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作物学报 ›› 2016, Vol. 42 ›› Issue (08): 1112-1121.doi: 10.3724/SP.J.1006.2016.01112

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

马铃薯HD-Zip I家族ATHB12基因的克隆及功能鉴定

武亮亮,姚磊,马瑞,朱熙,杨江伟,张宁,司怀军   

  1. 甘肃省作物遗传改良与种质创新重点实验室,甘肃省干旱生境作物学省部共建国家重点实验室培育基地 / 甘肃农业大学生命科学技术学院,甘肃兰州 730070
  • 收稿日期:2016-01-19 修回日期:2016-05-09 出版日期:2016-08-12 网络出版日期:2016-06-02
  • 通讯作者: 张宁, E-mail: ningzh@gsau.edu.cn, Tel: 0931-7631875
  • 基金资助:

    本研究由国家自然科学基金项目(31460370),高等学校博士学科点专项科研基金项目(20126202110007),国家国际科技合作专项(0102014DFG31570)和甘肃省干旱生境作物学省部共建国家重点实验室培育基地开放基金项目(GSCS-2012-02)资助。

Cloning and Functional Identification of the ATHB12 Gene of HD-Zip IFamily in Potato(Solanum tuberosum L.)

WU Liang-Liang,YAO Lei,MA Rui,ZHU Xi,YANG Jiang-Wei,ZHANG Ning*,SI Huai-Jun   

  1. Gansu Key Laboratory of Crop Genetic and Germplasm Enhancement, Gansu Provincial Key Laboratory of Aridland Crop Science /College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2016-01-19 Revised:2016-05-09 Published:2016-08-12 Published online:2016-06-02
  • Contact: 张宁, E-mail: ningzh@gsau.edu.cn, Tel: 0931-7631875
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31460370), Specialized Research Fund for the Doctoral Program of Higher Education of China (20126202110007), International Science & Technology Cooperation Program of China (0102014DFG31570) and Gansu Key Laboratory of Aridland Crop Science of Gansu Agricultural University (GSCS-2012-02).

摘要:

HD-Zip I是一类植物特异转录因子, 在植物应对外界逆境胁迫过程中有重要的调控作用。本研究从马铃薯栽培品种甘农薯2号中克隆了HD-Zip I转录因子ATHB12基因, 其开放阅读框为759 bp, 编码252个氨基酸残基。该基因位于马铃薯1号染色体, 其启动子区含有ABRE、LTRECOREATCOR15和WBOXATNPR1等多种响应非生物胁迫(ABA、低温、脱水及高盐)的顺式作用元件。ATHB12基因在马铃薯根、茎和叶中均有表达, 其根中表达量最高。qRT-PCR分析证实该基因的表达受聚乙二醇(PEG)、NaCl和ABA诱导, 受低温抑制。构建了由组成型表达启动子CaMV 35S驱动的ATHB12基因的过表达载体, 通过农杆菌介导法转化马铃薯获得了转基因植株。干旱处理后, 转基因植株叶片中丙二醛含量显著低于非转基因植株(P<0.05), 而脯氨酸含量显著高于非转基因植株(P<0.05); 转基因植株根鲜重和干重均高于非转基因植株; 说明马铃薯ATHB12基因可能参与了逆境胁迫的响应。

关键词: 马铃薯, HD-Zip, ATHB12基因, 根, 丙二醛, 脯氨酸

Abstract:

HD-Zip I is a class of plant-specific transcription factors, which has an important role in response to adversity stress in plant. A ATHB12 gene of HD-Zip I transcription factors was cloned from potato cultivar Gannongshu2, which contains a 759 bp open reading frame (ORF) encoding a protein of 252 amino acid residues. ATHB12 gene is located on potato chromosome 1, and its promoter region sequence contains cis-acting elements including ABRE, LTRECOREATCOR15, WBOXATNPR1 responsive to abiotic stresses (ABA, temperature, dehydration and salt stress). ATHB12 gene expressed in root, stem and leaf of potato, with the highest expression in the root. qRT-PCR analysis confirmed that the gene was induced by PEG, NaCl and ABA, but repressed by cold treatment. The overexpressed-vector of ATHB12 gene driven by the constitutive promoter CaMV 35S was constructed, and the transgenic plants were obtained using Agrobacterium-mediated transformation system. The malondialdehyde (MDA) content in the transgenic plant leaves was significantly lower (P<0.05), whereas the proline content was significantly higher (P<0.05) than those of non-transgenic control under drought stress. The fresh and dry weight of the transgenic plant root was higher than that of non-transgenic plants. These results showed that ATHB12 gene may be involved in response to stress.

Key words: Potato, HD-Zip, THB12 gene, Root, Malondialdehyde, Proline

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