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

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.