1如何调控大豆疫霉的上述性状。[方法]利用数字基因表达谱技术对大豆疫霉Ps MPK1沉默突变体在游动孢子囊阶段的基因表达情况进行分析。[结果]样品间基因表达水平的整体相关性分析和实时定量PCR结果表明:数据具有较高的可靠性。与野生型菌株相比,Ps MPK1沉默突变体中分别有1 451个显著下调和981个显著上调的基因,占所有被检测基因的23%。与之相比,被检测的细胞壁相关基因和分泌蛋白编码基因中,显著差异表达的基因比例更高(分别占37%和28%),且大部分基因为下调表达(分别占87%和78%)。显著下调表达的分泌蛋白编码基因中,37个基因可能与侵染过程相关,主要编码Rx LR、NLP和CRN等家族的效应分子,CBEL蛋白,富含半胱氨酸蛋白,氧化还原相关蛋白以及水解酶等。[结论]首次通过转录组学方法揭示了特定的功能基因Ps MPK1沉默后大豆疫霉基因组的转录变化,鉴定了可能与Ps MPK1信号途径相关的基因和基因家族,为进一步研究大豆疫霉中MAPK的调控网络奠定了基础。[Objectives]Mitogen-activated protein kinase（ MAPK） is a class of important protein kinases in signal transduction pathways of eukaryotic cells,and involved in growth,development,and pathogenesis of plant pathogens. We previously identified a MAPK gene called Ps MPK1,which is required for cell wall integrity,hyphal growth,zoosporogenesis,and pathogenicity in Phytophthora sojae. The purpose of this study is to further learn how Ps MPK1 regulates the abovementioned processes. [Methods]We profiled gene expression changes using digital gene expression（ DGE） profiling method for a Ps MPK1-silenced transformant at zoosporangia stage. [Results]Both correlation analysis of whole-library gene expression patterns and qRT-PCR validation results suggested a robustness of the DGE data.In comparison with the wild type isolate of P. sojae,23% of the all detected genes were differentially expressed in the Ps MPK1-silenced transformant,including 1 451 up-regulated and 981 down-regulated genes. 37% of cell wall-associated genes and 28% of secreted protein encoding genes exhibited differential expression,and 87% and 78% of them were down-regulated,respectively. Of the downregulated secreted protein encoding genes,37 were putative pathogenicity-related genes. These genes were predicted to encode Rx LR,NLP,and CRN effectors,CBEL proteins,cysteine-rich proteins,redox-related proteins,and hydrolases. [Conclusions]This is the first global transcriptome analysis for a P. sojae mutant defected in Ps MPK1. We identified candidate genes and gene families related to Ps MPK1 signal transduction pathways,which will provide basis for further study of MAPK-regulated network.

关 键 词：大豆疫霉 促有丝分裂原蛋白激酶 细胞壁 致病性 效应分子 转录 

分 类 号：Q945.8[农业科学—植物病理学]