灰霉病菌侵染后百合小分子RNA的识别及抗病分子机制的研究

项目名称： 灰霉病菌侵染后百合小分子RNA的识别及抗病分子机制的研究

项目编号： No.31460529

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 刘冠泽

作者单位： 云南农业大学

项目金额： 46万元

中文摘要： 云南作为我国百合重要的生产基地，灰霉病一直是威胁云南百合生产最主要的病害之一。发掘百合自身的抗性资源，利用现代生物学方法改良百合的防卫系统，是有效且环保的育种手段。而鉴定分离百合的抗病防卫相关基因，为培育广谱、持久抗性的百合新品种提供了新途径，也有助于推动百合抗病机理的不断发展。本研究通过对灰霉病菌侵染后的百合叶片进行转录组测序及小分子RNA测序，从转录水平及转录后调控水平，构建小分子RNA与灰霉病菌互作分子调控网络。并从中发掘出百合参与抗灰霉病菌的特异小分子RNA及其调控的靶标基因。然后，利用病毒介导的基因转录后沉默技术，完成候选小分子RNA及其靶标基因的高效筛选工作。结合分子生物学和基因工程手段，完成相关的小分子RNA和关键调控基因克隆和百合遗传转化，阐明百合中小分子RNA在灰霉病菌侵染过程的作用机制。同时，获得灰霉病的抗性提高百合育种材料，从而加速百合抗病的育种进程。

中文关键词： 百合；灰霉病；新一代测序技术；小分子RNA

英文摘要： Yunnan is the important production base of the lily in China. Grey mold disease caused by Botrytis is the most devastating diseases resulting heavy yield loss in Yunnan. It is an efficient and beneficial to the environment way of lily breeding that using the lily own genes to improve its defense system. Isolating and characterizing lily defense-responsive genes may provide new sources for improving the level of resistance of lily against pathogens and help to elucidate the mechanism of lily disease resistance. In this stuty, we employed the next generation high through sequencing technology to clone and identify both conserved and species-specific small RNAs and their targets in lily under the Botrytis cinerea. Combining the deep sequencing and bioinformatics, the study will build small RNA regulation and molecular mechanisms of Botrytis cinerea interaction network map from the level of transcriptional regulation post-transcriptional level. With the moleuclar biology and genetic engineering, we will isolating and cloning small RNA and their target genes involved in disease resistance. Using Virus-induced gene silencing(VIGS), the experiment complete efficiently screening of candidate target gene. Then, we plan to get the transgenic lily lines with overexpress of the candidatie target small RNA and key gene. Meanwhile, we will eluncidate the mechanism of small RNAs and their targets regulation of Botrytis cinerea resistance and develop lily lines which will accelerate the process resistance breeding of lily.

英文关键词： Lily;Grey mold disease;Next generation sequence;Small RNA