油菜抗旱基因高通量克隆及功能分析

项目名称： 油菜抗旱基因高通量克隆及功能分析

项目编号： No.31470085

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 王道杰

作者单位： 河南大学

项目金额： 30万元

中文摘要： 油菜是我国重要的油料作物，同时又是耐旱性较差的作物，干旱严重制约其生产的发展。本课题组运用转录组测序（RNA-Seq）结合比较基因组学分析，筛选到68个可能参与油菜干旱胁迫响应的基因，并运用高通量的Gateway技术和FOX hunting system将68个基因全长cDNA转入拟南芥，获得包含378个转基因株系的油菜FOX拟南芥突变体库。本研究以这些油菜FOX植株为材料，运用高通量的远红外热成像技术筛选抗旱的油菜FOX突变株，结合抗旱表型、生理及细胞学分析，克隆获得相应的油菜干旱相关基因。通过基因表达分析并借助油菜近缘模式植物拟南芥干旱胁迫信号转导网络，发掘油菜干旱响应的关键节点基因。

中文关键词： 耐旱性；分子机制；基因克隆；功能分析；甘蓝型油菜

英文摘要： Rapeseed, as an important oil crop, is inclined to be subjected drought injury due to its relatively sensitive to drought stress among the primary crops. Using transcriptome sequencing (RNA-Seq) with comparative genomic analysis, 68 genes were screened out maybe involving in drought stress response in Brassica napus. these 68 genes were intrduced into Arabidopsis to construct rape FOX Arabidopsis mutant library by Gateway and FOX hunting system, including 378 FOX strains. In this study, take these rape FOX Arabidopsis mutants as materials, select drought-tolerant rape FOX transformants by high throughput far-infrared thermal imaging technology. Combining with drought-resistant phenotypic, physiological and cytological analysis, isolate candidate genes related drought stress from rape FOX Arabidopsis plants by PCR and sequencing. By gene expression analysis and according to the model plant Arabidopsis thaliana drought stress signal transduction networks, primarily explore key genes for drought stress responce in Brassica napus. Then use the genome editing techniques -CRISPR/Cas9 system and multi-gene over expression technology -MISSA system for the key genes functional verification in oilseed rape, preliminary elucidate the molecular mechanisms of these genes and collaborative regulatory mechanisms in drought-res

英文关键词： Drought resistance；Molecular mechanism；Gene cloning；Function analysis；Brassica napus L.