黄萎病菌诱导海岛棉全长cDNA文库构建及EST分析  

Construction of A Full-length cDNA Library Inoculated by Verticillium dahliae in Gossypium barbadense and Analysis of the EST

作　　者：张艳[1] 丁泽国 王省芬[1] 张桂荣[1] 李志坤[1] 张桂寅[1] 吴立强[1] 马峙英[1] 

机构地区：[1]河北农业大学/教育部华北作物种质资源研究与利用重点实验室,河北保定071001

出　　处：《棉花学报》2014年第3期274-278,共5页Cotton Science

基　　金：国家自然科学基金(31301371);国家863子课题(2013AA102601-5)

摘　　要：采用SMART技术构建了海岛棉黄萎病菌诱导下根全长cDNA文库,并进行了质量鉴定和EST序列分析。结果表明,文库的重组率为92.3%,库容量为1.1×106pfu·mL-1,平均插入片段大于1.5 kb。生物信息学分析表明该文库包含大量抗病调节基因和抗病防御基因。COG功能分类发现了多个参与信号转导、防御反应、细胞骨架以及次生代谢产物合成与分解等不同功能类别的基因成员。文库中出现的高丰度表达基因主要有病程相关蛋白(PR蛋白)、谷胱甘肽硫转移酶基因(GST)、亲环素蛋白、短链乙醇脱氢酶、抗坏血酸过氧化物酶及转录因子等。文库的构建为深入研究棉花抗黄萎病分子机制奠定基础。A full-length cDNA library inoculated by Verticillium dahliae was constructed with the Gossypium barbadense cv.Pima 90-53 roots based on SMART technique,the quality evaluatation and EST analysis of this library were finished.The recombinant rate of present library was 92.3％,the titer was 1.1 × 106 pfu·mL-1,and the average size of inserted eDNA fragments was about 1.5 kb.Bioinformatics analysis of EST data indicated that a lot ofunigenes with homology to known defense-related genes in other plants were found in our library.Clusters of orthologous group（COG） functional classification found that more genes which involved in signal transduction,defense mechanisms,cytoskeleton and secondary metabolites biosynthesis/transport/catabolism were detected.And the highly-expressed genes were detected,which mainly contained pathogenesis-related proteins（PR proteins）,glutathione-S-transferase（GST）,cyclophilin,short chain alcohol dehydrogenase,ascorbic acid peroxidase,transcription factor,and so on.These will be of significance to understanding the molecular mechanism of cotton Verticillium wilt resistance.

关 键 词：海岛棉 黄萎病 CDNA文库 EST分析 

分 类 号：S562[农业科学—作物学]