甜菜种质资源抗旱相关基因的发掘及功能分析

Sugar beet is one of the most important sugar crops in our country. Sugar beet is mainly distributed in northeast, northwest and north of China.At the same time, it is also an important industrial crops with regional characteristics and dvantages in Inner Mongolia for a long time. Drought is an important affecting factor for sugar beet production. We want to establish a rapid and convenient technology system to discovery drought tolerant genes resources in sugar beet, through the studies in the lab and field. Through the studies in the project we also want to construct a cDNA library of sugar beet induced by drought stress based on SMART technique, to get cDNA sequences with high througthput sequencing Technology, and excavate drought-resistance functional genomics information through high-performance bioinformatics technology. The gene will be cloned and its expression products will be detected and analysed to determine the control genes and after be transformed in Arabidopsis. Thus the project will provide gene resources for the research of gene engineering of drought resistant sugar beet and lay a foundation for further research on molecular mechanisms of drought resistance sugar beet.

甜菜是我国主要糖料作物之一，其种植区域主要分布在我国的东北、西北和华北地区，长期以来，甜菜是内蒙古自治区具有地区特色和优势的重要经济作物。目前，干旱已成为发展甜菜生产规模和提高甜菜产量的重要限制因素。本项目通过室内、田间相结合的方法对甜菜种植资源抗旱性进行综合评价与鉴定，建立一套快速、简便发掘甜菜抗旱基因资源的技术体系；采用SMART技术构建甜菜干旱胁迫诱导cDNA文库，通过高通量测序技术平台获得大量cDNA序列，通过高性能生物信息学平台挖掘甜菜抗旱功能基因组信息，克隆抗旱相关基因的全长cDNA进行表达产物分析，确定目标功能基因，并进行转基因模式植物的功能验证。为甜菜抗旱基因工程研究提供基因资源，也为阐明甜菜抗旱分子机制奠定理论基础。

干旱是制约甜菜生产发展和产质量提高的重要因素。本课题通过室内、田间相结合的方法对甜菜种质资源抗旱性进行了综合评价，建立了快速、简便发掘甜菜抗旱资源的技术体系，并获得不同抗旱性的甜菜种质资源；通过不同抗旱性甜菜对水分胁迫的响应研究，阐明了甜菜抗旱的生理基础；通过转录组测序（RNA-seq）技术构建了甜菜抗旱差异基因的表达模式；通过双向电泳分离水分胁迫下差异表达蛋白质并质谱分析后，获得了可能参与甜菜抗旱调控的17个显著差异表达蛋白点；基于转录组并结合生物信息学分析筛选了3个抗旱相关基因，构建了3个抗旱相关基因的植物表达载体，并分别将其转化拟南芥野生型，筛选获得各自稳定表达的转基因T3代阳性材料并进行了抗旱性功能鉴定，表明3个基因均可在一定程度上增强植物抗旱性。同时还研发了甜菜壮苗剂并应用于甜菜生产。