菘蓝中靛蓝生物合成关键酶基因CYP的鉴定与功能研究

项目名称： 菘蓝中靛蓝生物合成关键酶基因CYP的鉴定与功能研究

项目编号： No.31200221

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 微生物学、植物学

项目作者： 李焘

作者单位： 陕西师范大学

项目金额： 23万元

中文摘要： 靛蓝是菘蓝的主要代谢产物，其基本合成过程为：吲哚→吲哚酚→靛蓝前体→靛蓝。其中，吲哚在细胞色素P450单加氧酶的催化下加羟基形成吲哚酚，是吲哚形成靛蓝的重要分支过程。据此，我们推测编码细胞色素P450单加氧酶的CYP基因在靛蓝合成过程中具有重要作用。研究前期采用Solexa高通量测序技术进行菘蓝转录组de novo测序，共获得51,698 条Unigene，其中靛蓝合成相关基因23条，CYP基因4条。本研究拟通过分析生物（真菌诱导子）及非生物胁迫（光照、干旱、盐等）条件下CYP基因的表达变化与靛蓝积累之间的关系，筛选并克隆与靛蓝合成相关的CYP基因家族成员，进而研究其时空表达特性；采用过表达及基因敲除的方法，进一步阐明CYP基因在靛蓝生物合成过程中的作用。本项目预期研究结果将为深入探讨靛蓝生物合成的分子机制，利用生物技术生产靛蓝提供新的思路和策略。

中文关键词： 菘蓝；CYP基因；转录组测序；次生代谢产物；

英文摘要： Indigo is the main secondary metabolites of Isatisindigotica Fort. The basic biosynthesis process of indigo is: indole→indoxyl→precursors of indigo→indigo. Among them, the CYP (cytochrome P450) is the key enzyme which oxidized indole to indoxyl by adding hydroxyl groups. This process is the branch point of synthesis of indigo in plant. By using Solexa deep sequencing technology, we obtained 51,698 unigenes from transcriptome of Isatis indigotica Fort..There are 23 enzymes participated in indigo biosynthesis, including 4 CYP genes which involved in hydroxylation of indole. In this proposal, we intend to analyze the relationship between the change of CYP gene expression and the accumulation of indigo in vivo under the biotic stress (fungal elicitors) and abiotic stress (light, drought, salt, etc.) conditions. By screening and cloning CYP genes, we also consider the characteristics of spatio-temporal expression of the CYP genes in vivo and confirm the functions of CYP gene in regulation of indigo synthesis process. In this study, we attempt to clarify the molecular mechanisms of indigo biosynthesis, and try to open up a new ways to produce indigo by bio-technology. Indigo is the main secondary metabolites of Isatisindigotica Fort. The basic synthesis process of indigo is: indole?indoxyl?precursors of indigo?indigo.

英文关键词： Isatis indigotica Fort.；CYP genes；transcriptome sequencing；secondary metabolite；