Identification and characterization of differentially expressed ESTs of Gossypium barbadense infected by Verticillium dahliae with suppression subtractive hybridization
Identification and characterization of differentially expressed ESTs of Gossypium barbadense infected by Verticillium dahliae with suppression subtractive hybridization
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DOI：
10.1007/s11008-005-0028-6
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发表时间：
2005-03
影响因子：
1.2
通讯作者：
K. Zuo;Jin Wang;Weisheng Wu;Y. Chai;Xiaofen Sun;K. Tang
中科院分区：
文献类型：
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作者：
K. Zuo;Jin Wang;Weisheng Wu;Y. Chai;Xiaofen Sun;K. Tang
棉花枯萎病防御反应是一个复杂的连续过程，涉及一系列基因。本研究采用抑制性消减杂交（SSH）技术，从海岛棉品种7124中分离黄萎病抗性过程中差异表达的EST。用反向北方印迹法对消减文库中的1165个克隆进行了筛选，其中131个EST被认为是过表达的，16个EST被认为是下调的。序列分析和blast搜索结果表明，83条EST与数据库中的45条独特序列同源。在所有这些差异表达的EST中，至少有三种基因被表征。大多数推定为有氧代谢酶的EST在感染过程中强烈表达。同样，在本研究中也挑选出了与病原体相关蛋白基因相似的EST。这些EST与其他激酶样基因和防御素样EST结合，构成了在病原体感染期间响应的基因集合。这些结果表明，海岛棉在受到病毒的攻击时，受到了强烈的氧化胁迫，并产生了一系列的防御反应。大丽花据我们所知，这是第一份关于海岛棉防御反应中全球ESTs分离的报告。
The wilt defense reaction of cotton is a complicated continuous process and involves a battery of genes. In this study, we adopted the suppression subtractive hybridization (SSH) technique to isolate differentially expressed ESTs fromGossypium barbadensevariety 7124 during theVerticilliumwilt defense process. An array of 1165 clones from the subtractive library has been screened with reverse northern blotting, of which 131 ESTs were considered as overexpressed and 16 ESTs were downregulated. Sequence analysis and blast search showed that 83 ESTs were homologous to 45 unique sequences in the databases. Among all these differentially expressed ESTs, at least three kinds of genes were characterized. The majority of ESTs with a deduced identity as aerobic metabolism enzymes were strongly expressed in the infection process. Likewise, ESTs similar to those reported for pathogen-related protein genes were also picked out in this study. These ESTs, in combination with other kinase-like genes and a defensin-like EST, constituted an assembly of genes which responded during pathogenic infection. These results imply that sea-island cotton undergoes strong oxidative stress and results in a series of defense responses when attacked byV. dahliae. To our knowledge, this is the first report on the isolation of global ESTs during the sea-island cotton defense reaction.