苹果树腐烂病菌致病木聚糖酶的基因功能及其抑制蛋白的鉴定

项目名称： 苹果树腐烂病菌致病木聚糖酶的基因功能及其抑制蛋白的鉴定

项目编号： No.31272001

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 王彩霞

作者单位： 青岛农业大学

项目金额： 75万元

中文摘要： 黑腐皮壳菌引起的苹果树腐烂病是对苹果产业威胁最大的一种毁灭性病害，但迄今对腐烂病菌的致病因子及其致病机理尚不明确，导致现有防治措施不能有效控制病害的发生和流行。本课题组研究证实腐烂病菌侵染致病过程中产生活性极强的木聚糖酶，且酶活性与菌株致病力存在密切相关性；在研究中还发现寄主的抗病性与其抑制木聚糖酶活性的能力呈正相关。本项目拟从苹果树腐烂病菌分泌的木聚糖酶入手，通过构建基因组文库和RACE技术，分离克隆木聚糖酶同源基因的全长序列；利用基因表达分析和基因敲除技术，解析木聚糖酶基因的功能；以此为基础，筛选致病木聚糖酶的抑制蛋白，分析其在植物抗病防御反应中的作用，进而研究抑制蛋白对腐烂病菌生长、繁殖及侵染的影响。研究结果将明确腐烂病菌的致病因子及木聚糖酶基因在病原菌致病性中的作用，明确苹果木聚糖酶抑制蛋白对腐烂病的生防效果，为全面揭示腐烂病菌的致病机理及应用酶抑制蛋白防治植物病害奠定基础。

中文关键词： 苹果树腐烂病菌；木聚糖酶；基因表达；基因敲除；抑制蛋白

英文摘要： Apple tree canker, caused by Valsa ceratosperma, is one of the most devastating and economically important diseases of apple prcduction in China. At present, measures to control losses caused by V. ceratosperma include using agriculture management cultures, chemical protection and treatment diseased scars. However, these measures can not control the disease effectively because the pathogenic factors and pathogenicity mechanisms of V. ceratosperma is still not clear. Our research group had confirmed that V. ceratosperma could produce a series of cell wall-degrading enzymes (CWDEs) including xylanase, polygalacturonase (PG), polymethylgalacturonase ( PMG), cellulose (Cx) and β-glucosidase. In all the CWDEs, the maximum activity of xylanase was the highest and it showed the most powerful macerating ability. The enzyme activities of xylanase were closely correlated with the virulence of different strains. In additon, it was showed that the disease resistance of plant and it's the inhibiting ability to xylanase activity was positive correlation, which provides a new idea for the control of apple valsa canker. This proposed project will investigate three aspects of the problem starting from xylanase produced by V. ceratosperma in the infection process. The first aspect is to isolate and clone the genes of xylanase by

英文关键词： Valsa mali var. mali；xylanase；gene expression；gene knockout；xylanase inhibitor