Regenerative callus induction and biochemical analysis of Stevia rebaudiana Bertoni

Authors

  • Dhurva P. Gauchan Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Ashna Dhakal Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Nisha Sharma Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Sabin Bhandari Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Elina Maskey Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Nayan Shrestha Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Rachita Gautam Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Sarala Giri Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.
  • Sushma Gurung Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel 45200, Nepal.

Keywords:

Stevia rebaudiana, callus, 2,4-D, phytochemical screening, plant growth regulators (PGR)

Abstract

Stevia Leaves are the principal source of stevioside, which is estimated to be 100-300 times sweeter than table sugar. Stevioside has clinical significance as they are reported to maintain glucose levels in human blood. Owing to the difficulties in propagation of stevia through seeds and vegetative methods, callus culture has been an efficient alternative for generation of stevioside. The aim of this study is to develop an efficient and standardized protocol for maximum induction and multiplication of callus from a leaf. Callus culture was established from leaves in MS basal media fortified with various combinations (BAP, NAA, 2,4-D, KN, IBA) and concentrations of phytohormones. The best callusing (100%) was recorded in MS media supplemented with (2,4-D 1.0mg/l + NAA 1.0mg/l). The callus was harvested after 4 weeks and screened for the presence of various bioactive compounds. The qualitative results showed that the extracts of callus contained bioactive compounds like flavonoids, glycosides, phenol, tannins, sterols and saponins thereby making callus one of the sources for extraction of various secondary metabolites.

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References

Abou-Arab, A.E., Abou-Arab, A.A. & Abu-Salem, M.F. (2010). Physico-chemical assessment of natural sweeteners steviosides produced from Stevia rebaudiana bertoni plant. Afr. J. Food Sci., 4(5): 269–281. https://doi.org/10.5897/AJFS.9000226.

Anbazhagan, M., Kalpana, M., Rajendran, R., Natarajan, V. & Dhanavel, D. (2010). In vitro production of Stevia rebaudiana Bertoni. Emir. J. Food Agric., 22(3): 216–222. https://doi.org/10.9755/ejfa.v22i3.4891.

Chan, L.K., Dewi, P.R. & Boey, R.L. (2005). Effect of plant growth regulators on regeneration of plantlets from bud cultures of Cymbopogon nardus L. and the detection of essential oils from the in vitro plantlets. J. Plant Biol., 48(1): 142–146. https://doi.org/10.1007/BF03030574.

Curi, R., Alvarez, M., Bazotte, R.B., Botion, L.M., Godoy, J.L. & Bracht, A. (1986). Effect of Stevia rebaudiana on glucose tolerance in normal adult humans. Braz. J. Med. Biol. Res., 19(6): 771–774.

Das, A., Biswas, M. & Mandal, N. (2010). An economic analysis of Stevia (Stevia rebaudiana Bert.) cultivation through stem cutting and tissue culture propagule in India. Trends in Agricultural Economics, 3(4): 216-222. https://dx.doi.org/10.3923/tae.2010.216.222.

Das, K., Dang, R. & Rajasekharan, P.E. (2006). Establishment and maintenance of callus of Stevia rebaudiana Bertoni under aseptic environment. Natural Product Radiance, 5(5): 373-376.

Ferreira, C.M. & Handro, W. (1988). Micropropagation of Stevia rebaudiana through leaf explants from adult plants. Planta Med., 54(2): 157–160. https://doi.org/10.1055/s-2006-962377.

Gupta, P., Sharma, S. & Saxena, S. (2010). Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production. International Journal of Biological, Biomolecular, Agricultural, Food and Biotechnological Engineering, 4(12): 893-897.

Guruchandran, V. & Sasikumar, C. (2013). Organogenic plant regeneration via callus induction in Stevia rebaudiana Bert. Int. J. Curr. Microbiol. App. Sci., 2(2): 56-61.

Harborne, J.B. (1973). Phytochemical methods: A guide to modern techniques of plant analysis. Chapman and Hall Ltd., London. pp. 49-188.

Janarthanam, B., Gopalakrishnan, M., Sai, G.L. & Sekar, T. (2009). Plant Regeneration from Leaf Derived Callus of Stevia rebaudiana Bertoni. Plant Tissue Cult. Biotech., 19(2): 133-141. https://doi.org/10.3329/ptcb.v19i2.5430.

Kinghora, A.D., Soejarto, D.D. & Inglett, G.E. (1986). Sweetening agents of plant origin. Crit. Rev. Plant Sci., 4(2): 79–120. https://doi.org/10.1080/07352688609382220.

Naz, S., Hashmi, A. & Ali, A. (2009). In vitro callogenesis and organogenesis in different explants of Stevia (Stevia rebaudiana). Pakistan Sugar Journal, 24(2): 20-31.

Pande, S. & Khetmalas, M. (2012). Biological Effect of sodium azide and colchicine on seed germination and callus induction in Stevia rebaudiana. Asian J. Exp. Biol. Sci., 3(1): 93-98.

Uddin, M.S., Chowdhury, M.S.H., Khan, M.M.M.H., Uddin, M.B., Ahmed, R. & Baten, M.A. (2006). In vitro propagation of Stevia rebaudiana Bert in Bangladesh. Afr. J. Biotechnol., 5(13): 1238–1240.

Upadhyay, S., Sharma, S. & Kumar, R. (2013). In vitro morphological, biochemical and microbial studies on elite clones of Stevia rebaudiana for enhanced production of Stevioside. Intl. J. of Trad. Herb. Med., 1(1): 6-12.

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Published

2014-07-01

How to Cite

Gauchan, D. P., Dhakal, A., Sharma, N., Bhandari, S., Maskey, E., Shrestha, N., Gautam, R., Giri, S., & Gurung, S. (2014). Regenerative callus induction and biochemical analysis of Stevia rebaudiana Bertoni. Advances in BioScience, 5(3), 41–45. Retrieved from https://journals.sospublication.co.in/ab/article/view/159

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