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An improved Agrobacterium tumefaciens mediated transformation of Artemisia annua L. by using stem internodes as explants

Author

Listed:
  • Na Tian

    (Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Natural Products Research Center, College of Horticulture and Landscape, Hunan Agricultural University, Changsha, China)

  • Shuoqian Liu

    (Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Natural Products Research Center, College of Horticulture and Landscape, Hunan Agricultural University, Changsha, China
    Laboratory of Plant Physiology, Wageningen University, Wageningen, The Netherlands)

  • Hiengming Ting

    (Laboratory of Plant Physiology, Wageningen University, Wageningen, The Netherlands)

  • Jianan Huang

    (Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Natural Products Research Center, College of Horticulture and Landscape, Hunan Agricultural University, Changsha, China)

  • Sander Van Der Krol

    (Laboratory of Plant Physiology, Wageningen University, Wageningen, The Netherlands)

  • Harro Bouwmeester

    (Laboratory of Plant Physiology, Wageningen University, Wageningen, The Netherlands)

  • Zhonghua Liu

    (Hunan Provincial Key Laboratory for Germplasm Innovation and Utilization of Crop, Natural Products Research Center, College of Horticulture and Landscape, Hunan Agricultural University, Changsha, China)

Abstract

Transformation of Artemisia annua, which produces the sesquiterpenoid endoperoxide artemisinin widely used for the treatment of malaria, has been hampered by the low efficiency of adventitious shoot and root formation on a selective medium containing additional compounds for Agrobacterium decontamination. Here we identified several factors which were all shown to be of importance for optimization of Artemisia annua transformation. Results indicated that stem internodes showed better resistance capacity to Agrobacterium decontaminator than leaves did. Agrobacterium tumefaciens with an optical density (OD) value of 0.2-0.5 plus 100 μmol of acetosyringone per litre of solution gave the best transformation efficiency. Moreover, kanamycin at 30 mg/l in the culture medium was effective in suppressing the growth of non-transformed tissue. Furthermore, transgenic shoots required an early induction of rooting. In addition, dimethyl sulphoxide considerably improved the rooting of shoots. The present work provides rapid and reproducible transformation and regeneration of A. annua.

Suggested Citation

  • Na Tian & Shuoqian Liu & Hiengming Ting & Jianan Huang & Sander Van Der Krol & Harro Bouwmeester & Zhonghua Liu, 2013. "An improved Agrobacterium tumefaciens mediated transformation of Artemisia annua L. by using stem internodes as explants," Czech Journal of Genetics and Plant Breeding, Czech Academy of Agricultural Sciences, vol. 49(3), pages 123-129.
    • Handle: RePEc:caa:jnlcjg:v:49:y:2013:i:3:id:105-2012-cjgpb
    DOI: 10.17221/105/2012-CJGPB
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    1. Richard Van Noorden, 2010. "Demand for malaria drug soars," Nature, Nature, vol. 466(7307), pages 672-673, August.
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