摘要
为探究类黄酮3’-羟化酶(flavonoid 3’-hydroxylase,F3’H)基因在威氏绿绒蒿(Meconopsis wilsonii)花色形成过程中的表达规律。本研究以威氏绿绒蒿蓝紫色的花瓣为研究材料,使用RACE克隆技术克隆获得了威氏绿绒蒿花青素合成途径中的关键酶基因F3’H,命名为MwF3’H。分析发现其开放阅读框长度为1620 bp,编码了539个氨基酸,推测蛋白质分子量为59.7 kD,理论等电点(pI)为8.05,不稳定指数为33.02,属于稳定蛋白。序列比对和系统发育树分析显示,威氏绿绒蒿(M.wilsonii)与长果绿绒蒿(M.delavayi)的F3’H氨基酸序列一致性最高,相似度达到92.86%,聚为一个分支。实时荧光定量PCR结果显示,Mw F3’H基因在威氏绿绒蒿花发育不同的5个时期均有表达,在前期花青素积累时期表达最多,随着花的发育逐渐降低,但在盛花期表达量再次增长。推测其在威氏绿绒蒿花色的形成过程中起着重要作用。
To explored the regulation mechanism of flavonoid 3’-hydroxylase(F3’H)gene during the flower color formation of Meconopsis wilsonii.In this study,the key enzyme gene F3’H,named MwF3’H,in anthocyanin metabolism pathway was cloned from the blue-purple petals of M.wilsonii by using RACE cloning technology.Its open reading frame is 1620 bp and encoding 539 amino acids.The molecular weight of the protein is 59.7 kD and pI is 8.05,and the instability index is 33.02,which is a stable protein.Sequence alignment and phylogenetic tree analysis showed that the M.wilsonii and M.delavayi have the highest amino acid sequence identity with a similarity of 92.86%,and they are clustered into one branch.The results of real-time fluorescence quantitative PCR showed that the MwF3’H gene was expressed in five different periods of M.wilsonii flower development,and it was most expressed in the early accumulation period,and then gradually decreased with the development of flowers,but in the blooming flower,the expression increased again.It is speculated that it plays an important role in the formation of the flower color of M.wilsonii.
作者
严朋飞
张莹欣
贾维嘉
刘建
屈燕
Yan Pengfei;Zhang Yingxin;Jia Weijia;Liu Jian;Qu Yan(Provincial Key Laboratory of Ornamental Plants and Horticulture,College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming,650224;Southwest Landscape Architecture Engineering Research Center of State Forestry Administration,Southwest Forestry University,Kunming,650224)
出处
《分子植物育种》
CAS
北大核心
2022年第19期6382-6387,共6页
Molecular Plant Breeding
基金
国家自然科学基金项目(31460218)
云南省万人计划青年拔尖人才项目(YNWR-QNBJ-2019-211)
云南省教育厅科学研究基金项目(2020J0408)共同资助。
关键词
威氏绿绒蒿
类黄酮3’-羟化酶
花色
表达分析
Meconopsis wilsonii
Flavonoid 3’-hydroxylase
Flower color
Expression analysis
