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Indirect regeneration of the Cancer bush (Sutherlandia frutescens L.) and detection of l-canavanine in in vitro plantlets using NMR

  • Plant Tissue Culture
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Abstract

The present study reports a simple protocol for indirect shoot organogenesis and plant regeneration of Sutherlandia using rachis and stem segments. Different concentrations (0.0–68.08 μmol l−1) of thidiazuron (TDZ) were used for callus induction and shoot organogenesis. The highest percentage of callus formation (97.5%) and the highest percentage of explants forming shoots (88.8%) were obtained from rachis explants cultured onto Murashige and Skoog (MS) medium (Murashige and Skoog, Physiol. Plant. 15:473–495, 1962) supplemented with 45.41 μmol l−1 TDZ. Scanning electron microscopy demonstrated the early development of adventitious shoots derived from callus cultures. Shoot clusters were further developed and grown in MS hormone-free medium. The presence of l-canavanine was determined by thin-layer chromatography and confirmed after column fractionation using silica gel and nuclear magnetic resonance spectroscopy. Individual shoots were rooted on different concentrations and combinations of MS salt strength and IBA. Half-strength MS salt medium supplemented with 24.6 μmol l−1 IBA was optimal for root induction in which 78% of shoots were rooted. The in vitro plants were successfully acclimatized in a growth chamber with a 90% survival rate.

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References

  • Abrie A. L.; Van Staden J. Micropropagation of the endangered Aloe polyphylla. Plant Growth Regul 33: 19–23; 2001.

    Article  CAS  Google Scholar 

  • Barna K. S.; Wakhlu A. K. Effect of thidiazuron on micropropagation of rose. In Vitro Cell Dev Biol Plant 31: 44–45; 1995.

    Article  CAS  Google Scholar 

  • Burcher S. Can traditional medicine help AIDS? ISIS Press. http://www.i-sis.org.uk/CTMHAids.php. Cited April 1; 2004.

  • Chadwick W. A.; Roux S.; Van de Venter M.; Louw J.; Oelofsen W. Antidiabetic effects of Sutherlandia frutescens in Wistar rats fed a diabetogenic diet. J Ethnopharmacol 109: 121–127; 2007.

    Article  PubMed  Google Scholar 

  • Fracro F.; Echeverrigaray S. Micropagation of Cunila galoides, a popular medicinal plant of South Brazil. Plant Cell Tissue Organ Cult 64: 1–4; 2001.

    Article  Google Scholar 

  • Garland P.; Stoltz L. P. Micropropagation of Pissardi plum. Ann Bot 48: 387–389; 1981.

    Google Scholar 

  • Hartnett S. M.; Oosthuizen V.; Van de Venter M. Anti-HIV activities of organic and aqueous extracts of Sutherlandia frutescens and Lobostemon trigonus. J Ethnopharmacol 96: 113–119; 2005.

    Article  Google Scholar 

  • Huetteman C. A.; Preece J. E. Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33: 105–119; 1993.

    Article  CAS  Google Scholar 

  • Hutchinson M. J.; Saxena P. K. Acetylsalicylic acidenhances and synchronizes thidiazuron-induced somatic embryogenesis in geranium (Pelargonium × hortorum Bailey) tissue culture. Plant Cell Rep 15: 512–515; 1996.

    Article  CAS  Google Scholar 

  • Katerere D. R.; Eloff J. N. Antibacterial and antioxidant activity of Sutherlandia frutescens (Fabaceae), a reputed anti-HIV/AIDS phytomedicine. Phytother Res 19: 779–781; 2005.

    Article  PubMed  Google Scholar 

  • Kundu J. K.; Mosanda K. S.; Na H.; Surh Y. J. Inhibitory effects of the extracts of Sutherlandia frutescens (L.) R. Br. and Harpagophytum procumbens DC. on phorbol ester-induced COX-2 expression in mouse skin: AP-1 and CREB as potential upstream targets. Cancer Lett 218: 21–31; 2005.

    Article  CAS  PubMed  Google Scholar 

  • Liu C. Z.; Murch S. J.; Demerdash E. L.; Saxena P. K. Regeneration of the Egyptian medicinal plant Artemisia judaica L. Plant Cell Rep 21: 525–530; 2003.

    CAS  PubMed  Google Scholar 

  • Manzanera J. A.; Pardos J. A. Micropropagation of juvenile and adult Quercus suber. Plant Cell Tissue Organ Cult 21: 1–8; 1990.

    Article  CAS  Google Scholar 

  • Mithila J.; Hall J. C.; Victor J. M. R.; Saxena P. K. Thidiazuron induces shoot organogenesis at low concentrations and somatic embryogenesis at high concentrations on leaf and petiole explants of African violet (Saintpauliaionantha Wendl.). Plant Cell Rep 21: 408–414; 2003.

