| 小麦蛋白磷酸酶2A调节亚基基因TaBβ-1的克隆及其在非生物胁迫下的表达特性 |
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| 引用本文： | 刘诗航,王彩香,毛新国,刘惠民,李昂,景蕊莲.小麦蛋白磷酸酶2A调节亚基基因TaBβ-1的克隆及其在非生物胁迫下的表达特性[J].中国农业科学,2010,43(11):2197-2208. |
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| 作者姓名： | 刘诗航 王彩香 毛新国 刘惠民 李昂 景蕊莲 |
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| 作者单位： | （山西大学生物工程学院） |
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| 基金项目： | 北京市优秀博士学位论文指导教师项目，国家转基因重大专项课题 |
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| 摘 要： | 【目的】克隆小麦蛋白磷酸酶2A(PP2A)调节亚基(PR55)基因TaBβ-1,分析其在非生物胁迫下的表达特性,为小麦抗逆育种提供候选基因。【方法】以小麦品种旱选10号为材料,通过电子克隆和RT-PCR获得TaBβ-1的全长cDNA序列,采用生物信息学软件分析TaBβ-1及其编码蛋白TaBβ-1的序列特征,预测其功能,利用实时荧光定量PCR(real-time quantitative PCR)技术分析该基因在小麦孕穗期不同组织中、不同生育时期新叶中的表达情况,以及在PEG、NaCl、低温及外源激素ABA等非生物胁迫下的表达模式,检测不同水分条件下成株期转基因拟南芥的叶片细胞膜稳定性。【结果】获得TaBβ-1的全长cDNA序列1931bp,其开放阅读框(ORF)为1539bp。该基因编码512个氨基酸,预测TaBβ-1蛋白分子量为57.1kD,等电点为5.87,含有PP2A调节亚基的1个CDC55保守结构域、1个alpha/beta结构域、2个PR55保守结构域和6个WD重复子。TaBβ-1在小麦孕穗期的根、穗、叶中均有表达,表达量依次为根穗叶;在不同生育时期的新叶中,苗期叶片的表达量最高。TaBβ-1的表达明显受PEG、NaCl、低温以及外源ABA的诱导。【结论】小麦蛋白磷酸酶2A调节亚基家族基因TaBβ-1在小麦苗期叶片中的表达量明显高于根、穗以及其它时期的叶片;TaBβ-1参与对高渗、高盐、低温等多种胁迫以及ABA处理的应答反应,但表达模式不同;在水分胁迫条件下,转基因拟南芥比野生型具有较高的细胞膜稳定性。
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| 关 键 词： | 小麦 TaBβ-1 克隆 表达特性 非生物胁迫 |
| 收稿时间： | 2009-12-28; |
Cloning of Protein Phosphatase 2A Regulatory Subunit Gene TaBβ-1 and Its Expression Patterns Under Abiotic Stresses in Wheat |
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| LIU Shi-hang,WANG Cai-xiang,MAO Xin-guo,LIU Hui-min,Li Ang,JING Rui-lian.Cloning of Protein Phosphatase 2A Regulatory Subunit Gene TaBβ-1 and Its Expression Patterns Under Abiotic Stresses in Wheat[J].Scientia Agricultura Sinica,2010,43(11):2197-2208. |
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| Authors: | LIU Shi-hang WANG Cai-xiang MAO Xin-guo LIU Hui-min Li Ang JING Rui-lian |
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| Affiliation: | （College of Bioengineering, Shanxi University）
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| Abstract: | 【Objective】 TaBβ-1, a gene encoding protein phosphatase 2A (PP2A) regulatory subunit (PR55) was cloned and its expression patterns in wheat under abiotic stresses were dissected for the purpose of providing a candidate gene for wheat breeding in response to various abiotic stresses. 【Method】 A wheat (Triticum aestivum L.) cultivar Hanxuan 10 was used as the plant material to clone the full-length cDNA sequence of TaBβ-1 by in silico cloning and RT-PCR. Suitable bioinformatics softwares were adopted to assay the structure of cDNA sequence, and the function of deduced TaBβ-1. The real-time quantitative PCR (qRT-PCR) method was employed to determine the expression patterns of TaBβ-1 in specific tissues and under abiotic stresses in wheat. 【Result】 A full-length cDNA sequence encoding regulatory subunit of PP2A was cloned from wheat and designated by the name of TaBβ-1, which is a 1 931 bp sequence with an 1 539 bp open reading frame. The putative amino acid sequence of TaBβ-1 contains 512 amino acids with a molecular mass of 57.1 kD and pI value of 5.87, which contains a CDC55 conserved domain, an alpha/beta domain, two PR55 conserved sites and six WD-repeats of PP2A subunit B. TaBβ-1 was detected in all tissues, including root, leaf and young-spike of wheat. The expression levels of TaBβ-1 at booting stage were root > spike > leaf. The highest expression level was identified from the seedling leaf among young-leaves at jointing, booting and flag-leaf expansion stages. TaBβ-1 was up-regulated by PEG, NaCl, cold and ABA treatments. Leaf cell membrane stabilities of transgenic Arabidopsis over-expressing TaBβ-1 were detected under well-watered and drought stress conditions. 【Conclusion】 TaBβ-1 is a member of PP2A regulatory subunit family. The expression level of TaBβ-1 in seedling leaf is much higher than that in the root and spike at booting stage, and in leaves at different stages. TaBβ-1 responds to multi-abiotic stresses but shows varied expression patterns, implying that TaBβ-1 involves in different signal pathways responding to abiotic stresses. Transgenic TaBβ-1 Arabidopsis lines show significantly higher cell membrane stabilities under drought stress condition than the wild type. |
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| Keywords: | wheat TaBβ-1 cloning expression abiotic stress |
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