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

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

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。

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 RTPCR 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.