青枯菌诱导的花生基因表达谱SSH分析  

Analysis of Gene Expression Profiles in Response to Ralstonia solanacearum Using SSH in Peanut

作　　者：吕建伟[1,2] 姜慧芳[1] 黄家权[1] 彭文舫[1] 陈玉宁[1] 李正强 

机构地区：[1]中国农业科学院油料作物研究所/农业部油料作物生物学重点实验室,武汉430062 [2]贵州省农作物品种资源研究所,贵阳550006

出　　处：《西北植物学报》2011年第8期1517-1523,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家公益性科研院所专项资金课题(2007-05)

摘　　要：以抗青枯病花生种质‘J4’和‘中花6号’、感青枯病花生品种‘中花12号’为材料,用强产青枯菌毒菌株(Ralstonia solanacearum)对其根系分别接种,采用抑制差减杂交(SSH)技术检测花生根系应答侵染的基因表达谱变化,并对文库中差异基因进行Real-time PCR分析。结果表明:经菌液PCR检测对挑选出的1 036阳性克隆片段进行测序及片段整合分析,获得162条花生基因,有功能注释的基因58条,其中44条基因参与了细胞结构(6%)、信号转导(12%)、抗病防御(5%)、转录调控(12%)等生理过程。用Real-time PCR技术对7个基因在‘中花6号’和‘中花12号’中的表达模式分析结果表明,6个基因在青枯菌侵染早期在抗病材料‘中花6号’中呈上调表达,可能与青枯病抗性直接相关。Bacterial wilt caused by Ralstonia solanacearum is one of the most bacterial diseases in many countries including China.Gene expression changes associated with resistance to the infection by R.solanacearum were detected in some peanut roots with various resistances including genotypes ＇J4＇ and ＇Zhonghua 6＇ both with resistance and genotype ＇Zhonghua 12＇ with susceptible.Based on the results from 1 036 clones randomly selected from the cDNA library were sequenced and then clustered and assembled.162 gene fragments were obtained including 58 fragments with function annotation,of which 44 involved in the progress of cell structure,signal transduction,defense response and gene transcription accounted for 6%,12%,5%,12%,respectively.Further,the expression profiles of 7 gene fragments selected from the 44 functional genes were detected in the resistant genotype ＇Zhonghua 6＇ and susceptible genotype ＇Zhonghua 12＇ by using Real-time PCR.The results showed that 6 genes of them were upregulated in the early stage after R.solanacearum infection which might be contributed to peanut resistance to R.solanacearum.The result from the present study provided a foundation for cloning the resistance genes and investigating the molecular mechanism of peanut resistance to R.solanacearum.

关 键 词：花生 青枯菌 SSH文库 RT-PCR 

分 类 号：Q789[生物学—分子生物学]