调控黄花龙胆玉米黄质环氧化酶基因表达的转录因子基因的克隆和功能分析

项目名称： 调控黄花龙胆玉米黄质环氧化酶基因表达的转录因子基因的克隆和功能分析

项目编号： No.31270344

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 朱长甫

作者单位： 长春师范大学

项目金额： 92万元

中文摘要： 类胡萝卜素对人类营养和健康具有重要价值。在应用遗传工程技术提高植物靶器官中某一特定类胡萝卜素含量时，存在很大的盲目性，这主要是由于目前人们对植物体内类胡萝卜素生物合成基因表达调控的分子机制以及类胡萝卜素积累的调控机制知之甚少。研究表明黄花龙胆玉米黄质环氧化酶(ZEP)基因表达与类胡萝卜素合成以及黄花龙胆花瓣和番茄果实有色体发育紧密相关。本项目拟用酵母单杂交和番茄果实瞬时表达技术从黄花龙胆花瓣中克隆和鉴定调控ZEP基因表达的转录因子基因；观测在番茄中稳定超表达转录因子对果实等器官内源类胡萝卜素基因表达和类胡萝卜素组分和含量的影响；用酵母双杂交等技术克隆和鉴定编码与转录因子相互作用的相关蛋白质基因；揭示他们在番茄果实有色体分化中对类胡萝卜素合成基因表达和类胡萝卜素积累的调控机制，为利用基因工程技术在含有有色体的花和果实等器官中生产类胡萝卜素以及改变植物花颜色的分子育种提供新的基因源和奠定基础。

中文关键词： 龙胆；玉米黄质环氧化酶；启动子；转录因子；花青素苷

英文摘要： Carotenoids play a critical role in human nutrition and health. There have been many examples of unintented/unpredicted phenotypes and results in metabolic enginnering of carotenoids in higher plants. Typically this has resided from our lack of understanding of molecular regulation of carotenogenic gene expression and carotenoid formation in planta. Zeaxanthin epoxidase (ZEP) catalyzes the conversion of zeaxanthin to violaxanthin via antheraxanthin, and is the key enzyme responsible for the accumulation of antheraxanthin and violaxanthin in gentian (Gentiana lutea) flower petals. It has been demonstrated that the expression of gentian (G. lutea) zeaxanthin epoxidase (GlZEP) gene is strictly associated with carotenoid formation and chromoplast differentiation in gentian flowers and tomato fruits. In this project, the cDNAs encoding transcription factors for regulation of GlZEP gene expression will be isolated from cDNA library of gentian flower petals using Yeast One-Hybrid (Y1H) system, and characterized using transient expression system in tomato fruits by Agroinjection. The stable transgenic tomato plants over-expressing the putative transcription factor gene will be analyzed simultaneously for endogenous carotenogenic gene expression and carotenoid profile in different organs, flowers and fruits at different

英文关键词： Gentian；zeaxanthin epoxidase；promoter；transcription factor；anthocyanins