《植物生理学报》 2009, 45(12): 1155-1159

二色补血草液泡膜H⁺-ATP 酶C 亚基(LbVHA-C)基因克隆和表达分析

蒋丽丽, 杨传平, 徐晨曦, 马辉, 王玉成*

东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨150040

通信作者：王玉成；E-mail: wangyucheng1029@yahoo.com.cn；Tel: 0451-82190607

摘　要：

从二色补血草 cDNA 文库中分离出一个 V-H+-ATP 酶 C 亚基(LbVHA-C)基因全长 cDNA 序列。该基因全长为 729 bp。 其中, 开放读码框(ORF)为 498 bp, 编码 165 个氨基酸, 预测编码蛋白的分子量为 16.6 kDa, 理论等电点为 8.62。用实时定量 RT-PCR 方法进一步研究二色补血草在 NaCl、NaHCO3 和 NaCO3 胁迫下的不同时间内该基因表达模式的结果表明, NaCl 强烈诱导二色补血草叶组织中 LbVHA-C 基因的表达, 但其根部的表达变化不明显。NaHCO3 胁迫下LbVHA-C 基因在根部 组织的表达受抑制。Na2CO3 胁迫24 h 时的LbVHA-C 基因在根和叶部组织中的表达都强烈受抑制, 随后在根和叶组织的表 达量逐步升高。

关键词：二色补血草; V-H+-ATP 酶 C 亚基基因; 实时定量 RT-PCR; 基因表达

收稿：2009-06-30   修定：2009-11-08

资助：黑龙江省青年科技专项(QC07C56)和教育部科学技术研究重点项目(107037 )。

Cloning and Expression Analysis of LbVHA-C Gene from Limonium bicolor (Bunge) Kuntze

JIANG Li-Li, YANG Chuan-Ping, XU Chen-Xi, MA Hui, WANG Yu-Cheng*

Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, Harbin 150040, China

Corresponding author: WANG Yu-Cheng; E-mail: wangyucheng1029@yahoo.com.cn; Tel: 0451-82190607

Abstract:

A novel vacuolar H⁺-ATPase gene (LbVHA-C) was cloned from a cDNA library of Limonium bicolor. Sequence analysis showed that the LbVHA-C gene was 729 bp in length, including 498 bp of open reading frame (ORF), and encodes a protein of 165 amino acids with a predicted molecular mass of 16.6 kDa and pI of 8.62. The expression of LbVHA-C gene of L. bicolor in response to NaCl, NaHCO₃ and Na₂CO₃ were investigated by real-time RT-PCR. The result showed that the expression of LbVHA-C gene was strongly induced by NaCl in leaves of L. bicolor, but not highly changed in roots. Under NaHCO₃ stress, the expression of LbVHAC gene was inhibited in roots. And the expression of LbVHA-C gene were also strongly inhibited in both roots and leaves under NaCO₃ stress for 24 h, then the expression increased.

Key words: Limonium bicolor; vacuolar H⁺-ATPase gene (LbVHA-C); real time RT-PCR; gene expression