利用cDNA-AFLP技术分析小麦成株抗条锈性差异基因表达特征  

cDNA-AFLP Analysis Reveals Differential Gene Expression in Wheat Adult-Plant Resistance to Stripe Rust

作　　者：张岗[1,2,3] 董艳玲[1] 夏宁[1] 张毅[1,4] 王晓杰[1] 屈志鹏[1] 李依民[1] 黄丽丽[1] 康振生[1,5] 

机构地区：[1]西北农林科技大学植物保护学院,陕西杨凌712100 [2]陕西中医学院药学院 [3]中国医学科学院北京协和医学院药用植物研究所 [4]陕西省西安市农业技术推广中心植保站 [5]西北农林科技大学/陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《作物学报》2010年第3期401-409,共9页Acta Agronomica Sinica

基　　金：国家自然科学基金重点项目(30930064);现代农业产业技术体系建设专项资金;高等学校学科创新引智计划项目(B07049);国家“十一五”科技支撑计划项目(2006BAD08A05)资助

摘　　要：采用cDNA-AFLP技术,对成株抗条锈小麦品种兴资9104在成株期受条锈菌生理小种CY32侵染后5d内9个时间点的基因表达谱进行了分析。共筛选64对引物,产生32320个转录本(TDF);用37对引物检测到2201个(6.81%)差异TDF,其中926个TDF诱导表达,1275个下调表达。经大规模克隆、测序分析,最终获得330个差异TDF,聚类分析得到259个EST(unigenes),命名为aTaPST1至aTaPST259(GenBank登录号为FL645754～FL646011和FL646262)。经Blastx比对和功能分类分析,其中96条EST(37.07%)未找到同源性匹配,68条(26.25%)与未知功能蛋白同源性较高;其余95条ESTs主要涉及能量(11.20%)、基础代谢(4.63%)、转录调控(3.86%)、抗病与防御(3.86%)、蛋白质运输和储存(3.09%)、蛋白质合成和细胞生长(各2.32%)以及信号转导(1.54%)等。选取抗病与防御、转录调控及信号转导类等相关的6个差异基因,qRT-PCR分析结果显示其表达模式符合cDNA-AFLP表达谱。小麦成株抗条锈性分子机制涉及植物多方面生理生化反应,包括抗病与防御、转录调控、蛋白质代谢、信号转导、以及非生物胁迫等多种途径相关基因的协同控制。Wheat （Triticum aestivum L.） stripe rust is one of the most devastating diseases of wheat throughout the world. Adult plant resistance （APR） to stripe rust in wheat conferring durable resistance, thus, plays a pivotal role in the control of the disease. In the present study, to elucidate the molecular mechanism of wheat APR to stripe rust, we conducted extensive transcription profiling of adult-plant wheat cultivar Xingzi 9104 infected by Puccinia striiformis Westend f. sp. tritici Erikss. pathotype CY32 using cDNA-AFLP technique. We analyzed transcription profiling of the incompatible reaction across nine sampling time points within five days after inoculation. Of the total 32 320 transcript derived fragments （TDFs） obtained using cDNA-AFLP with 64 primer pairs, 2 201 （6.81%） displayed altered expression patterns after inoculation, of which 926 showed up-regulated and 1 275 down-regulated. Three hundred and thirty differentially expressed TDFs produced reliable sequences after cloning and sequencing, of which 259 expressed sequence tags （ESTs） of unigenes were obtained after assembling, designated from aTaPST1 to aTaPST259, deposited in GenBank with accessions numbers from FL645754 to FL646011 and FL646262. Blastx analyses and functional annotations were then performed and the results revealed that the 95 ESTs had predicted gene products mainly impli-cated in energy （11.20%）, metabolism （4.63%）, transcription （3.86%）, disease/defense （3.86%）, protein destination and storage （3.09%）, protein synthesis and cell growth （each accounted for 2.32%）, and signal transduction （1.54% of the sequenced total 259 ESTs）. Six differential genes related to disease/defense, transcription, and signal transduction were chosen for further qRT-PCR expression patterns, which confirmed the cDNA-AFLP profiles. Our results indicated that wheat APR to stripe rust involved in multifaceted biochemical and physiological reactions, including concerted regulation of the genes involved in differe

关 键 词：小麦 条锈菌 成株抗病性 基因表达 CDNA-AFLP QRT-PCR 

分 类 号：S512.1[农业科学—作物学]