大豆花叶病毒诱导的野生大豆抑制差减文库构建及初步分析

Construction and Primary Analysis of a SSH Library of Wild Soybean Induced by SMV

以野生大豆Bian0526为试验材料,通过抑制性差减杂交技术(SSH)构建了大豆花叶病毒(SMV)诱导的cDNA差减文库。在文库中共筛选到15 343个阳性克隆,PCR鉴定插入片段大部分集中在500～800 bp之间。利用BLAST在GenBank文库中进行序列相似性比对,获得有功能的EST 200个。对有功能注释的同源分析发现,其功能涉及植物的细胞自身保护和系统获得性抗性等方面,说明这些基因很可能参与到野生大豆抗大豆花叶病毒反应中。

To isolate the genes related to wild soybean resistance in Soybean Mosaic Virus（SMV）,Suppression Subtractive Hybridization（SSH）was utilized to construct the different expression cDNA library using wild soybean variety Bian0526 inoculated with race 3 of SMV.From this library,a total of 15 343 positive clones were screened,and the length of most of the insert fragments was focused on 500-800 bp.ESTs similarity analysis of these unigenes was performed with the Blastx and Blastn against the non-redundance database in GenBank.The results showed that 200 functional ESTs had been gotten,and they were related to cell self-protection and system acquired resistance,etc.This indicated that these genes might be involved in resistance to SMV of wild soybean.