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作物学报 ›› 2013, Vol. 39 ›› Issue (09): 1701-1709.doi: 10.3724/SP.J.1006.2013.01701

• 研究简报 • 上一篇    

大豆钾转运体基因GmKT12的克隆和信息学分析

朱晓玲1,2,陈海峰1,王程1,3,郝青南1,2,陈李淼1,郭丹丹1,2,伍宝朵1,陈水莲1,沙爱华1,周蓉1,*,周新安1,*   

  1. 1中国农业科学院油料作物研究所 / 农业部油料作物生物学重点开放实验室，湖北武汉 430062；2中国农业科学院研究生院，北京 100081；3南京农业大学 / 国家大豆改良中心 /作物遗传与种质创新国家重点实验室，江苏南京 210095
  • 收稿日期:2012-10-15 修回日期:2013-04-22 出版日期:2013-09-12 网络出版日期:2013-07-09
  • 通讯作者: 周蓉, E-mail: zhourong@oilcrops.cn, Tel: 027-86735887; 周新安, E-mail: zhouxa@oilcrops.cn, Tel: 027-86711563
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2011ZX08004-005, 2009ZX08009-133B), 国家自然科学基因项目(30871554, 30900906), 中国农业科学院公益性院所科研基金(1610172011006)资助。

Molecular Cloning and Bioinformatics Analysis of K+ Transporter Gene (GmKT12) from Soybean (Glycine max [L.] Merri)

ZHU Xiao-Ling1,2,CHEN Hai-Feng1,WANG Cheng1,3,HAO Qing-Nan1,2,CHEN Li-Miao1,GUO Dan-Dan1,2,WU Bao-Duo1,CHEN Shui-Lian1,SHA Ai-Hua1,ZHOU Rong1,*,ZHOU Xin-An1,*   

  1. 1 Oil Crops Research Institute of Chinese Academy of Agriculture Sciences / Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Wuhan 430062, China; 2Graduate School of Chinese Academy of Agriculture Sciences, Beijing 100081, China; 3 Nanjing Agricultural University / National Center for Soybean Improvement/ National Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China
  • Received:2012-10-15 Revised:2013-04-22 Published:2013-09-12 Published online:2013-07-09
  • Contact: 周蓉, E-mail: zhourong@oilcrops.cn, Tel: 027-86735887; 周新安, E-mail: zhouxa@oilcrops.cn, Tel: 027-86711563

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摘要：

以钾高效和钾敏感型大豆品系为试验材料，设置低钾胁迫试验，在8个时间段取样提取RNA，利用Real time-PCR检测GmKT12基因的表达量，结果显示GmKT12基因在不同品系地上部和地下部表达水平有显著差异，其原因来自GmKT12基因的氨基酸序列和蛋白质结构的变异。从2个品系中分别克隆目的基因并对基因序列进行同源性及生物信息学分析表明，与GmKT12基因相似性在30%以上的同源基因有56个，GmKT12在进化树中的位置与Glyma18g18822最近；GmKT12编码蛋白为可溶性跨膜蛋白，具有多个磷酸化位点，该基因与信号转导有关，对大豆获取及转运钾离子可能起着关键作用。

关键词: 大豆, 钾转运体基因, 克隆, 生物信息学分析, Real-time PCR, 序列测定, 同源性分析, 系统发育分析

Abstract:

GmKT12 predicted as a potassium transporter is of great importance in absorbing nutrients, signal transduction in plant growth. At present, there are few reports about the gene. In this paper, one low K tolerant soybean line You 06-71 and one low K intolerant line Hengchun 04-11 were weed as plant materials. In order to research the expression level of GmKT12 in low potassium stress conditions, RNA was extracted and tested by Real-time PCR in eight periods. The results showed that there were highly significant differences between two lines due to their was diversity of amino acid sequence and protein structure. Homology and Bioinformatic analysis after cloning the target sequence from the two lines showed that comparing with GmKT12,there were 56 homologous genes with more than 30 percent similarity, GmKT12 andGlyma18g18822 were nearest in the phylogenetic tree. The protein encoded by GmKT12 was soluble transmembrane protein which contained 11–12 membrane structural domains, and possessed multiple phosphorylation sites. These indicated that the protein might take part in plant signal transduction and regulate potassium ions transportation into or out of cells. In conclusion, GmKT12 might play a pivotal role in potassium absorption in soybean, on which the research is important.

Key words: Soybean [Glycine max L. Merri], Potassium transporter, Cloning, Bioinformatics analysis, Real-time PCR, Sequencing, Homologous analysis, Phylogenetic analysi

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