牡丹切花ERF转录因子基因的分离与表达分析  

Isolation and Expression Analysis of ERF Transcription Factor Genes in Tree Peony Cut Flowers

作　　者：吴凡[1] 张超[2] 郭加 刘爱青 董丽[1] 

机构地区：[1]北京林业大学园林学院,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,北京100083 [2]浙江农林大学风景园林与建筑学院,浙江临安311300 [3]菏泽市曹州牡丹园,山东菏泽274000

出　　处：《园艺学报》2016年第1期109-120,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31572164);高等学校博士学科点专项科研基金项目(20130014110014)

摘　　要：利用已获得的‘洛阳红’牡丹花瓣转录组数据库,筛选得到3个与植物乙烯响应因子ERF蛋白同源性较高的Unigene序列,利用RACE及RT-PCR技术,设计特异性引物对3个ERF基因最大阅读框进行扩增并测序。生物信息学分析表明PsERF1、PsERF2和PsERF3分别含有长为1 158、747和609 bp的开放阅读框,编码383、248和202个氨基酸残基,且均含有1个典型的AP2结构域。进化分析表明3个牡丹ERF转录因子分别属于ERF亚家族的B2组、B1组和B3组。利用实时定量PCR分析了3个ERF基因在牡丹乙烯敏感型品种‘洛阳红’和乙烯不敏感型品种‘雪映桃花’不同花器官(花瓣、雄蕊、雌蕊和萼片)以及不同发育级别切花花瓣中的表达情况。结果表明,Ps ERF1在2个品种花瓣中的表达量显著高于其他花器官,其表达量在‘洛阳红’切花发育过程中逐渐增加,而在‘雪映桃花’切花发育过程中逐渐降低,推测PsERF1可能对牡丹切花的乙烯响应具有重要的调控作用,PsERF2和PsERF3则可能同时参与了多种信号途径。Three Unigene sequences that share high homology with ERF protein involved in plant ethylene response were obtained from the petal transcriptome database of tree peony（Paeonia suffruticosa ‘Luoyanghong＇）. Sequences of open reading frame in three ERF genes were amplified with designed specific primers using RACE and RT-PCR technology. Bioinformatics analysis indicated that Ps ERF1,Ps ERF2 and Ps ERF3 had a 1 158,747 and 609 bp open reading frame,encoding 383,248 and 202 amino acids,respectively. All the three proteins contained a typical AP2 domain conserved in AP2/ERF family transcription factors. Phylogenetic analysis showed that the three ERF transcription factors belong to B2,B1 and B3 group of ERF subfamily,respectively. An ethylene-sensitive cultivar‘Luoyanghong＇and an ethylene-insensitive cultivar‘Xueying Taohua＇were selected to analyze the expression profiles of three ERF genes in different floral tissues（petals,stamens,pistils or sepals）and in petals of different developmental stages by quantitative real-time PCR. Comparing to other floral tissues,transcript of Ps ERF1 was the most abundant in petals of both cultivars,which increased during development of ‘Luoyanghong＇cut flowers while decreased during that of‘Xueying Taohua＇cut flowers,implying an important role of this gene in ethylene response of tree peony cut flowers. Ps ERF2 and Ps ERF3 might be involved in more than one signal pathway.

关 键 词：牡丹 切花 ERF类转录因子 乙烯敏感性 

分 类 号：S685.11[农业科学—观赏园艺]