马铃薯糖苷生物碱合成的分子调控研究

项目名称： 马铃薯糖苷生物碱合成的分子调控研究

项目编号： No.31260343

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 张金文

作者单位： 甘肃农业大学

项目金额： 54万元

中文摘要： 马铃薯糖苷生物碱（GAs）是抗虫、抗病原体并具有渗透调节功能的重要成分，但高浓度GAs对人体有毒。开展马铃薯GAs合成和分布的空间遗传调控研究，对于探索次生代谢物分子调控策略和方法、提高植物抗逆性和改良品质将具有重要的理论和实践意义。为了精确地控制马铃薯不同器官中GA的合成和积累，本项目拟采用分子克隆技术分离GAs合成代谢途径的关键酶基因及叶片和块茎特异性启动子，分别构建叶片特异表达和块茎特异沉默载体，转化模式植物或马铃薯以获得转基因材料；采用实时定量PCR技术和HPLC方法鉴定转化材料中不同器官和组织中关键酶基因的表达水平和GAs含量，以此为依据筛选出具有器官高度特异性启动子及其有效的基因表达载体结构，最终构建能够使GAs合成的关键酶基因在叶片中过表达和块茎中受到抑制的整合载体，通过农杆菌介导法获得叶片GAs含量丰富而食用块茎含量低的材料，为马铃薯育种创新抗逆种质资源开辟一条新的途径。

中文关键词： 糖苷生物碱；代谢调控；糖基转移酶；基因表达；马铃薯

英文摘要： The potato (Solanum tuberosum L.) glycoalkaloids (GAs) are important components of plant resistance against pathogens and pests as well as osmotic adjustment but can be toxic to humans at high levels. The study of space genetic regulation of potato GAs biosynthesis and distribution will has important theoretical and practical significance for exploring molecular regulation strategies and methods of plant secondary metabolite, improving resistance to biotic or abiotic-stress and quality of plant. In order to accurately control the GAs biosynthesis and accumulation in different organs of potato, the project will clone the key enzyme genes associated with GAs biosynthesis and leaf-specific and tuber-specific promoters by molecular techniques, construct vectors that have efficient leaf-specific expression of the target gene and tuber-specific inhibited expression respectively, and gain genetically modified material by transforming model plant or potato. We will analyze the target-gene expression levels by real-time quantitative polymerase chain reaction (RQ-PCR) method, meanwhile determine the GAs content by HPLC method in different organs of the transformation materials and according to the results we will screen out the highly organ-specific promoters and their effective gene expression vector structure.Eventually

英文关键词： Glycoalkaloids；Metabolic regulation；Glycosyltransferase；Gene expression；Solanum tubersum