《植物生理学报》 2011, 47(9): 885-892

牡丹二氢黄酮醇4-还原酶基因PsDFR1的克隆及表达分析

周琳, 王雁*, 任磊, 彭镇华

中国林业科学研究院林业研究所, 国家林业局林木培育重点实验室, 北京100091

通信作者：王雁；Tel: 2011-06-24

摘　要：

二氢黄酮醇4-还原酶(DFR)是花色素苷合成途径中的关键酶, 在植物花色的形成过程中起重要作用。本研究以牡丹 品种‘彩绘’为试材, 采用RT-PCR和RACE方法从花瓣中获得了一个牡丹DFR基因cDNA全长, 命名为PsDFR1, GenBank登录 号为HQ283448。序列分析结果表明, PsDFR1全长1 242 bp, 包含一个长为1 095 bp的开放阅读框, 编码364个氨基酸。生物 信息学分析显示该基因编码的蛋白具有典型的DFR蛋白功能结构域, 包括多个保守的短链脱氢/还原酶家族的特征位点, 属 于NADB_Rossmann超家族。氨基酸序列比对和系统进化树分析表明, PsDFR1与葡萄、棉花、毛果杨、梨等植物的DFR 相似性都在75%以上。实时荧光定量PCR分析表明, PsDFR1在花色素大量积累的花瓣中表达量最高, 其次是萼片和雄蕊, 再次是叶片, 在心皮中表达量最低。

关键词：牡丹; 二氢黄酮醇4-还原酶基因; 克隆; 表达

收稿：2011-06-24   修定：2011-07-14

资助：国家“863”项目(2006AA100109)和国家林业局“948”项目(2006-4-C07)。

Cloning and Expression Analysis of Dihydroflavonol 4-Reductase Gene PsDFR1 from Tree Peony (Paeonia suffruticosa Andr.)

ZHOU Lin, WANG Yan*, REN Lei, PENG Zhen-Hua

Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Corresponding author: WANG Yan; Tel: 2011-06-24

Abstract:

Dihydroflavonol 4-reductase (DFR) is a key enzyme in the anthocyanin biosynthesis pathway, and plays a critical role in flower pigmentation. In this study, a full-length cDNA sequence of DFR gene was obtained from petals of tree peony (Paeonia suffruticosa ‘Cai Hui’) using RT-PCR and RACE, and named PsDFR1 (GenBank accession No. HQ283448). Sequence analysis indicated that PsDFR1 is 1 242 bp in full length and has an open reading frame of 1 095 bp encoding a deduced polypeptide of 364 amino acids. Bioinformatic analysis showed that PsDFR1 had the typical functional domains of DFR protein, containing most conserved characteristic sites in short-chain dehydrogenase/reductase and belonging to the NADB_Rossmann superfamily. Sequence alignment and phylogenetic analysis revealed that PsDFR1 shared more than 75% homology with DFR from grape (Vitis vinifera), cotton (Gossypium hirsutum), Populus trichocarpa and pear (Pyrus communis). Quantitative RT-PCR analysis indicated that PsDFR1 showed the highest transcript abundance in anthocyanin-pigmented petals, moderate levels in sepals and stamens, low level in leaves and the lowest level in carpels.

Key words: tree peony (Paeonia suffruticosa); dihydroflavonol 4-reductase gene; cloning; expression