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通过农杆菌介导的转化高效生产转基因甜瓜。

Efficient production of transgenic melon via Agrobacterium-mediated transformation.

作者信息

Bezirganoglu I, Hwang S Y, Shaw J F, Fang T J

机构信息

Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan, China.

Department of Entomology, National Chung Hsing University, Taichung, Taiwan, China.

出版信息

Genet Mol Res. 2014 Apr 25;13(2):3218-27. doi: 10.4238/2014.April.25.7.

Abstract

Oriental melon (Cucumis melo L. var. makuwa) is an important fruit for human consumption. However, this plant species is one of the most recalcitrant to genetic transformation. The lack of an efficient in vitro system limits the development of a reproducible genetic transformation protocol for Oriental melon. In this study, an efficient transgenic production method for Agrobacterium-mediated transformation using cotyledon explants of Oriental melon was developed. Cotyledon explants were pre-cultivated for two days in the dark, and the optimal conditions for transformation of melon were determined to be a bacteria concentration of OD600 0.6, inoculation for 30 min, and two days of co-cultivation. Transgenic melon plants were produced from kanamycin-resistant shoots. A total of 11 independent transgenic plants were regenerated with a transformation efficiency of 0.8% of the inoculated explants. The transgenic plants were phenotypically normal and fully fertile, which might be a consequence of the co-cultivation time.

摘要

东方甜瓜(Cucumis melo L. var. makuwa)是一种供人类食用的重要水果。然而,该植物物种是最难进行遗传转化的物种之一。缺乏高效的体外系统限制了东方甜瓜可重复遗传转化方案的开发。在本研究中,开发了一种利用东方甜瓜子叶外植体进行农杆菌介导转化的高效转基因生产方法。子叶外植体在黑暗中预培养两天,甜瓜转化的最佳条件确定为细菌浓度OD600为0.6、接种30分钟以及共培养两天。从卡那霉素抗性芽中获得了转基因甜瓜植株。共再生出11株独立的转基因植株,转化效率为接种外植体的0.8%。转基因植株表型正常且完全可育,这可能是共培养时间的结果。

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