Function analysis of a flavonoid 3'-hydroxylase gene involved in anthocyanidin biosynthesis of Allium wallichii
孔德静 1梁进丽 1宫秋玲 1俞沙沙 1梁琳 1孙威 2鞠志刚1
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作者信息
1. 贵州中医药大学药学院,贵阳 550025
2. 贵州师范大学生命科学学院/植物生理与发育调控重点实验室,贵阳 550025
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摘要
类黄酮3'-羟化酶(F3'H)在花色素苷生物合成过程中起关键作用.明确多星韭F3'H在花色素苷生物合成中的功能,将为多星韭花色形成及改良研究提供基因资源.基于转录组数据从多星韭花朵中克隆获得AwF3'H,并对其进行生物信息学分析,利用Real time PCR对AwF3'H1的表达模式进行分析,并利用蘸花法将AwF3'H转入拟南芥中,同时对转基因植株进行表型观察及花色素苷检测分析.结果表明,AwF3'H的ORF全长为1545 bp,编码514个氨基酸,属于细胞色素P450家族成员,与洋葱AcF3'H的亲缘关系最近;在所检测组织均有表达,但在雄蕊中表达最高,根中表达最低;与野生型拟南芥相比,转AwF3'H拟南芥T2代幼苗子叶及下胚轴颜色明显加深,矢车菊素与天竺葵素类花色苷积累量显著增加.表明AwF3'H具有类黄酮3'-羟化酶的功能.
Abstract
Flavonoid 3'-hydroxylase (F3'H) plays a key role during anthocyanin biosynthesis.To elucidate the function of F3' H in anthocyanin biosynthesis will provide gene resources for the flower color formation and improvement of Allium wallichii.AwF3'H was cloned from A.wallichii flower based on transcriptome data and analyzed by bioinformatics.The expression pattern of AwF3'H was examined by Real time PCR.Subsequently,AwF3'H was transferred into Arabidopsis by flower dipping method,and the phenotype as well as anthocyanin of transgenic plants were identified.The results showed that the ORF of AwF3'H was 1545 bp,encoding 514 amino acids.It is a member of cytochrome P450 family and is closely related to AcF3'H of onion.AwF3'H was ex-pressed in all tested tissues,displayed the highest transcript levels in the stamen and the lowest in the root.Com-pared with wild type Arabidopsis,the color of cotyledon and hypocotyl of T2 Arabidopsis seedlings containing AwF3'H was significantly darker,cyanidin and pelargonidin anthocyanins accumulation was dramatically in-creased.The results indicated that AwF3'H functions as a flavonoid 3'-hydroxylase.
关键词
多星韭/基因功能验证/类黄酮3'-羟化酶/表达分析/花青素
Key words
Allium wallichii/gene function verification/flavonoid 3'-hydroxylase/expression analysis/anthocyanidin
万方数据