中国农业科技导报 ›› 2017, Vol. 19 ›› Issue (10): 21-27.DOI: 10.13304/j.nykjdb.2016.657

• 生物技术 生命科学 • 上一篇    下一篇

陆地棉钾吸收相关基因GhHAK1的克隆与功能分析

王昭玉1,吕婷婷1,李爱芹2,王頔1,张子玉1,闫瑞娴1, 陈佳雪1,董惠心1,唐宁蔓3,刘建凤1*   

  1. 1.河北大学生命科学学院, 河北 保定 071002; 2.河北省承德市兴隆县农牧局, 河北 兴隆 0673000; 3.河北大学经济学院, 河北 保定 071002
  • 收稿日期:2016-10-26 出版日期:2017-10-15 发布日期:2016-11-21
  • 通讯作者: 通信作者:刘建凤,副教授,博士,研究方向为植物逆境胁迫机制。E-mail:jianfengliu@hbu.edu.cn
  • 作者简介:王昭玉,硕士研究生,研究方向为植物分子生物学。E-mail:1023988344@qq.com。
  • 基金资助:
    河北省自然科学基金项目(C2015201227);河北大学人才引进项目(2014-277)资助。

Cloning and Fuctional Characterization of a Potassium Absorption Related Gene GhHAK1 from Cotton (Gossypium hirsutum L.)

WANG Zhaoyu1, LV Tingting1, LI Aiqin2, WANG Di1, ZHANG Ziyu1, YAN Ruixian1, CHEN Jiaxue1, DONG Huixin1, TANG Ningman2, LIU Jianfeng1*   

  1. 1.College of Life Science, Hebei University, Hebei Baoding 071002; 2.Xinglong County Agriculture and Animal Husbandry Bureau of Chengde city in Hebei Province, Hebei Chengde 067300; 3.School of Economics, Hebei University, Hebei Baoding 071002, China
  • Received:2016-10-26 Online:2017-10-15 Published:2016-11-21

摘要: 棉花是一种典型的喜钾经济作物,其产量和品质形成与钾含量密切相关。克隆获得棉花高亲和性钾转运体基因GhHAK1框,系统进化树表明该GhHAK1基因属于第Ⅰ亚族,此分支被认为介导高亲和性K+。膜结构预测显示该基因编码的蛋白跨膜螺旋区域(TMS)有 12个,且在第2和第3 TMS之间有一个较长的胞质质环。洋葱表皮亚细胞定位表明,该基因编码蛋白定位在细胞质膜上;继而对部分转GhHAK1基因拟南芥纯合系进行K+营养试验,发现在低钾(0.05 mmol/L)条件下,野生型拟南芥表现明显的缺钾症状,叶片黄化,植株抽薹明显受抑制,而转基因材料则表现植株叶片生长正常,抽薹未受影响,同时转基因拟南芥叶片中K+含量约为野生型拟南芥的2倍左右,根中K+含量约为1.5倍左右。该结果初步表明GhHAK1基因具有介导植株钾高效吸收的功能,为进一步培育适应土壤钾素匮乏及盐渍化环境下生长的棉花新品种提供重要的基因资源。

关键词: 陆地棉, GhHAK1, 克隆, 低钾胁迫

Abstract: Cotton is an important economic crop typically with preference for potassium. Its yield and quality are closely related to potassium content. In this study, open read frame of GhHAK1 was cloned from the upland cotton. The dendrograms of GhHAK1 showed that it belonged to theⅠsubtribe, which was considered to be an important component of high-affinity system for K+. The trans-membrane region prediction reveals that the protein coded by this gene with 12 trans-membrane domains (TMS), and it had a long cytoplasmic ring between the second and third TMS. The sub-cellular localization experiment of onion epidermis showed that GhHAK1 was located on plasma membrane. And under the condition of low potassium (0.05 mmol/L) condition, the wild-type Arabidopsis showed obvious symptoms of K deficiency, the leaves turned yellow and bolting was obviously inhibited. But the transgenic plants growth was not affected. Besides, the content of K+ in transgenic Arabidopsis leaves was double that of the wild type and the content of K+ in root was about 1.5 times. The results showed that GhHAK1 might possess efficient potassium absorption function and would provide important genetic resources for further breeding new cotton varieties adapted to potassium deficiency soil and saline environment.

Key words: cotton (Gossypium hirsutum L.), GhHAK1, clone, low potassium stress