条锈菌诱导的小麦抑制差减杂交文库构建及其表达序列标签研究  

Construction of wheat SSH cDNA library induced by Puccinia striiformis and analysis of expressed sequence tags

作　　者：于秀梅[1] 喻修道[2] 屈志鹏[2] 韩青梅[3] 郭军[2] 黄丽丽[2] 康振生[2] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100 [2]西北农林科技大学植物保护学院,杨凌712100 [3]陕西省农业分子生物学重点实验室,杨凌712100

出　　处：《植物病理学报》2007年第1期50-55,共6页Acta Phytopathologica Sinica

基　　金：教育部重大科技培育项目(2004-295);教育部长江学者和创新团队发展计划资助项目(IRT0558);国家自然科学基金资助项目(30671350)

摘　　要：利用抑制差减杂交技术构建了条锈菌(Puccinia striiformis)诱导的小麦叶片cDNA文库。文库质量检测表明:差减杂交效率较高,质量较好。用DNAStar5.0对172条高质量的ESTs进行聚类,共获得120个contigs。对功能已知的contigs分类,能量和初级代谢类占32.5%,其中与光合作用相关的基因出现频率最高;参与膜及转运、抗病与防御的基因分别位于第二、三位;次生代谢、蛋白质合成加工和细胞结构建成的基因较少。通过分析抗病相关基因,初步推测HR和SAR可能为小麦抗条锈病过程中的2种抗性形式。最后利用RT-PCR对4条感兴趣基因进行分析,明确其在抗病过程中的表达模式。A suppression subtractive hybridization （SSH） cDNA library was constructed with wheat leaf induced by Puccinia striiformis race CY23. The process of library construction was checked and analyzed, results showed that the technique was efficient in the subtractive hybridization. 172 ESTs with good quality were assembled to 120 contigs with DNAStar 5.0 software. After function annotation, the most prevalent category was energy and primary metabolism which accounted for 32.5%, genes related with photosynthesis showed the highest frequency; the second category was membrane and transport, resistance and defense genes were in the third one;while genes involved in secondary metabolism, protein synthesis and process and cell structure were less. HR and SAR were supposed to be two main styles involving in resistance reaction through analyzing sequences with function identified genes. Four interested sequences were selected to testify their expression profile at different time after inoculating with P. striiformis race CY23 by reverse transcription PCR （RT-PCR）.

关 键 词：小麦 条锈菌 抑制差减杂交(SSH) 表达序列标签(ESTs) 反转录PCR(RT-PCR) 

分 类 号：S435.121.42[农业科学—农业昆虫与害虫防治]