小麦与条锈菌亲和互作的差减文库构建及初步分析  

Construction and Preliminary Analysis of the Compatible SSH cDNA Library Between Puccinia striiformis and Wheat

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

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

出　　处：《中国农业科学》2008年第5期1267-1273,共7页Scientia Agricultura Sinica

基　　金：国家“973”前期专项(2006CB708208);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);高等学校学科创新引智计划资助项目(B07049)

摘　　要：【目的】构建小麦与条锈菌亲和互作的差减文库,分离条锈菌侵染小麦过程中的差异表达基因。【方法】以小麦品种水源11和条锈菌相应毒性小种CY31号为材料,构建条锈菌侵染阶段的SSH-cDNA文库,挑选250个阳性克隆测序并进行生物信息学分析。【结果】聚类后得到149条非冗余EST(unigene)。经BlastX比对和功能分类分析,其中50条unigene(33.6%)未找到同源性匹配,25条(16.8%)与未知功能蛋白同源性较高;其余74条功能已知的unigene中,与初级代谢、能量相关的基因分别有13条和10个,占8.7%和6.7%,感病及防御相关的基因有6个约占4.0%;此外,获得两个与病原菌有较高同源性的基因。随后,进一步利用RT-PCR对6个基因的表达模式进行了分析。【结论】成功构建了小麦与条锈菌亲和互作的差减文库,分离出一部分与小麦条锈病发病相关的基因,可用于进一步研究条锈菌侵染过程中特异基因的功能。[Objective] This study was conducted to isolate differentially expressed genes during the compatible interactions between Puccinia striiformis and wheat. [Method] A SSH-cDNA library was constructed with wheat variety Shuiyuan-11 and a Chinese virulent race CY31 of Puccinia striiformis, 250 positive clones were selected for sequencing. [Result] One hundred and fourty-nine nonredundant ESTs were assembled with CAP3 in total, and then function annotation and classification were performed. There were 50 unigenes （33.6%） without or with weak similarities to the existent proteins in the nr database from NCBI and 25 unigenes （16.8%） significantly matched with hypothetical protein. While the unigenes assigned to primary metabolism, energy and disease/defence accounted for 8.7%, 6.7%, and 4.0%, respectively. Two unigenes, however, had higher similarity with pathogenic protein. Six genes were selected to investigate their expression profiles by reverse transcription PCR （RT-PCR）. [Conclusion] This high quality SSH cDNA library can be further used for gene cloning and provide a foundation for further investigation to explore the molecular mechanism of interaction between wheat and stripe rust fungtis.

关 键 词：小麦 条锈菌 抑制差减杂交 亲和互作 反转录PCR 

分 类 号：S435.121[农业科学—农业昆虫与害虫防治]