联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月5日 星期六
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
葡萄风信子二氢黄酮醇 4- 还原酶基因(DFR)的克隆与表达分析
焦淑珍1,刘雅莉2,娄倩1,姜玲
Cloning and Expression Analysis of Dihydroflavonol 4-reductase Gene(DFR) from Grape Hyacinth(Muscari armeniacum)
全文: PDF (1863 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 葡萄风信子(Muscari armeniacum)是一类重要的球根花卉,其花色呈现特殊的蓝色。花青素是影响葡萄风信子蓝色形成的重要物质,与植物生长过程及人类/动物饮食密切相关。有关花青素生物合成途径的研究已较为成熟,但由于葡萄风信子基因资源缺乏,其花色形成的机理目前尚不清楚。二氢黄酮醇 4- 还原酶(dihydroflavonol 4-reductase, DFR)是植物花青素合成中的重要酶。本研究采用RACE技术从葡萄风信子花瓣中克隆到两条DFR基因(MaDFR2a和MaDFR2b),GenBank登录号分别为KJ619963和KJ619964。序列分析表明,MaDFR2a和MaDFR2b全长1 392 bp,包含一个长度为76 bp的5'非翻译区和215 bp的3'非翻译区,均包括一个1 101 bp的完整开放阅读框(open reading frame, ORF),编码366个氨基酸,氨基酸相似性为98%,与其他植物的DFR蛋白同源性较高。聚类分析表明,葡萄风信子DFR最先与单子叶植物聚为一类。生物信息学分析表明,两个DFR蛋白可能为亲水性蛋白,均具有两个跨膜区,无信号肽。亚细胞定位分析显示,两个DFR基因均定位于整个细胞中。α-螺旋和无规则卷曲是两个DFR蛋白的主要二级结构元件。MaDFR2a和MaDFR2b编码的蛋白三级结构非常相似,中间形成一个裂沟,可以进行催化反应。花青素含量测定显示,着色的花器官中花青素含量较高,根、茎、叶以及未着色的花蕾中几乎检测不到花青素。荧光定量PCR分析发现,MaDFR2a和MaDFR2b在葡萄风信子花组织中表达较高,根、茎和叶中微量表达;MaDFR2a和MaDFR2b均在未着色的花蕾时期开始表达,随着花色加深,在完全着色的花中表达最高,此后逐渐降低,但MaDFR2b在未着色的花蕾期和完全着色的花中表达相对较高。研究结果表明,DFR可能是影响蓝色葡萄风信子形成的重要基因。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
焦淑珍
刘雅莉
娄倩
姜玲
关键词 花色葡萄风信子DFR克隆表达分析    
Abstract:Grape hyacinth (Muscari armeniacum) is an important ornamental bulbous plant with increasing economic, ornamental and scientific importance because of their outstanding blue color. Anthocyanins are principal flower pigments in M. armeniacum, and indispensible in plant biology and human/animal diets. Whereas anthocyanin synthesis is well characterized in numerous plants, the mechanism underlying the blue appearance of M. armeniacum is still far from understanding for the little knowledge of gene information of this plant. Here, two dihydroflavonol 4-reductase (DFR) genes, which encode a later enzyme for anthocyanin formation, were isolated from Muscari armeniacum petals using RACE techniques and designated as MaDFR2a and MaDFR2b(GenBank accession No. KJ619963 and KJ619964), respectively. Sequence analysis of cDNAs revealed that the full length sequence was 1 395 bp, containing a length of 76 bp 5' untranslated region and 215 bp 3' untranslated region,both of them contained a 1 101 bp open reading frame (ORF), which encoded a protein of 366 amino acids. The similarity of the two DFR was 98%. Homology analysis showed that the deduced DFR proteins were highly homologous to other DFR proteins from different plant species. The cluster analysis results showed that the two DFR genes clustered together with monocotyledons firstly. Bioinformatics showed that two DFR proteins were both hydrophilic proteins, containing two transmembrane domains and neither of them had signal peptide. Subcellular localization analysis showed that two DFR genes located in the whole cells. α-Helix and random coil were primary secondary structural components of the two DFR genes. The tertiary structures of two DFR proteins were similar. Both of them had a fissure which could do some catalytic reactions. High performance liquid chromatography (HPLC) analysis showed that high contents of color anthocyanins were detected in pigmented flower organs, while no anthocyanins were detected in root, stem, leaf and unpigmented flower buds. The result of Real-time fluorescence quantitative PCR analysis demonstrated that the DFRs expression was presented in flowers but little or very weak expression was also detected in the other tissues (leaf, stem and root). When flower pigmentation started, a steep rise of two DFR genes expression occurred and peaked at fully coloured flowers. Subsequently a slight decrease in the MaDFR2a and MaDFR2b expression was presented at later stage. In contrast, a relatively high-level of MaDFR2b expression was found in both unpigmented or fully-pigmented buds. This result suggested that DFR can play a role in the color formation of blue grape hyacinth.
Key wordsFlower color    Muscari armeniacum    DFR    Cloning    Expression analysis
收稿日期: 2013-10-21     
通讯作者: 刘雅莉   
引用本文:   
焦淑珍1,刘雅莉2,娄倩1,姜玲. 葡萄风信子二氢黄酮醇 4- 还原酶基因(DFR)的克隆与表达分析[J]. , 2014, 22(5): 529-540.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I5/529
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn