草业学报2017，Vol.26Issue(12)：152-159,8.DOI:10.11686/cyxb2017063
马铃薯病毒诱导应答基因抑制消减杂交文库构建及分析
Construction and analysis of a suppression subtractive hybridization library for the potato virus induced response gene
摘要
Abstract
Potato virus accumulation is one of the important reasons for the decline of yield and quality in potato production.In order to screen and clone the potato virus induced response gene,we constructed a suppression subtractive hybridization (SSH) library using cDNAs from potato virus disease carrying plant leaf as the tester,and those cDNAs from a virus-free seedling leaf as the driver.To verify the effect of library,98 randomly selected positive clones were identified by PCR from the library and sequenced,and 45 high quality non repeat sequences were obtained.By blast analysis in GenBank,we found that 14 of them were sequences belonging to the potato virus gene,22 of them had high homology with known genes,and 9 of them had no homologous reference gene.Two frequently found ESTs in the library were analyzed using quantitative real time PCR,and their expression was induced by potato virus.The results therefore showed that the SSH library was constructed successfully.This work has thus laid a foundation for further screening for potato virus related pathogenicity,and induction of corresponding defense response genes.This may lead to analysis of the molecular mechanisms of interaction between the potato plant and the virus,and use of biotechnology to cultivate virus resistant potato varieties.关键词
马铃薯/病毒/应答基因/抑制消减杂交Key words
potato/virus/response gene/suppression subtractive hybridization引用本文复制引用
李忠旺,陈玉梁,欧巧明,叶春雷,裴怀弟,刘新星,王红梅,罗俊杰..马铃薯病毒诱导应答基因抑制消减杂交文库构建及分析[J].草业学报,2017,26(12):152-159,8.基金项目
甘肃省农科院青年创新基金(2013GAAS29),甘肃省农业科学院创新团队(2015GAAS02),甘肃省农业科学院农业科技创新重大项目(2016GAAS59-02)和国家自然科学基金项目(31460388)资助. （2013GAAS29）
