《植物生理学报》 2011, 47(1): 91-96

商陆高亲和性K⁺ 转运体基因PaHAK1 的克隆与表达分析

蔺万煌¹, 马立英¹, 苏益¹, 黄志刚¹, 萧浪涛^1,*, 周清明^2,*

湖南农业大学1 植物激素与生长发育湖南省重点实验室, 2 农学院, 长沙410128

通信作者：萧浪涛；E-mail: langtaoxiao@163.com, zqm8051@hunau.net；Tel: 0731-84635260

摘　要：

为了研究高钾植物商陆高亲和性 K+ 吸收机制, 本试验选用野生商陆为材料, 以拟南芥、冰叶日中花、小麦、大麦和 水稻等多种植物的 HAK 家族基因的保守序列, 设计简并引物, 获得了 981 bp 的 PaHAK1 的基因片段。然后应用 RACE 技 术克隆到PaHAK1 基因序列, 其 cDNA 全长为2 337 bp, 编码771 个氨基酸。该编码蛋白质分子量为86.66 kDa, 等电点为7.54。 蛋白质疏水性和拓扑结构分析显示该氨基酸序列共有12~13个跨膜区。该cDNA序列与冰叶日中花HAK1基因序列相似性 高达 88%。Real-Time PCR 分析表明 PaHAK1 主要在商陆根中表达, 低钾状态可以诱导 PaHAK1 的高表达。

关键词：商陆; 高亲和性K+转运体; 基因克隆; 表达分析

收稿：2010-10-25   修定：2010-11-23

资助：国家自然科学基金项目(30900099)、湖南省自然科学基金项目(09JJ3052)和湖南湖南省教育厅青年基金项目(03B0 11)。

Cloning and Expression Analysis of High-Affinity Potassium Transporter Gene PaHAK1 from Phytolacca acinosa Roxb.

LIN Wan-Huang¹, MA Li-Ying¹, SU Yi¹, HUANG Zhi-Gang¹, XIAO Lang-Tao^1,*, ZHOU Qing-Ming^2,*

¹Hunan Provincial Key Laboratory of Phytohormones and Growth Development, ²College of Agronomy, Hunan Agricultural University, Changsha 410128, China

Corresponding author: XIAO Lang-Tao; E-mail: langtaoxiao@163.com, zqm8051@hunau.net; Tel: 0731-84635260

Abstract:

To study the mechanism of high affinity potassium absorption in some high-K plants, the PaHAK1 cDNA fragment of 981 bp sequences was cloned from wild Phytolacca acinosa Roxb. by polymerase chain reaction with degenerate primers designed according to conserved HAKs cDNA sequences of Arabidopsis thaliana, Mesembryanthemum crystallinum, Triticum aestivum, Hordeum vulgare and Oryza sativa. The fulllength cDNA sequences of PaHAK1 was 2 337 bp which was isolated with rapid amplification of cDNA ends method. The deduced protein was 771 amino acids with molecular weight 86.66 kDa and isoelectric point 7.54. Hydrophobicity plot and topology prediction results indicated the protein had 12–13 putative transmembrane regions. Blastn results showed it was 88% homology to HAK1 of Mesembryanthemum crystallinum. The RealTime PCR results showed the PaHAK1 was mostly expressed in root of the plant and it could be induced in the low potassium condition.

Key words: Phytolacca acinosa Roxb.; high-affinity potassium transporter; gene cloning; expression analysis