《植物生理学报》 2017, 53(4): 536-544

牡丹开花调控转录因子基因PsFUL1的克隆与表达分析

位伟强^1,2, 刘伟^1,2, 段亚宾^1,2, 郭丽丽^1,2, 张利霞^1,2, 郭大龙^2,3,*, 侯小改^1,2,∗

¹河南科技大学农学院/牡丹学院, 河南洛阳471003; ²河南省油用牡丹工程技术研究中心, 河南洛阳471003; ³河南科技大学林学院, 河南洛阳471003

通信作者：郭大龙；E-mail: grapeguo@126.com; hxg382@126.com

摘　要：

以‘洛阳红’牡丹为试验材料, 采用RT-PCR方法从其花芽中克隆得到1个开花调控重要转录因子基因FRUITFULL (FUL1)的同源基因, 其cDNA开放阅读框长度为768 bp, 编码255个氨基酸。序列比对和结构域分析表明, 此蛋白具有MADS超家族、K-box超家族的结构域和ARG80多结构域, 且具有MADS-box和K-box的稳定保守基序。NCBI blast分析发现克隆得到的氨基酸序列与葡萄中的FUL类蛋白具有较高同源性, 为69%。因此将其命名为PsFUL1, GenBank登录号为KX621277。系统进化树分析表明, PsFUL1与葡萄编码的VvFUL1亲缘关系最近, 属于MADS基因家族中的AP1/FUL亚家族。qRT-PCR表明, PsFUL1基因在‘洛阳红’牡丹不同组织中表达差异显著, 在花芽和花瓣中的表达量最高, 苞片和叶片次之, 根中最少。在不同品种牡丹(早花‘迎日红’、中花‘洛阳红’、晚花‘花王’)的6个不同开花时期的花瓣中, PsFUL1基因表达量因花期早晚与开花时期不同而差异显著, 表明PsFUL1基因对牡丹开花及花期的早晚有重要调控作用。

关键词：牡丹; PsFUL1; 不同花期; 基因表达量

收稿：2016-10-27   修定：2017-03-02

资助：河南省高校科技创新团队支持计划(14IRTSTHN014)、国家自然科学基金(31370697)和河南省科技创新杰出人才基金(164200510013)。

Molecular cloning and expression analysis of flowering-regulating transcription factor gene PsFUL1 in tree peony (Paeonia suffruticosa)

WEI Wei-Qiang^1,2, LIU Wei^1,2, DUAN Ya-Bin^1,2, GUO Li-Li^1,2, ZHANG Li-Xia^1,2, GUO Da-Long^2,3,*, HOU Xiao-Gai^1,2,*

¹College of Agriculture/Peony, Henan University of Science and Technology, Luoyang, Henan 471003, China; ²Henan Engineering Research Center for Oil Peony, Luoyang, Henan 471003, China; ³College of Forestry, Henan University of Science and Technology, Luoyang, Henan 471003, China

Corresponding author: GUO Da-Long; E-mail: grapeguo@126.com; hxg382@126.com

Abstract:

One important flowering-regulating transcription factor gene FRUITFULL (FUL1) homologous gene was obtained from tree peony (Paeonia suffruticosa ‘Luoyanghong’) by RT-PCR method. Open reading frame (ORF) length of its cDNA was 768 bp, and could encode 255 amino acids. Sequence alignment and structure domain analysis showed that the protein possessed MADS super family, K-box super family structure domains and multi-domains in ARG80. Also, the protein had stable conservative MADS-box and K-box sequences. NCBI blast analysis found that cloned amino acid sequences had high homology at 69% compared with the FUL-like proteins in Vitis vinifera. This gene was therefore designated as PsFUL1 and GenBank accession number was KX621277. Phylogenetic analysis showed that it had close relationship with VvFUL1 in V. vinifera, belonging to subfamily AP1/FUL in MADS gene families. The results of qRT-PCR showed that PsFUL1 gene displayed significant expression difference in different organs of P. suffruticosa ‘Luoyanghong’. High expression levels were observed in the bud and petal, followed by bract, leaf and root. Among the six different flowering stages of different varieties of tree peony (early-flowering ‘Yingrihong’, mid-flowering ‘Luoyanghong’, late-flowering ‘Huawang’), PsFUL1 gene expression level had significant difference with different flowering stages and blossoms. The results indicated that PsFUL1 gene had an important regulation effect on the flower formation and blossom in tree peony.

Key words: tree peony (Paeonia suffruticosa); PsFUL1; different flowering stages; gene expression level