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A Stable and Efficient Agrobacterium tumefaciens-Mediated Genetic Transformation of the Medicinal Plant Digitalis purpurea L.

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Abstract

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−1 thidiazuron (TDZ) (a cytokine) and 0.1 mg L−1 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 4–6 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.

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Acknowledgments

This work was supported by a grant from Yunnan provincial Science and Technology Department (no. 2012IB001) to MLC.

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Authors and Affiliations

  1. Key Laboratory of Pollution Ecology and Environmental Engineering, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, 110016, People’s Republic of China

    Ying Li, Zhenrui Gao, Chunlan Piao & Min-Long Cui

  2. Yunnan Academy of Applied Technology, Kunming, 650051, People’s Republic of China

    Kaiwen Lu & Zhiping Wang

  3. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Ying Li & Zhenrui Gao

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  1. Ying Li
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  2. Zhenrui Gao
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  3. Chunlan Piao
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  6. Min-Long Cui
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Correspondence to Min-Long Cui.

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Li, Y., Gao, Z., Piao, C. et al. A Stable and Efficient Agrobacterium tumefaciens-Mediated Genetic Transformation of the Medicinal Plant Digitalis purpurea L.. Appl Biochem Biotechnol 172, 1807–1817 (2014). https://doi.org/10.1007/s12010-013-0648-6

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  • DOI: https://doi.org/10.1007/s12010-013-0648-6

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