不结球白菜耐寒关键基因的鉴定、分析及功能研究

项目名称： 不结球白菜耐寒关键基因的鉴定、分析及功能研究

项目编号： No.31330067

项目类型： 重点项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 侯喜林

作者单位： 南京农业大学

项目金额： 288万元

中文摘要： 低温是限制作物产量和品质的一个重要因素。不结球白菜是我国重要的蔬菜作物之一，但经常遭受低温甚至冻害的影响，使其产量和品质下降。本研究在拥有不结球白菜五个变种1986份资源的基础上，已完成102份材料的重测序（其中耐寒资源24份），还从耐-9.7℃的材料中克隆到了3个耐寒基因（BrICE1、BrCBF和BrCOR14）；拟通过对不结球白菜耐寒资源和不耐寒资源进行简化基因组测序，利用全基因组关联分析方法，发掘耐寒基因，并对其编码的蛋白质进行生物信息学分析。利用转录组测序、酵母双杂交、蛋白质免疫共沉淀和染色质免疫共沉淀等技术，研究耐寒基因及转录因子间的互作机理以及调控通路网络。然后利用基因过量表达、TILLING、VIGS等技术对抗寒相关基因进行系统的功能验证，最终确定耐寒关键基因。本研究探索出的耐寒基因遗传研究方法，可为不结球白菜乃至其它作物耐寒新品种的选育提供理论依据和技术手段。

中文关键词： 不结球白菜;耐寒基因;全基因组关联分析;功能研究;网络调控

英文摘要： The low temperature is an important factor limiting crop yield and quality. Non-heading Chinese cabbage is one of the important vegetable crops in China. But low temperatures and even freezing temperatures often affect its yield and quality. Based on the 1,986 germplasm resources of five variants of non- heading Chinese cabbage, we have resequenced 102 key germplasms of non-heading Chinese cabbage (24 cold-resistant cultivars) and cloned 3 cold-resistant genes from the cultivar that can survive at -9.7℃ environment. In this study we will screen out key cold-resistant genes or genetics sites using GWAS (Genome-wide Association Study) after reduced-representation sequencing database of cold-resistant and non-cold-resistant non-heading Chinese cabbage. Then we will analyze their gene structure, and gene transcription and expression profiling to explore regulation network of cold-resistance by RNA sequencing, two yeast hybrid, protein and chromatin co-immunoprecipitation. Furthermore, we will verify the function of cold-resistant genes by gene overexpression, VIGS and TILLING. The genetics study methods of cold-resistant genes with bioinformatics will be helpful to molecular breeding for cold-resistant non-heading Chinese cabbage or other crops.

英文关键词： non-heading Chinese cabbage;cold-resistant genes;Genome-wide Association Study;functional study ;regulation network