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作物学报 ›› 2009, Vol. 35 ›› Issue (11): 2029-2036.doi: 10.3724/SP.J.1006.2009.02029

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

转录因子基因TaWRKY72b-1的克隆,表达及在烟草中表达对植株磷效率的影响

苗鸿鹰1,赵金峰1,李小娟2,孙昭华2,路文静2,谷俊涛2,郭程瑾1,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:2008-12-22 修回日期:2009-04-25 出版日期:2009-11-12 网络出版日期:2009-08-07
  • 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(2007CB1162090)项目和河北省重点基础研究项目(08965525D)资助。

Cloning and Expression of Wheat Transcription Factor Gene TaWRKY72b-1 and Its Effect on Phosphorus Use Efficiency in Transgenic Tobacco Plants

MIAO Hong-Ying1,ZHAO Jin-Feng1,LI Xiao-Juan2,SUN Zhao-Hua2,LU Wen-Jing2,GU Jun-Tao2,GUO Cheng-Jin1,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:2008-12-22 Revised:2009-04-25 Published:2009-11-12 Published online:2009-08-07
  • Contact: XIAO Kai, E-mail: xiaokai@hebau.edu.cn

摘要:

在富集低磷胁迫特异表达基因的小麦根系cDNA差减杂交文库中,鉴定了1个与拟南芥WRKY75同源的小麦WRKY型转录因子基因表达序列标签(EST)。依据该EST序列高度同源的小麦WRKY72b序列,克隆了对应基因TaWRKY72b-1TaWRKY72b-1WRKY72bcDNA序列上有2个碱基的差异,但编码氨基酸没有改变。TaWRKY72b-1开放阅读框为621 bp,编码206个氨基酸残基,氨基酸组成上含有保守的WRKY基序和C2H2基序。系统进化分析表明,TaWRKY72b-1与小麦WRKY72a和大麦WRKY12可能来自相同的祖先。与对照供磷水平(2 mmol L-1 P)相比,低磷处理使根叶中TaWRKY72b-1的转录本数量均明显增多。表明TaWRKY72b-1对低磷胁迫逆境产生了明显的应答作用。TaWRKY72b-1在烟草中表达表明,低磷胁迫条件下,高表达TaWRKY72b-1的烟草植株干重、单株磷累积量和磷利用效率均较对照明显增加。因此,TaWRKY72b-1基因在改善低磷胁迫下作物的磷效率中可能具有较重要的应用价值。

关键词: 小麦(Tritium aestivum L.), WRKY转录因子, 基因克隆, 表达, 功能, 磷胁迫

Abstract:

Phosphorus is one of the indispensable elements in plant growth and development, which is the main component for ATP, nucleic acid, and lecithin. Since phosphorus usually exists in the hard-absorption compounds in soil to cause plant suffering from phosphorus deficiency during growing stage. So plant morphological and physiologically develops the adaptation to low phosphorus stress. Transcription regulation plays an important role in responding to deficient-P cue in plants. Several transcription factors mediating the deficient-P signal transduction have been reported in Arabidopsis and rice. But no similar studieshave been conducted in wheat by now. In this study, an expressed sequence tag (EST) homologous to Arabidopsis WRKY75 was identified based on sequencing of clones from a subtractive root cDNA library, in which the differential expressed genes responding to low-P were enriched. The EST gene with high similarity to wheat WRKY72b (GenBank accession No. EF368383), was cloned and referred to TaWRKY72b-1. TaWRKY72b-1 had two base differences with WRKY72b at the cDNA sequence, with an open reading frame (ORF) of 621 bp and encoding a polypeptide of 206 amino acids. TaWRKY72b-1 contained one of conserved WRKY motif and one of C2H2 zinc finger motif. Phylogenetic tree analysis indicated that TaWRKY72b-1, wheat WRKY72a and barley WRKY12  werepossibly derived from one ancestor. Compared with those in sufficient-P (2 mmol L-1 P) condition, the transcripts of TaWRKY72b-1 in roots and leaves under the deficient-P (20 µmol L-1 P) condition were all dramatically increased, suggesting that TaWRKY72b-1 gene was involved in the response to low P stress in the plants. Under deficient-P condition, the expression of TaWRKY72b-1 in transgenic tobacco plants obviously increased plant dry weights, plant accumulative phosphorus amount, and phosphorus utilization efficiency compared with that in CK (empty vector transformed plants). Therefore, the TaWRKY72b-1 gene has possibly a potential use in improving the crop phosphorus use efficiency under low-Pi stress condition.

Key words: Wheat(Triticum aestivum L.), WRKY transcription factor, Gene cloning, Expression, Function, Low phosphorus stress

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