Skip to main content
SpringerLink
Log in

Optimization of induction and culture conditions and tropane alkaloid production in hairy roots of Anisodus acutangulus

  • Published:
Biotechnology and Bioprocess Engineering Aims and scope Submit manuscript

Abstract

In this study, an efficient transformation system for the medicinal plant Anisodus acutangulus was successfully developed and optimized using Agrobacterium rhizogenes. Three bacterial strains, A4, R1601, and modified C58C1 and three explant types, leaf blade, petiole, and stem, were examined. The highest transformation efficiency of 94.44% was achieved using strain C58C1 with stem explants. Over 20 independent hairy root lines were successfully established with strain C58C1 using stem explants, all of which contained the ro/B and ro/C genes as confirmed by polymerase chain reaction (PCR). Out of four media compositions, the liquid 1/2 MS medium was found the most suitable for hairy root growth. The maximum biomass of one hairy root line increased up to 80 times in liquid 1/2 MS medium after a 30 day culture period. Different hairy root lines displayed a varied capacity for tropane alkaloid production and the best hairy root line (T4) from the C58C1-stem combination produced up to 10.21 mg/g (dw) of hyoscyamine, which was about 1.5-fold higher than in the wild type plants. To our knowledge, this is the first report to demonstrate the production of tropane alkaloids in hairy roots of A. acutangulus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Xiao, P. G., G. C. Xia, and L. Y. He (1973) The occurrence of some important tropane alkakoids in Chinese solanaceous pants. Acta Bot. Sin. 15: 187–194.

    Google Scholar 

  2. Kai, G., J. Chen, L. Li, G. Zhou, L. Zhou, L. Zhang, Y. Chen, and L. Zhao (2007) Molecular cloning and characterization of a new cDNA encoding hyoscyamine 6β-hydroxylase from roots of Anisodus acutangulus. J. Biochem. Mol. Biol. 40: 715–722.

    CAS  Google Scholar 

  3. Kang, S. M., H. Y. Jung, Y. M. Kang, D. J. Yun, J. D. Bahk, J. K. Yang, and M. S. Choi (2004) Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Sci. 166: 745–751.

    Article  CAS  Google Scholar 

  4. Huang, F., X. D. Dai, Y. L. Hu, C. Y. Chen, and G. Z. Zhu (2005) Progress in synthesis of tropane alkaloids. Chem. Reagent 27: 141–144.

    Google Scholar 

  5. Yun, D. J., T. Hashimoto, and Y. Yamada (1992) Metabolic engineering of medicinal plants: transgenic Atropa belladonna with an improved alkaloid composition. Proc. Natl. Acad. Sci. USA 89: 11799–11803.

    Article  CAS  Google Scholar 

  6. Zheng, G. Z., J. B. He, and S. L. Wang (1982) Cell suspension culture of callus from Anisodus acutangulus roots. Acta Botanica Yunnanca 4: 203–206.

    Google Scholar 

  7. Canto-Canche, B. and V. M. Loyola-Vargas (1999) Chemicals from roots, hairy roots, and their application. pp. 235–275. In: F. Shahidi, P. Kolodziejczyk, J. R. Whitaker, A. L. Munguia, and G. Fuller (eds.). Chemicals via Higher Plant Bioengineering (Advances in Experimental Medicine and Biology, 464). Kluwer Academic Publishers, New York, NY, USA.

    Google Scholar 

  8. Sevon, N. and K. M. Oksman-Caldentey (2002) Agrobacterium rhizogenes-mediated transformation: root cultures as a source of alkaloids. Planta Med. 68: 859–868.

    Article  CAS  Google Scholar 

  9. Tiwari, R. K., M. Trivedi, Z. C. Guang, G. Q. Guo, and G. C. Zheng (2007) Genetic transformation of Gentiana macrophylla with Agrobacterium rhizogenes: growth and production of secoiridoid glucoside gentiopicroside in transformed hairy root cultures. Plant Cell Rep. 26: 199–210.

    Article  CAS  Google Scholar 

  10. Zeng, Y. L. (1962) Pharmaceutical study and investigation of Anisodus acutangulus in Yunnan. Med. Pharm. Yunnan 3: 64–66.

    Google Scholar 

  11. Wu, D. K., F. L. Wang, Z. R. Chen, J. S. Yang, and Q. L. Huang (1962) Chemical analysis of Anisodus acutangulus in Yunnan and extraction of atropine sulphate. Med. Pharm. Yunnan 3: 67–68.

    Google Scholar 

  12. Meng, C., X. Zuo, L. Wang, P. Zhu, and K. D. Cheng (2002) Production of scopolamine by hairy root cultures in Anisodus tanguticus. Nat. Prod. Res. Dev. 14: 21–24.

