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作物学报 ›› 2009, Vol. 35 ›› Issue (9): 1749-1754.doi: 10.3724/SP.J.1006.2009.01749

• 研究简报 • 上一篇    下一篇

小麦应答低磷的钙依赖蛋白激酶基因TaCPK1A和TaCPK10的克隆和表达

路文静2，李瑞娟2，李小娟2，郭程瑾1，谷俊涛2，肖凯1,*   

  1. 1河北农业大学农学院，河北保定071001；2河北农业大学生命科学学院；河北保定071001
  • 收稿日期:2009-02-27 修回日期:2009-04-30 出版日期:2009-09-12 网络出版日期:2009-07-04
  • 通讯作者: 肖凯, E-mail: xiaokai@hebau.edu.cn
  • 基金资助:

    本研究由国家重点基础研究发展计划（973计划）前期项目（2007CB116209），河北省重点基础研究项目（08965525D），河北省自然科学基金（C2007000476）资助。

Cloning and Expression of Calcium-Dependent Protein Kinase Genesis TaCPK1A and TaCPK10 in Response to Deficient-Pi in Wheat

LU Wen-Jing2,LI Rui-Juan2,LI Xiao-Juan2,GUO Cheng-Jin1,GU Jun-Tao2,XIAO Kai1*   

  1. 1 College of Agronomy, Agricultural University of Hebei, Baoding 071001, China; 2 College of Life Science, Agricultural University of Hebei, Baoding 071001, China
  • Received:2009-02-27 Revised:2009-04-30 Published:2009-09-12 Published online:2009-07-04
  • Contact: XIAO Kai, E-mail: xiaokai@hebau.edu.cn

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3509

被引次数

10

摘要：

采用构建富集磷胁迫特异表达基因cDNA差减文库、序列分析和cDNA-AFLP技术，鉴定了2个应答低磷胁迫的钙依赖蛋白激酶(CDPK)基因的表达序列标签。克隆、测序和比对结果表明，上述基因分别为TaCPK1A和TaCPK10。其cDNA长度分别为2 129 bp和1 696 bp，开放阅读框分别为1 599 bp和1 281 bp，分别编码532和426个氨基酸；具有CDPK的典型结构特征。系统进化分析表明，上述基因的核苷酸序列同源性低，分别来自不同的祖先。在对低磷胁迫的响应上，TaCPK1A在磷胁迫1~24 h范围内根系内的表达水平不断增强，叶内表达水平在1 h内明显被诱导，以后保持稳定；TaCPK10在相应磷胁迫时间范围根叶内的表达水平均呈低—高—低变化，在磷胁迫1 h的表达被诱导，以后又逐渐降至胁迫前水平。TaCPK1A和TaCPK10对氮、钾胁迫没有应答响应。结果表明，CDPK在介导小麦低磷胁迫的信号转导中具有重要作用，小麦中存在两种或多种CDPK介导的磷酸化过程参与低磷信号的转导。

关键词: 小麦（Triticum aestivum L.）, 钙依赖蛋白激酶, 克隆, 低磷胁迫, 基因表达

Abstract:

Two expressed sequence tags (ESTs) of calcium-dependent protein kinase (CDPK) genes responding to deficient-Pi were identified from wheat (Triticum aestivum L.) cultivar Shixin 828 by subtractive suppression cDNA library and cDNA-AFLP approaches. The genes, TaCPK1A and TaCPK10, contained the conserved domains of plant CDPK, were 2 129 bp and 1 696 bp in full length open reading frames of 1599 bp and 1 281 bp, encoding 532 and 426 amino acids,respectively. Phylogenetic analysis implied different ancestors of TaCPK1A and TaCPK10 because of their low identity of sequence. Under low-Pi condition, the expression level of TaCPK1A in roots was strongly inducible and reached the highest at 24 h after treatment, but that in leaves was induced in 1 h of treatment and maintained stably afterwards. The expression of TaCPK10 in roots and leaves both showed a pattern of low–high–low with the peak at 1 h of treatment,and then decreased to the level before treatment. No responses of TaCPK1A and TaCPK10 were observed to low-N and low-K stresses. Therefore, it is suggested that CDPK playsan important role in mediating phosphate-starvation signal in wheat. There are at least two phosphorylation reaction pathways for transduction of low-Pi signal, in which CDPKs are involved.

Key words: Wheat(Triticum aestivum L.), Calcium-dependent protein kinase, Cloning, Low-Pi stress, Gene expression

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