一个类黄酮3'-羟化酶参与多星韭花色素生物合成的功能解析  

Function analysis of a flavonoid 3'-hydroxylase gene involved in anthocyanidin biosynthesis of Allium wallichii

作　　者：孔德静 梁进丽 宫秋玲 俞沙沙 梁琳 孙威[2] 鞠志刚 KONG Dejing;LIANG Jinli;GONG Qiuling;YU Shasha;LIANG Lin;SUN Wei;JU Zhigang(Pharmacy College,Guizhou University of Traditional Chinese Medicine,Guiyang 550025;Key Laboratory of Plant Physiology and Development Regulation,School of Life Science,Guizhou Normal University,Guiyang 550025)

机构地区：[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

出　　处：《安徽农业大学学报》2024年第4期581-587,共7页Journal of Anhui Agricultural University

基　　金：贵州省教育厅特色领域项目(黔教合KY字[2021]059);贵州省“千层次”人才项目(贵中医[ZQ2018004]);贵州省高等学校高山杜鹃病虫害绿色防控重点实验室(黔教技[2022]044号);贵州省高等学校工程研究中心(黔教合KY字[2021]007);贵中医大创合字(2021)36号共同资助。

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,Aw F3'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。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 expressed in all tested tissues,displayed the highest transcript levels in the stamen and the lowest in the root.Compared 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 increased.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 

分 类 号：Q943.2[生物学—植物学]