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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
引用本文:路文静,李瑞娟,李小娟,郭程瑾,谷俊涛,肖凯.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[J].作物学报,2009,35(9):1749-1754.
作者姓名:路文静  李瑞娟  李小娟  郭程瑾  谷俊涛  肖凯
作者单位: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
基金项目:国家重点基础研究发展计JOJ(973计划)前期项目,河北省重点基础研究项目,河北省自然科学基金 
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
收稿时间:2009-02-27

Cloning and Expression of Calcium-Dependent Protein Kinase Genes TaCPK1A and TaCPK10 in Response to Deficient-Pi in Wheat
LU Wen-Jing,LI Rui-Juan,LI Xiao-Juan,GUO Cheng-Jin,GU Jun-Tao,XIAO Kai.Cloning and Expression of Calcium-Dependent Protein Kinase Genes TaCPK1A and TaCPK10 in Response to Deficient-Pi in Wheat[J].Acta Agronomica Sinica,2009,35(9):1749-1754.
Authors:LU Wen-Jing  LI Rui-Juan  LI Xiao-Juan  GUO Cheng-Jin  GU Jun-Tao  XIAO Kai
Institution:1. College of Agronomy, Agricultural University of Hebei, Baoding 071001, China;2. College of Life Science, Agricultural University of Hebei, Baoding 071001, China
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.
Keywords:Wheat(Triticum aestivum L  )  Calcium-dependent protein kinase  Cloning  Low-Pi stress  Gene expression
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