《植物生理学报》 2017, 53(9): 1728-1734

蓝莓花芽休眠相关MADS-box基因VcDAM1的克隆与表达分析

刘思, 夏秀英^*, 李波

大连理工大学生命科学与技术学院, 辽宁大连116024

通信作者：夏秀英；E-mail: xx47@dlut.edu.cn

摘　要：

休眠相关MADS-box (DAM)基因在植物芽休眠及解除过程中发挥重要作用。本研究以‘蓝丰’蓝莓(Vaccinium corymbosum)休眠花芽为材料, 利用RT-PCR及RACE技术, 克隆获得VcDAM1基因(GenBank登录号为KF871069)。该基因全长1 376 bp, 包含一个750 bp的开放阅读框, 编码249个氨基酸。序列比对发现, 该基因编码的蛋白质具有典型的MIKCC型MADS-box结构域和K-box结构域, 其氨基酸序列与砂梨DAM同源性较高。实时定量PCR结果表明, 该基因在蓝莓花芽中表达量最高, 且随花芽休眠进程呈现规律变化, 在内休眠期间表达量持续上升, 内休眠解除后表达量迅速下降; 外源脱落酸能使该基因表达上调, 外源赤霉素和单氰胺使该基因表达下调。推测该基因调控蓝莓花芽的休眠。

关键词：蓝莓; 休眠相关MADS-box基因; 休眠; 克隆; 表达分析

收稿：2017-04-17   修定：2017-09-18

资助：教育部留学归国人员启动金项目(第48批)。

Cloning and expression analysis of dormancy-associated MADS-box gene VcDAM1 from blueberry (Vaccinium corymbosum)

LIU Si, XIA Xiu-Ying^*, LI Bo

School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116024, China

Corresponding author: XIA Xiu-Ying; E-mail: xx47@dlut.edu.cn

Abstract:

Dormancy-associated MADS-box (DAM) genes play important roles in regulating bud dormancy induction and release in plants. In this study, a MADS-box gene, named as VcDAM1 (GenBank accession number KF871069) was isolated from the dormant floral buds of blueberry (Vaccinium corymbosum) using RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The gene consists of 1 376 bp which has a 750-bp open reading frame (ORF) coding a protein of 249 amino acids. Multiple sequence comparison revealed that Vc-DAM1 encodes a protein, which is the typical MIKCC-type MADS-box protein with the MADS-box domain and K-box region. Phylogenetic analysis indicated that the amino acid sequence of VcDAM1 had high homology with dormancy-associated MADS-box from Pyrus pyrifolia. qRT-PCR analysis indicated that the gene expression level in flower buds was the highest among all the organs, and with the process of flower bud dormancy, the gene expression level changed regularly. The gene expression level was increased during dormancy, and decreased rapidly after the dormancy was released. Exogenous abscisic acid (ABA) up-regulated the expression of VcDAM1, while exogenous gibberellin (GA) and hydrogen cyanamide (HC) down-regulated the expression of VcDAM1. The results imply that VcDAM1 gene might play an important role in regulating bud dormancy in blueberry.

Key words: Vaccinium corymbosum; dormancy-associated MADS-box gene; dormancy; clone; expression analysis