    CAS  Google Scholar 

  13. Kamada, H., N. Okamura, M. Satake, H. Harada, and K. Shimomura (1986) Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Rep. 5: 239–242.

    Article  CAS  Google Scholar 

  14. Ondrej, M. and J. Protiva (1987) In vitro culture of crown gall and hairy root tumors of Atropa belladonna: Differentiation and alkaloid production. Biol. Plant. 29: 241–246.

    Article  CAS  Google Scholar 

  15. Jung, G. and D. Tepfer (1987) Use of genetic transformation by the Ri T-DNA of Agrobacterium rhizogenes to stimulate biomass and tropane alkaloid production in Atropa belladonna and Calystegia sepium roots. Plant Sci. 50: 145–151.

    Article  CAS  Google Scholar 

  16. Christen, P., M. F. Roberts, J. D. Phillipson, and W. C. Evans (1989) High-yield production of tropane alkaloids by hairy-root cultures of a Datura candida hybrid. Plant Cell Rep. 8: 75–77.

    Article  CAS  Google Scholar 

  17. Payne, J., J. D. Hamill, R. J. Robins, and M. J. C. Rhodes (1987) Production of hyoscyamine by ‘hairy root’ cultures of Datura stramonium. Planta Med. 53: 474–478.

    Article  CAS  Google Scholar 

  18. Jaziri, M., M. Legros, J. Homes, and M. Vanhaelen (1988) Tropine alkaloids production by hairy root cultures of Datura stramonium and Hyoscyamus niger. Phytochemistry 27: 419–420.

    Article  CAS  Google Scholar 

  19. Mano, Y., S. Nabeshima, C. Matsui, and H. Ohkawa (1986) Production of tropane alkaloids by hairy root cultures of Scopolla japonica. Agric. Biol. Chem. 50: 2715–2722.

    CAS  Google Scholar 

  20. Pan, X. C., M. Chen, L. Zhang, Z. H. Liao, and M. S. Zhang (2007) Efficient expression of plant expressing vector in Anisodus acutangulus hairy roots. Chin. Tradit. Herb. Drugs 38: 588–591.

    CAS  Google Scholar 

  21. Doyle, J. J. and J. L. Doyle (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13–15.

    Google Scholar 

  22. Sevón, N., K. M. Oksman-Caldentey, and R. Hiltunen (1995) Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus. Plant Cell Rep. 14: 738–742.

    Article  Google Scholar 

  23. Sevón, N., B. Dräger, R. Hiltunen, and K. M. Oksman-Caldentey (1997) Characterization of transgenic plants derived from hairy roots of Hyoscyamus muticus. Plant Cell Rep. 16: 605–611.

    Article  Google Scholar 

  24. Maldonado-Mendoza, I. E., T. Ayora-Talavera, and V. M. Loyola-Vargas (1993) Establishment of hairy root cultures of Datura stramonium. Plant Cell Tissue Organ Cult. 33: 321–329.

    Article  CAS  Google Scholar 

  25. Dhakulkar, S., T. R. Ganapathi, S. Bhargava, and V. A. Bapat (2005) Induction of hairy roots in Gmelina arborea Roxb. and production of verbascoside in hairy roots. Plant Sci. 169: 812–818.

    Article  CAS  Google Scholar 

  26. Moyano, E., K. Jouhikainen, P. Tammela, J. Palazon, R. M. Cusido, M. T. Pinol, T. H. Teeri, and K. M. Oksman-Caldentey (2003) Effect of pmt gene overexpression on tropane alkaloid production in transformed root cultures of Datura metel and Hyoscyamus muticus. J. Exp. Bot. 54: 203–211.

    Article  CAS  Google Scholar 

  27. Nilsson, O. and O. Olsson (1997) Getting to the root: the role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots. Physiol. Plant. 100: 463–473.

    Article  CAS  Google Scholar 

  28. Sauerwein, M., M. Wink, and K. Shimomura (1992) Influence of light and phytohormones on alkaloid production in transformed root cultures of Hyoscyamus albus. J. Plant Physiol. 140: 147–152.

    CAS  Google Scholar 

  29. Hashimoto, T., D. J. Yun, and Y. Yamada (1993) Production of tropane alkaloids in genetically engineered root cultures. Phytochemistry 32: 713–718.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University, Shanghai, 200234, China

    Li Li, Jing Wang, Wei Wang, Yang Lu, Genyu Zhou & Guoyin Kai

  2. Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai, 200030, China

    Yuliang Wang

Authors
  1. Li Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yang Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yuliang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Genyu Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guoyin Kai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guoyin Kai.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, L., Wang, J., Wang, W. et al. Optimization of induction and culture conditions and tropane alkaloid production in hairy roots of Anisodus acutangulus . Biotechnol Bioproc E 13, 606–612 (2008). https://doi.org/10.1007/s12257-008-0035-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12257-008-0035-2

Keywords

Search

Navigation