    CAS  PubMed  Google Scholar 

  • Murashige T.; Skoog F. A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15: 473–495; 1962.

    Article  CAS  Google Scholar 

  • Murthy B. N. S.; Murch S. J.; Saxena P. K. Thidiazuron induced somatic embryogenesis in intact seedlings of peanut (Arachis hypogaea L.): endogenous growth regulator levels and significance of cotyledons. Physiol Plant 94: 268–276; 1995.

    Article  CAS  Google Scholar 

  • Murthy B. N. S.; Murch S. J.; Saxena P. K. Thidiazuron: a potential regulator of in vitro plant morphogenesis. In Vitro Cell Dev Biol Plant 34: 267–275; 1998.

    Article  CAS  Google Scholar 

  • Paek K. Y.; Chakrabarty D.; Hahn E. J. Application of bioreactor system for large scale production of horticultural and medicinal plants. Plant Cell Tissue Organ Cult 81: 287–300; 2005.

    Article  Google Scholar 

  • Purohit S. D.; Kukda G.; Sharma P.; Tak K. In vitro propagation of an adult tree Wrightia lomentosa through enhanced axilliary branching. Plant Sci 103: 67–72; 1994.

    Article  CAS  Google Scholar 

  • Reid K. A.; Maes J.; Maes A.; Van Staden J.; De Kimpe N.; Mulholland D. A.; Verschaeve L. Evaluation of the mutagenic and antimutagenic effects of South African plants. J Ethnopharmacol 106: 44–50; 2006.

    Article  CAS  PubMed  Google Scholar 

  • Schween G.; Schwenkel H. G. In vitro regeneration in Primula ssp. via organogenesis. Plant Cell Rep 20: 1006–1010; 2002.

    Article  CAS  Google Scholar 

  • Smetanska I.; Riedel H. Production of phenolics from medicinal plants by cell cultures. European Society for New Methods in Agricultural Research—XXXVII Annual Meeting, Dubna, Russia, Poster Congress Sept. 10–14; 2007

  • Shaik S.; Dewir Y. H.; Singh N.; Nicholas A. Influences of pre-sowing seed treatments on germination of the cancer bush (Sutherlandia frutescens), a reputed medicinal plant in arid environments. Seed Sci Technol 36: 794–800; 2008.

    Google Scholar 

  • Singh N.; Potukuchi S. M. Phytocompound production and evaluation in in vitro culture systems. 11th IAPTC&B Congress, Beijing, China; 2006

  • Stander B. A.; Marais S.; Steynberg T. J.; Theron D.; Joubert F.; Albrecht C.; Joubert A. M. Influence of Sutherlandia frutescens extracts on cell numbers, morphology and gene expression in MCF-7 cells. J Ethnopharmacol 112: 312–318; 2007.

    Article  CAS  PubMed  Google Scholar 

  • Tao J.; Qin C.; Yao L.; Jiang W.; Zhan Y.; Li Y. The study on the effect of different factors on in vitro rooting of regenerated plantlets for Betula platyphylla. JiLin Forest Sci Technol 2: 1–8; 1999.

    Google Scholar 

  • Thomas T. D. Thidiazuron induced multiple shoot induction and plant regeneration from cotyledonay explants of mulberry. Biol Plant 46: 529–533; 2003.

    Article  CAS  Google Scholar 

  • Trigiano R. N.; Gray D. J. Plant tissue culture: concepts and laboratory exercises. 2nd ed. CRC, Boca Raton; 2000.

    Google Scholar 

  • Van Wyk B. B.; Van Oudtshoorn B.; Gericke N. Medicinal plants of South Africa. Briza Publications, Pretoria; 1997.

    Google Scholar 

  • Watt J. M.; Breyer-Brandwijk M. G. The medicinal and poisonous plants of southern and eastern Africa. 2nd ed. Livingstone, London; 1962.

    Google Scholar 

  • Zimmerman R. H.; Broome O. C. Phloroglucinol and in vitro rooting of apple cultivar cuttings. J Am Soc Hort Sci 106: 648–652; 1981.

    CAS  Google Scholar 

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Acknowledgment

The University of KwaZulu-Natal and the National Research Foundation (NRF) are thanked for financial support. The authors wish to thank Prof. David Mycock for his help in scanning electron microscopy.

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Correspondence to Yaser Hassan Dewir.

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Editor: R. A. Shibli

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Dewir, Y.H., Singh, N., Shaik, S. et al. Indirect regeneration of the Cancer bush (Sutherlandia frutescens L.) and detection of l-canavanine in in vitro plantlets using NMR. In Vitro Cell.Dev.Biol.-Plant 46, 41–46 (2010). https://doi.org/10.1007/s11627-009-9260-4

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  • DOI: https://doi.org/10.1007/s11627-009-9260-4

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