甜荞花青素糖基转移酶基因FeUFGT1的克隆、遗传转化及生物信息学分析  

Cloning,Genetic Transformation and Bioinformatics Analysis of the Glycosyltransferase Gene FeUFGT1 of Common Buckwheat

作　　者：张兰 杨铝 杨朝结 陈红 黄娟 朱丽伟 陈庆富[1] 邓娇 ZHANG Lan;YANG Lü;YANG Chaojie;CHEN Hong;HUANG Juan;ZHU Liwei;CHEN Qingfu;DENG Jiao(Guizhou Buckwheat Engineering Technology Research Center,College of Life Sciences,Guizhou Normal University,Guiyang 550025,China)

机构地区：[1]贵州师范大学生命科学学院,贵州省荞麦工程技术研究中心,贵州贵阳550025

出　　处：《华北农学报》2024年第3期43-50,共8页Acta Agriculturae Boreali-Sinica

基　　金：贵州省科技计划项目(黔科合基础-ZK[2023]一般278);国家自然科学基金青年科学基金项目(32102422);国家现代农业产业技术体系荞麦育种岗位科学家专项资金(CARS-07-A5);国家自然科学基金项目(32060508)。

摘　　要：为探究花青素糖基转移酶基因(UFGT)在甜荞花色苷生物合成中的作用,以甜荞贵红1号为材料,基于同源比对分析,筛选并克隆得到1条UFGT基因序列,命名为FeUFGT1。对FeUFGT1进行生物信息学分析,并构建过表达载体转入拟南芥花青素3-O-糖基转移酶突变体(atugt78d2)中分析其功能。结果表明,FeUFGT1开放阅读框为1 404 bp,编码467个氨基酸残基,推测其分子质量51.46 ku,理论等电点为4.97。FeUFGT1蛋白不存在信号肽和跨膜区,含有植物糖基转移酶家族典型结构域(GT-B型),并且在C末端具有植物UGT家族成员特有的PSPG-box。进化分析表明,FeUFGT1蛋白与罗汉果的3-O-葡萄糖基转移酶UGT74AC1亲缘关系最近。在白花品种丰甜1号和红花品种贵红1号2个品种盛花时期的花瓣中,红花中FeUFGT1的表达量约为白花中的3.7倍,差异显著。突变体恢复试验表明,FeUFGT1能够恢复拟南芥突变体atugt78d2缺乏花色苷积累的表型,具有催化花色苷生成的功能。This study aimed to investigate the function of the anthocyanidin glycosyltransferase gene(UFGT)in the anthocyanin biosynthesis in common buckwheat.Guihong 1 was used as the experimental material.A UFGT gene,named FeUFGT1,was obtained by homologous alignment analysis,and cloned followed by performing on bioinformatics analysis.Overexpression vector of this gene was constructed,then transferred into Arabidopsis thaliana anthocyanin 3-O-glycosyltransferase mutant atugt78d2 to assess its function.The results revealed that the open reading frame of FeUFGT1 was 1404 bp and encoded 467 amino acid residues with speculated molecular weight of 51.46 ku and theoretical isoelectric point of 4.97.FeUFGT1 protein contained GT-B type glycosyltransferase family domain and a PSPG-box,specific one of the plant UGT family at the C-terminus.The phylogenetic analysis showed that FeUFGT1 protein was closely related to the 3-O-glucosyltransferase of Siraitia grosvenorii UGT74AC1.The expression level of FeUFGT1 in the white-flower petals of Fengtian 1 was 3.7-fold as high as that in the red-flower petals of Guihong 1 with a significant difference.Mutant recovery experiment showed that FeUFGT1 could restore the phenotype of the Arabidopsis mutant which lacked anthocyanin accumulation.

关 键 词：甜荞 FeUFGT1 基因克隆 花色苷 3-O-葡萄糖基转移酶 

分 类 号：S517.03[农业科学—作物学] Q78[生物学—分子生物学]