A Stable and Efficient Agrobacterium tumefaciens-Mediated Genetic Transformation of the Medicinal Plant Digitalis purpurea L.
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- 作者：Ying Li (1) (3)
Zhenrui Gao (1) (3)
Chunlan Piao (1)
Kaiwen Lu (2)
Zhiping Wang (2)
Min-Long Cui (1) - 关键词：Digitalis purpurea L ; Common medicinal plant ; Mature leaf explants ; High ; efficiency shoot regeneration ; Agrobacterium tumefaciens ; mediated transformation ; Stable GUS expression
- 刊名：Applied Biochemistry and Biotechnology
- 出版年：2014
- 出版时间：February 2014
- 年：2014
- 卷：172
- 期：4
- 页码：1807-1817
- 全文大小：381 KB
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Zhenrui Gao (1) (3)
Chunlan Piao (1)
Kaiwen Lu (2)
Zhiping Wang (2)
Min-Long Cui (1)
1. Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, 110016, People’s Republic of China
3. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China
2. Yunnan Academy of Applied Technology, Kunming, 650051, People’s Republic of China - ISSN：1559-0291
文摘
In this study, we developed a rapid and efficient method for in vitro propagation and Agrobacterium tumefaciens-mediated transformation of Digitalis purpurea L. (syn. foxglove), an important medicinal plant. Mature leaf explants of D. purpurea were used for 100?% adventitious shoot regeneration on Murashige and Skoog (MS) medium supplemented with 1?mg?L? thidiazuron (TDZ) (a cytokine) and 0.1?mg?L? 1-naphthaleneacetic acid (NAA) (an auxin). Transformation was achieved by inoculating leaf explants with the A. tumefaciens strains GV2260/pBI121 or GV3101/pBI121. The binary vector pBI121 contained the reporter β-glucuronidase gene (GUS) and kanamycin selection marker nptII. Kanamycin-resistant shoots were regenerated directly on the selection medium 4-?weeks after co-cultivation. Approximately, 52.2 and 60?% of kanamycin-resistant shoots transformed with Agrobacterium strains GV2260 and GV3101, respectively, showed strong GUS staining by histochemical assay. Furthermore, PCR and Southern blot analysis confirmed the presence of nptII and GUS on the chromosome of the transformed D. purpurea plants, and stable GUS expression was detected in the transformants by RT-PCR analysis. This efficient method of shoot regeneration and genetic transformation of D. purpurea will provide a powerful tool to increase and produce valuable components such as digitoxin, digoxin, and digoxigenin in D. purpurea through improved secondary metabolic pathways via a biotechnological approach.