百合黄烷酮3-羟化酶基因LhSorF3H的克隆与表达  

Cloning and Expression of LhSorF3H Genes in Lilium

作　　者：张星 杨捷[1] 彭梦笛 贾桂霞[1] 何恒斌[1] ZHANG Xing;YANG Jie;PENG Mengdi;JIA Guixia;HE Hengbin(Beijing Key Laboratory of Flower Germplasm Innovation and Molecular Breeding, National Flower Engineering and Technological Research Center, Beijing Laboratory of Urban and Rural Ecological Environment, Forest and Flower Breeding Laboratory of Ministry of Education, College of Landscape Architect)

机构地区：[1]花卉种质创新与分子育种北京市重点实验室,国家花卉工程技术研究中心,城乡生态环境北京实验室,教育部林木花卉育种实验室,北京林业大学园林学院,北京100083

出　　处：《西北植物学报》2017年第12期2325-2331,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31672191);中央高校基本科研业务费专项资金(YX2015-16)

摘　　要：为了探究百合花色的调控机制,以东方百合品种‘索邦’(Sorbonne)的花蕾为试验材料,根据前期转录组测序结果,成功克隆出黄烷酮3-羟化酶(flavanone 3-hydroxylase,F3H)基因的编码序列和基因组序列,其中,编码序列全长1 110bp,可编码369个氨基酸;基因组序列全长1 438bp,包含3个外显子和2个内含子,命名该基因为LhSorF3 H(GenBank登录号为MF614135)。生物信息学分析结果显示,黄烷酮3-羟化酶在进化上具有较高的保守性。LhSorF3 H编码的蛋白与郁金香(Tulipafosteriana)最为相似。半定量RT-PCR结果显示,LhSorF3H在花被、柱头以及花柱中表达明显,在花丝、子房、嫩茎、茎生根及上部叶片中表达较弱,而在花药和中下部叶片中基本不表达。LhSorF3 H在不同发育阶段的花被片中均有表达(S2和S6阶段除外);黑暗处理使LhSorF3 H表达降低,黑暗处理2h的LhSorF3 H表达量降到最低,之后表达量开始上升;转入光照条件后,LhSorF3H的表达水平持续增加。研究表明,LhSorF3 H基因对昼夜节律和光照具有双重响应的特性。The buds of oriental hybrid lily‘Sorbonne’were used to understand the regulatory mechanisms of flower coloration.Based on previous transcriptome database,we successfully cloned the coding sequence and genome sequence of flavanone3hydroxylase(F3H).The cDNA sequence had the encoding region of1110bp,and is predicted to encode a protein with369amino acids;the DNA full length of1438bp contained three exons and two introns,and is named LhSorF3H.Bioinformatics analysis results revealed that F3H was highly conservative in evolution and there is a high similarity between the proteins encoded by LhSorF3H and F3H of Tulipa fosteriana.Analysis by semi quantitative reverse transcription PCR(RT PCR)revealed that LhSorF3H gene showed the strongest expression in tepals,stigma and style,lower expression in filament,ovary,immature stem,stem root and upper leaf,the lowest expression in anther and lower leaf of‘Sorbonne’.LhSorF3H expressed in almost every stage except S2and S6.In addition,dark treatment can reduce the expression of LhSorF3H,and it showed a lowest expression after two hour dark treatment,and after that,the expression started to go up.With the increasing of illumination time,the expression level of LhSorF3H began to increase slightly,but there was no significant difference between the light time groups.

关 键 词：百合'索邦’ 黄烷酮3羟化酶基因 克隆 表达 

分 类 号：Q785[生物学—分子生物学] Q786