Czech J. Genet. Plant Breed., 2013, 49(3):123-129 | DOI: 10.17221/105/2012-CJGPB

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

Na Tian1, *, Shuoqian Liu1, 2, *, Hiengming Ting2, Jianan Huang1, Sander Van Der Krol2, Harro Bouwmeester2, Zhonghua Liu1
1 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
2 Laboratory of Plant Physiology, Wageningen University, Wageningen, The Netherlands

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.

Keywords: anti-malaria; artemisinin; stem internode; traditional Chinese medicine; transgene

Published: September 30, 2013  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Tian N, Liu S, Ting H, Huang J, Van Der Krol S, Bouwmeester H, Liu Z. An improved Agrobacterium tumefaciens mediated transformation of Artemisia annua L. by using stem internodes as explants. Czech J. Genet. Plant Breed.. 2013;49(3):123-129. doi: 10.17221/105/2012-CJGPB.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Aquil S., Husaini A.M., Abdin M.Z., Rather G.M. (2009): Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic plants. Planta Medica, 75: 1453-1458. Go to original source... Go to PubMed...
    2. Dutt M., Grosser J.W. (2009): Evaluation of parameters affecting Agrobacterium-mediated transformation of citrus. Plant Cell, Tissue and Organ Culture, 98: 331-340. Go to original source...
    3. Han J.L., Wang H., Ye H.C., Liu Y., Li Z.Q., Zhang Y., Zhang Y.S., Yan F., Li G.F. (2005): High efficiency of genetic transformation and regeneration of Artemisia annua L. via Agrobacterium tumefaciens-mediated procedure. Plant Science, 168: 73-80. Go to original source...
    4. Hocking P.J., Thomas M.B. (1980): Rooting of cuttings of Senecio greyi: effect of IBA in combination with DMSO, benomyl, and captan. New Zealand Journal of Experimental Agriculture, 8: 49-54. Go to original source...
    5. Jefferson R.A., Kavanagh T.A., Bevan M.W. (1987): GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher-plants. EMBO Journal, 6: 3901-3907. Go to original source... Go to PubMed...
    6. Lazo G.R., Stein P.A., Ludwig R.A. (1991): A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Nature Biotechnology, 9: 963-967. Go to original source... Go to PubMed...
    7. Lualon W., De-Eknamkul W., Tanaka H., Shoyama Y., Putalun W. (2008): Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron. Zeitschrift für Naturforschung C, 63: 96-100. Go to original source... Go to PubMed...
    8. Mannan A., Syed T.N., Mirza B. (2009): Factors affecting Agrobacterium tumefaciens mediated transformation of Artemisia absinthium L. Pakistan Journal of Botany, 41: 3239-3246.
    9. Mckinniss G.R. (1969): The effects of DMSO as a carrier for IBA on the rooting of three Juniper species. Plant Propagation, 15: 4-8.
    10. Milhous W.K., Weina P.J. (2010): Plant science. The botanical solution for malaria. Science, 327: 279-280 Go to original source... Go to PubMed...
    11. Praygod G., De Frey A., Eisenhut M. (2008): Artemisinin derivatives versus quinine in treating severe malaria in children: a systematic review. Malaria Journal, 7: 210. Go to original source... Go to PubMed...
    12. Sangwan N.S., Sangwan R.S., Kumar S. (1998): Isolation of genomic DNA from the antimalarial plant Artemisia annua. Plant Molecular Biology Reporter, 16: 365-365. Go to original source...
    13. Van Noorden R. (2010): Demand for malaria drug soars. Nature, 466: 672-673. Go to original source... Go to PubMed...
    14. Vergauwe A., Cammaert R., Vandenberghe D., Genetello C., Inze D., Vanmontagu M., Vandeneeckhout E. (1996a): Agrobacterium tumefaciens-mediated transformation of Artemisia annua L. and regeneration of transgenic plants. Plant Cell Report, 15: 929-933. Go to original source... Go to PubMed...
    15. Vergauwe A., Vangeldre E., Inze D., Vanmontagu M., Vandeneeckhout E. (1996b): The use of amoxicillin and ticarcillin in combination with a beta-lactamase inhibitor as decontaminating agents in the Agrobacterium tumefaciens-mediated transformation of Artemisia annua L. Journal of Biotechnology, 52: 89-95. Go to original source...
    16. Vergauwe A., Van Geldre E., Inze D., Van Montagu M., Van den Eeckhout E. (1998): Factors influencing Agrobacterium tumefaciens-mediated transformation of Artemisia annua L. Plant Cell Report, 18: 105-110. Go to original source...
    17. Whatley B.T., Thompson S.O., Williams G. (1966): The effect of nutrient solution, foliar spray, 3-indolebutyric acid and dimethylsulfoxide on rooting of Hibiscus syriacus. Proceedings of the International Plant Propagator's Society, 16: 287-291.
    18. Zhang L., Jing F., Li F., Li M., Wang Y., Wang G., Sun X., Tang K. (2009): Development of transgenic Artemisia annua (Chinese wormwood) plants with an enhanced content of artemisinin, an effective anti-malarial drug, by hairpin-RNA-mediated gene silencing. Biotechnology and Applied Biochemistry, 52: 199-207. Go to original source... Go to PubMed...

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content