《植物生理学报》 2013, 49(5): 445-451

地黄内质网型(ER) Ca²⁺-ATPase基因的克隆及表达分析

刘驰¹, 李明杰¹, 王鹏飞¹, 杨艳会³, 王丰青¹, 张重义^1,2, 李春奇¹, 陈新建^1,*

1河南农业大学生命科学学院, 郑州450002; 2福建农林大学中药GAP研究所, 福州350002; 3河南工业大学生物工程学院, 郑 州450001

通信作者：陈新建；E-mail: xinjian@371.net；Tel: 0371-63558722

摘　要：

在分析和筛选我们前期建立的地黄转录组文库、抑制消减杂交文库及地黄相关EST文库基础上, 利用拼接、延伸及 PCR方法, 成功克隆到一个全长2 681 bp的Ca2+-ATPase基因, 含有2 007 bp完整的开放阅读框, 编码568个氨基酸。分析表明,地黄Ca2+-ATPase属于内质网型ATPase, 与大豆和葡萄有较近亲缘关系, Ca2+-ATPase含有Ca2+离子转运区、ATP结合区等保守区。随着生长, 正茬地黄Ca2+-ATPase表达量逐渐升高, 而连作地黄中的Ca2+-ATPase基因在块根伸长期表达突然升高, 之后降低。

关键词：地黄; 连作障碍; 钙信号; Ca2+-ATPase

收稿：2013-01-18   修定：2013-03-23

资助：国家自然科学基金(30973875、81072983和81274022)。

Cloning and Expression Analysis of Endoplasmic Reticulum Type (ER) Ca²⁺-ATPase in Rehmannia glutinosa Libosch

LIU Chi¹, LI Ming-Jie¹, WANG Peng-Fei¹, YANG Yan-Hui³, WANG Feng-Qing¹, ZHANG Zhong-Yi^1,2, LI Chun-Qi¹, CHEN Xin-Jian^1,*

¹School of Life Sciences, Henan Agricultural University, Zhengzhou 450002, China; ²GAP Institute of Chinese Herbal Medicine, Fujian Agriculture and Forestry University, Fuzhou 350002, China; ³School of Biological Engineering, Henan University of Tech-nology, Zhengzhou 450001, China

Corresponding author: CHEN Xin-Jian; E-mail: xinjian@371.net; Tel: 0371-63558722

Abstract:

Based on our previous constructed Rehmannia glutinosa transcriptome, suppression subtractive hybridization (SSH) libraries and EST, by using splicing, extension and PCR methods, we successfully cloned a 2 681 bp full length cDNA sequence of Ca2+-ATPase gene containing an open-reading frame (ORF) with 2 007 bp encoding 568 amino acids. The results indicated that Rehmannia glutinosa Ca2+-ATPase belonged to the endoplasmic reticulum type, had a closer genetic relationship with soybean and grape, and contained a Ca2+ ion transport area and a conservative ATP-binding domain. The expression level of Ca2+-ATPase gradually increased with the grow in first-planting of R. glutionosa, but in continuous cropping of R. glutionosa appeared a higher peak during the root extend period and then decreased.

Key words: Rehmannia glutinosa; continuous cropping obstacle; calicium signal; Ca²⁺-ATPase