大豆疫霉根腐病菌诱导下SSH文库构建及初步分析  

Construction and Analysis of A SSH Library of Soybean Upon Infection with Phytophthora sojae

作　　者：张淑珍[1] 徐鹏飞[1] 吴俊江 李文滨[1] 陈维元[3] 陈晨[1] 于安亮[1] 王金生[1] 靳立梅[1] 李岑[1] 

机构地区：[1]东北农业大学大豆研究所,大豆生物学教育部重点实验室,黑龙江哈尔滨150030 [2]黑龙省农业科学院大豆研究所,黑龙江哈尔滨150086 [3]黑龙江省农业科学院绥化农科所,黑龙江绥化152052

出　　处：《大豆科学》2008年第3期543-545,共3页Soybean Science

基　　金：国家自然科学基金资助项目(30400285,30671317);黑龙江省教育厅新世纪人才基金资助项目(NCET-06-007);中国博士后基金资助项目(20060400835);黑龙江省高校骨干教师资助项目(1151G007);黑龙江省农科院博士后基金资助项目(LRB06-010);农业部寒地重点开放实验室开放课题;博士后落户黑龙江启动基金资助项目(LBH-Q05032);黑龙江省青年科学基金资助项目(QC06C012)

摘　　要：大豆疫霉根腐病是危害大豆生产的毁灭性病害之一，研究利用抑制性消减杂交技术（Suppression Subtractive Hybridization，SSH）成功构建了疫霉菌诱导的大豆抗病品种绥农10差异表达的cDNA消减文库，从文库中共筛选到2067个阳性克隆，PCR鉴定插入片段大部分集中在100—800bp之间，利用BLAST在GenBank数据库进行序列相似性比对，获得有功能的EST375个，分析表明这些EST功能涉及大豆的抑制病原菌生长、细胞自身保护、信号传导、系统获得抗性、蛋白质合成、呼吸作用等。Phytophthora root and stem rot of soybean（ Phytophthora sojae） is one of destructive diseases endangering soybean production, but there have been few reports on gene expression after inoculated with pathogen. In this study , SSH cDNA library of resistance soybean variety Suinong 10 inoculated with race 1 of P. sojae was constructed successfully, and 2 067 positive clones were screened. The length of most of the insert fragments was focused on 100- 800 bp. 375 functional ESTs have been gotten, and they dealt with the effect of inhibition pathogen growth, cell self-protection, signal transduction, system acquired resistance,protein synthesis and respiration,etc, after compared with sequences in unigene database of GeneBank with BLASTn and BLASTx algorithm.

关 键 词：大豆疫霉根腐病 抑制消减杂交技术(SSH) CDNA文库 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]