In vitro micropropogation of Stevia rebaudiana in Bihar

K RASHMI; S SAHAY; H KUMAR

doi:10.56093/ijas.v89i4.88863

Authors

K RASHMI Scientist, Plant Breeding and Genetics, Rajendra Agricultural University, Pusa, Bihar 848 125, India
S SAHAY Senior Scientist, Fruit Science, Rajendra Agricultural University, Pusa, Bihar 848 125, India
H KUMAR 3Department of Genetics and Molecular Biology, Rajendra Agricultural University, Pusa, Bihar 848 125.

DOI:

https://doi.org/10.56093/ijas.v89i4.88863

Keywords:

Micropropagation, Rebaudioside, Stevia rebaudiana, Stevioside, Tissue culture

Abstract

The shoot apex, leaf, nodal stem and internodal stem of Stevia rebaudiana (Bert.) Bertoni were cultured on 21 different media, having different concentrations and combinations of auxins (2,4-D, IBA, NAA) and cytokinins (KIN, BA) supplemented to Murashige and Skoog (MS) basal medium. Besides these explants, the formed callus and developed shoots were also cultured. The tissue culture experiments resulted in establishment of cultures, swelling of explants, elongation of existing shoot, callus formation, caulogenesis, rhizogenesis and regeneration of plantlets, acclimatization of plantlets and ultimately development of micropropagation protocol. During acclimatization, the plantlets regained their photosynthetic efficiency and transpiration mechanism. The size of leaf also increased during acclimatization. In the present investigation a survival rate of 48% was found during hardening. Good callus formation and differentiation of shoots from formed callus showed the possibility of plant improvement through tissue culture.

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References

Ahmed M B, Salahin M, Karim R, Razvy M A, Hannan M M, Sultana R, Hossain M and Islam R. 2007. An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana Bertoni.) in Bangladesh. American- Eurasian J. Sci. Res. 2(2): 121–5.

Brandle J E and Rosa N. 1992. Heritability for yield, leaf: stem ratio and stevioside content estimated from a landrace cultivar of Stevia rebaudiana. Canadian Journal of Plant Science 72: 1263–6. DOI: https://doi.org/10.4141/cjps92-159

Chalapathi M V and Thimmegowda S. 1997. Natural non-calorie sweetener stevia (Stevia rebaudiana Bertoni) A future crop of India. Crop Research-Hisar 14(2): 347–50.

Chalapathi M V, Thimmegowda S, Sridhara S, Parama V R R and Prasad T G. 1997. Natural non-calorie sweetener stevia (Stevia rebaudiana Bertoni) – A future crop of India. Crop Research 14: 347–50.

Choi Y H, Kim I, Yoon K D, Lee S J, Kim C Y and Yoo K P. 2002. Supercritical fluid extraction and liquid chromatographic-electrospray mass spectrometric analysis of stevioside from Stevia rebaudiana leaves. Chromatographia 55(9–10): 617−20. DOI: https://doi.org/10.1007/BF02492911

Felippe G M and Lucas N M C. 1971. Estudo da viabilidade dos fructos de Stevia rebaudiana Bertoni. Hoehnea 1: 95–105.

Ferreira C M and Handro W. 1988. Micropropagation of Stevia rebaudiana through leaf explants from adult plants. Planta Medica 54: 57–160. DOI: https://doi.org/10.1055/s-2006-962377

Jain P, Kachhwaha S and Kothari S L. 2009. Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana Bertoni by using high copper levels in the culture medium, Scientia Horticulturae 119: 315–9. DOI: https://doi.org/10.1016/j.scienta.2008.08.015

Mitra A and Pal A. 2007. In vitro regeneration of Stevia rebaudiana Bertoni from nodal explants. Journal of Plant Biochemistry and Plant Biotechnology 16: 59–62. DOI: https://doi.org/10.1007/BF03321930

Miyagawa H, Fujioka N, Kohda H, Yamasaki K, Taniguchi K and Tanaka R. 1986. Studies on the tissue culture of Stevia rebaudiana and its components; (II). Induction of shoot primordia. Planta Medica 52: 321–3. DOI: https://doi.org/10.1055/s-2007-969165

Miyazaki Y and Wantenabe H. 1974. Studies on the cultivation of Stevia rebaudiana Bertoni and on the propagation of the plant. Japanese Journal of Tropical Agriculture 17: 154–7.

Mizutani K and Tanaka O. 2002. Use of Stevia rebaudiana sweeteners in Japan. (In) Stevia, the Genus Stevia. A D Kinghorn (Ed), Vol. 19, pp 178–95.

Medicinal and Aromatic Plants Industrial Profile, Taylor and Francis, London.

Murashige T and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissues cultures. Physiologia Plantarum 15: 431–97. DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Patil V, Reddy P C, Purushotham M G, Prasad T G and Udayakumar M. 1996. In vitro multiplication of Stevia rebaudiana Bertoni. Current Science 70: 960.

Razak U N A A, Ong C B , Yu T S and Lau L K. 2014. In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia. Brazilian Archives of Biology and Technology 57(1): 23–8. DOI: https://doi.org/10.1590/S1516-89132014000100004

Sairkar P, Chandravanshi M K, Shukla N P and Mehrotra N N. 2009. Mass production of an economically important medicinal plant Stevia rebaudiana Bertoni using in vitro propagation techniques. Journal of Medicinal Plants Research 3: 266–70.

Shock C C. 1982. Rebaudi’s stevia: Natural non-caloric sweeteners. California Agriculture. 36(9): 4–5.

Sivaram L and Mukundan U. 2003. In vitro culture studies on Stevia rebaudiana. In Vitro Cellular and Development Biology. Plant 39(5): 520–3. DOI: https://doi.org/10.1079/IVP2003438

Starratt A N, Kirby C W, Pocsa R and Brandle J E. 2002. Rebaudioside Fa diterpene glycoside from Stevia rebaudiana. Phytochemistry. 59: 367–70. DOI: https://doi.org/10.1016/S0031-9422(01)00416-2

Tamura Y, Nakamura S, Fukui H and Tabata M. 1984. Clonal propagation of Stevia rebaudiana Bertoni by stem tip culture. Plant Cell Reports 3: 183–5. DOI: https://doi.org/10.1007/BF00270195