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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (9): 55-59.

所属专题： 生物技术； 园艺； 烟草种植与生产

• 生物技术科学 • 上一篇    下一篇

盐角草甜菜碱合成相关基因的共表达提高转基因烟草的耐盐性

胡艳梅,苏乔,祖勇,刘纪文   

  • 收稿日期:2009-12-08 修回日期:2009-12-23 出版日期:2010-05-05 发布日期:2010-05-05
  • 基金资助:

    辽宁省科技攻关“农业生物技术”

Co-expression of genes related to glycine betaine biosynthesis from Salicornia europaea increase salt tolerance of transgenic tobacco

Hu Yanmei, Su Qiao, Zu Yong, Liu Jiwen   

  • Received:2009-12-08 Revised:2009-12-23 Online:2010-05-05 Published:2010-05-05

PDF (PC)

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摘要：

植物在逆境下能合成甜菜碱，其中磷酸乙醇胺N-甲基转移酶（PEAMT）和胆碱单加氧酶（CMO）是甜菜碱合成过程中的关键酶。本文将盐角草SePEAMT基因、SeCMO基因以及烟草的核基质结合区序列（Mar）利用Cre/loxP重组系统构建到同一表达载体上，得到植物表达载体pYLTAC747N-Mar-SePEAMT-SeCMO-Mar。该载体用电击转化法转入农杆菌EHA105中，通过农杆菌介导法转化烟草，经PCR检测确定转基因植株。在含250mM NaCl的MS培养基上35d后，转基因植株生根而野生型植株不能生根；转基因烟草植株甜菜碱积累量显著高于野生型植株，约是野生型植株的5.6~7.7倍；转基因植株与野生型相比，相对电导率显著降低，叶绿素含量明显升高。以上结果表明共表达SePEAMT 和SeCMO能有效提高烟草甜菜碱表达量从而提高烟草的耐盐性。

关键词: 新农村, 新农村, 跨越式发展, 后发

Abstract:

In plants, betaine is synthesized upon abiotic stress, in which phosphoethanolamine N-methyltransferase（PEAMT）and choline monooxygenase (CMO) are key enzymes. A plant expression vector pYLTAC747N-Mar-SePEAMT-SeCOM-Mar, which includes SePEAMT gene and SeCMO gene cloned from Salicornia europaea, and the matrix attachment region (Mar) isolated from tobacco, was obtained based on Cre/LoxP recombination system. The resultant plasmid was transferred into Agrobacterium tumefaciens EHA105 by the electroporation method, then transferred into tobacco (Nictiana tabacum L. cv. 89) via Agrobacterium-mediated method. PCR analysis showed that genes were transformed successfully into the transgenic plants. In the condition of NaCl stress for 35d, the transgenic plants could root while the wild-type plants could not. The glycine betaine content of transgenic plants was significantly higher than wild type plants, and the glycine betaine content of transgenic plants was about 5.6~7.7 fold of wild plants. Transgenic plants exhibit significantly lower relative conductivity and higher content of chlorophyll. The data suggest that co-expression of SePEAMT and SeCMO might increase the glycine betaine content and improve the salinity tolerance of engineered plants.