牡丹WD40类转录因子基因PsWD40-1和PsWD40-2的分离与序列分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (2): 85-90.

牡丹WD40类转录因子基因PsWD40-1和PsWD40-2的分离与序列分析

张超, 高树林, 杜丹妮, 吴凡, 董丽

北京林业大学园林学院国家花卉工程技术研究中心，北京 100083

收稿日期:2013-09-09 出版日期:2014-02-27 发布日期:2014-02-27
作者简介:张超，男，博士研究生，研究方向：牡丹切花花色发育；E-mail：zhangchao24804@gmail.com
基金资助:
高等学校博士学科点专项科研基金（20130014110014）

Isolation and Sequence Analysis of the Paeonia suffruticosa WD40 Transcription Factor Genes PsWD40-1 and PsWD40-2

Zhang Chao, Gao Shulin, Du Danni, Wu Fan, Dong Li

（College of Landscape Architecture，Beijing Forestry University，National Flower Engineering Technology
Research Center，Beijing 100083）

Received:2013-09-09 Published:2014-02-27 Online:2014-02-27

摘要/Abstract

摘要： 利用已构建的牡丹‘洛阳红’花瓣转录组数据库，筛选得到2个与植物花青素苷合成WD40蛋白同源性较高的Unigene序列，分别命名为PsWD40-1和PsWD40-2。利用RT-PCR技术，设计特异性引物对PsWD40-1和PsWD40-2最大阅读框（ORF）序列进行扩增，测序结果表明PsWD40-1序列包含一个1 035 bp的ORF，编码一个344 aa的肽链；PsWD40-2序列包含一个1 032 bp的ORF，编码一个343 aa的肽链。牡丹PsWD40-1和PsWD40-2基因推测所编码蛋白序列均具有典型的WD40结构域。PsWD40-1蛋白序列与葡萄VvWDR2相似性为96%，PsWD40-2蛋白序列与葡萄VvWDR1相似性为87%。系统进化树分析发现，PsWD40-1与葡萄VvWDR2、拟南芥AtAN11、陆地棉GhTTG2等序列聚在MP1分支，而PsWD40-2与葡萄VvWDR1、矮牵牛PhAN11、紫苏PfWD40等序列聚在另一分支TTG1/PAC1分支。

关键词: 牡丹, 花青素苷合成, 调节基因, WD40蛋白

Abstract: Two Unigene sequences that share high homology with WD40 protein involved in plant anthocyanin biosynthesis were obtained from previous-constructed tree peony（Paeonia suffruticosa ‘Luoyang Hong’）petal transcriptome database and named as PsWD40-1 and PsWD40-2. Sequences of open reading frame（ORF）in PsWD40-1 and PsWD40-2 were amplified with designed specific primers using RT-PCR technology and sequenced. Results showed that PsWD40-1 contains a 1 035 bp ORF encoding 344 amino acid residues, and PsWD40-2 contains a 1 032 bp ORF encoding 343 amino acid residues. The predicted protein sequences of PsWD40-1 and PsWD40-2 genes both contain the typical WD40 structural domain. PsWD40-1 shares high similarity（96%）with VvWDR2 of Vitis vinifera, and PsWD40-2 shares 87% similarity with VvWDR1 of Vitis vinifera. Phylogenetic tree analysis showed that PsWD40-1 is grouped into the MP1 clade with VvWDR2 of Vitis vinifera, AtAN11 of Arabidopsis thaliana, and GhTTG2 of Gossypium hirsutum, while PsWD40-2 is grouped into the other clade, TTG1/PAC1 clade, with VvWDR1 of Vitis vinifera, PhAN11 of Petunia hybrida, and PfWD40 of Perilla frutescens.

张超, 高树林, 杜丹妮, 吴凡, 董丽. 牡丹WD40类转录因子基因PsWD40-1和PsWD40-2的分离与序列分析[J]. 生物技术通报, 2014, 0(2): 85-90.

Zhang Chao, Gao Shulin, Du Danni, Wu Fan, Dong Li. Isolation and Sequence Analysis of the Paeonia suffruticosa WD40 Transcription Factor Genes PsWD40-1 and PsWD40-2[J]. Biotechnology Bulletin, 2014, 0(2): 85-90.

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