《植物生理学报》 2017, 53(9): 1772-1780

甘蓝型油菜甘油三磷酸脱氢酶(BnaGPDH)基因的克隆与表达

刘少锋, 刑蔓, 汪慧慧, 邬克彬, 官春云, 肖钢, 熊兴华^*

湖南农业大学农学院, 作物基因工程湖南省重点实验室, 长沙410128

通信作者：熊兴华；E-mail: ndxiongene@yahoo.com

摘　要：

甘油三磷酸脱氢酶(GPDH)是三酰甘油(TAG)合成的关键酶。为了阐明甘蓝型油菜(Brassica napus) GPDH (BnaGPDH)的表达特性, 本研究采用反转录PCR (RT-PCR)方法, 获得BnaGPDH基因的3条全长编码序列(CDS), 分别命名为BnaGPDH1、BnaGPDH2、BnaGPDH3, 长度分别为1 395、1 395、1 389 bp。生物信息学分析表明, BnaGPDHs具有GPDH基因家族的特征, BnaGPDHs与其他植物GPDHs有高度同源性。时空表达分析表明, BnaGPDHs是组成型表达基因, 在花中的表达量最高, 叶中的表达量最低。BnaGPDHs在幼嫩种子发育的第5周表达量普遍提高, 在果荚发育到第5周表达量明显降低。逆境分析表明, BnaGPDHs响应多种逆境胁迫, 可能与植物抗逆机制有关。

关键词：GPDH; 甘蓝型油菜; 基因克隆; 表达分析

收稿：2017-05-15   修定：2017-07-28

资助：国家重点基础研究发展计划(973计划)项目(2015CB-150200)、国家重点研发计划项目(2017YFD0101703)、湖南省科技计划项目(2014FJ 1006-3)和湖南省自然科学基金(2015JJ2080)。

Molecular cloning and expression profile of BnaGPDH gene in Brassica napus

LIU Shao-Feng, XING Man, WANG Hui-Hui, WU Ke-Bin, GUAN Chun-Yun, XIAO Gang, XIONG Xing-Hua^*

Crop Gene Engineering Key Laboratory of Hunan Province, College of Agriculture, Hunan Agricultural University, Changsha 410128, China

Corresponding author: XIONG Xing-Hua; E-mail: ndxiongene@yahoo.com

Abstract:

Glycerol-3-phosphate dehydrogenase (GPDH) is a key enzyme in triacylglycerol (TAG) biosynthesis. In order to elucidate the expression feature of BnaGPDHs in Brassica napus, three GPDH full-length coding sequences (1 395, 1 395 and 1 389 bp) were obtained by reverse transcription PCR, and designated as BnaGPDH1, BnaGPDH2 and BnaGPDH3, respectively. Bioinformatic analysis shows that they have the characteristics of the GPDH family, and amino acid sequences of BnaGPDHs are highly homologous to GPDHs in other plants. Spatio-temporal expression results show that BnaGPDHs are genes of constitutive expression, with the highest expression in flower and the lowest expression in leaf. BnaGPDH expression levels were generally increased in developing seed at 35 days after flowering, at the same time they were significantly decreased in developing capsule. BnaGPDHs responded to abiotic stresses, indicating they might be concerned with the mechanism of plant stress resistance.

Key words: GPDH; Brassica napus; gene cloning; expression analysis