A Novel Protocol for Stevia rebaudiana (Bert.) Regeneration

Authors

  • Akram Mirniam Biology Department, Faculty of Sciences, Shahrekord University, Shahrekord, 88186-34141, PO BOX 115, Iran.
  • Parto Roshandel Biology Department, Faculty of Sciences, Shahrekord University, Shahrekord, 88186-34141, PO BOX 115, Iran.
  • Mahmoud Otroshi Agricultural Biotechnology Research Institute of Iran, Isfahan, Iran.
  • Morteza Ebrahimi Agricultural Biotechnology Research Institute of Iran, Isfahan, Iran.

Keywords:

Stevia, Organogenesis, Axillary bud, IAA, IBA, BAP, NAA

Abstract

Stevia rebaudiana Bertoni has sweet substances (stevioside) in its leaves that are free of calories and their consumption is beneficial for diabetic patients and is also helpful in high blood pressure also. Because of low capability in seed germination, tissue culture is an appropriate method for propagation of this plant. In the current study, optimization of stevia in vitro cultivation via direct organogenesis with different explants, light intensities and plant hormones has been examined. These treatments included BAP (at 0.5, 1, 1.5 and 2mg/l) in combination with  2,4-D, IBA and NAA (each with concentrations of 0.1, 0.2 and 0.5mg/l) and different light intensities (Dark, 2000, 4000 and 6000 lux). MS was utilized as a basal medium. Results indicated the highest rate of organogenesis (85%) occurred on the axillary buds explants with a medium containing 1.5mg/l BAP + 0.1mg/l NAA under 6000 lux light intensity. Also, the highest range of primary organ per explant (42) with 0.3cm length was achieved at this condition. The most efficient medium for rhizogenesis i.e. 100% root production along with the highest root number (11 with approximately 7.13cm length) was obtained in presence of activated charcoal and 1mg/l of IBA. At the end of rhizogenesis experiments, the plantlet length and node multiplicity were 12.8cm and 7 respectively. Greenhouse cultivation of these plantlets was successful.

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References

Ahmed, M.B., Salahin, M., Karim, R., Razvy, M.A., Hannan, M.M., Sultana, R., Hossain, M. & Islam, R. (2007). An efficient method for in vitro clonal propagation of a newly introduced sweetener plant (Stevia rebaudiana Bertoni.) in Bangladesh. Am-Euras. J. Sci. Res., 2(2): 121-125.

Arteca, R.N. (1996). Plant Growth Substances: Principles and Applications. New York: Chapman & Hall, 332 pp.

Bondarev, N.I., Nosov, A.M. & Kornienko, A.V. (1998). Effects of exogenous growth regulators on callusogenesis and growth of cultured cells of Stevia rebaudiana Bertoni. Russ. J. Plant Physiol., 45: 770–774.

Bondarev, N., Reshetnyak, O. & Nosov, A. (2002). Features of development of Stevia rebaudiana shoots cultivated in the roller bioreactor and their production of steviol glycosides. Planta Med., 68(8): 759–762. https://doi.org/10.1055/s-2002-33809.

Felippe, G.M. & Lucas, N.M.C. (1971). Estudo da viabilidade dos frutos de Stevia rebaudiana Bert. Hoehnea, 1: 95–105.

Felippe, G.M., Lucas, N.M.C., Behar, L. & Oliveira, M.A.C. (1971). Observações a respeito da germinação de Stevia rebaudiana Bert. Hoehnea, 1: 81-93.

Ferreira, C.M. & Handro, W. (1988). Production, maintenance and plant regeneration from cell suspension cultures of Stevia rebaudiana (Bert.) Bertoni. Plant Cell Rep., 7(2): 123–126. https://doi.org/10.1007/BF00270120.

Handro, W., Hell, K.G. & Kerbauy, G.B. (1977). Tissue culture of Stevia Rebaudiana, a sweetening plant. Planta Med., 32(06): 115–117. https://doi.org/10.1055/s-0028-1097568.

Ibrahim, I.A., Nasr, M.I., Mohammed, B.R. & El-Zefzafi, M.M. (2008). Plant growth regulators affecting in vitro cultivation of Stevia rebaudiana. Sugar Tech., 10(3): 254–259. https://doi.org/10.1007/s12355-008-0045-6.

Jeppesen, P.B., Gregersen, S., Rolfsen, S.E., Jepsen, M., Colombo, M., Agger, A., Xiao, J., Kruhøffer, M., Orntoft, T. & Hermansen, K. (2003). Antihyperglycemic and blood pressure-reducing effects of stevioside in the diabetic Goto-Kakizaki rat. Metabolism, 52(3): 372–378. https://doi.org/10.1053/meta.2003.50058.

Miyagawa, H., Fujioka, N., Kohda, H., Yamasaki, K., Taniguchi, K. & Tanaka, R. (1986). Studies on the tissue culture of Stevia rebaudiana and its components; (II). Induction of shoot primordia. Planta Med., 52(4): 321-323. https://doi.org/10.1055/s-2007-969165.

Akita, M., Shigeoka, T., Koizumi, Y. & Kawamura, M. (1994). Mass propagation of shoots of Stevia rebaudiana using a large scale bioreactor. Plant Cell Rep., 13(3-4): 180–183. https://doi.org/10.1007/BF00239888.

Debnath, M. (2007). Clonal propagation and antimicrobial activity of an endemic medicinal plant Stevia rebaudiana. J. Med. Plant Res., 2(2): 45–51. https://doi.org/10.5897/JMPR.9000368.

Nakamura, S. & Tamura, Y. (1985). Variation in the main Glycosides of Stevia (Stevia rebaudiana Bertoni). Jpn. J. Trop. Agr., 29(2): 109–115. https://doi.org/10.11248/jsta1957.29.109.

Aksenova, N.P., Konstantinova, T.N., Sergeeva, L.I., Macháčková, I. & Golyanovskaya, S.A. (1994). Morphogenesis of potato plants in vitro. I. Effect of light quality and hormones. J. Plant Growth Regul., 13(3): 143–146. https://doi.org/10.1007/BF00196378.

Jain, P., Kachhwaha, S. & Kothari, S.L. (2009). Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana (Bert.) Bertoni by using high copper levels in the culture medium. Sci. Hortic., 119(3): 315–319. https://doi.org/10.1016/j.scienta.2008.08.015.

Rafiq, M., Dahot, M.U., Mangrio, S.M., Naqvi, H.A. & Qarshi, I.A. (2007). In Vitro clonal propagation and biochemical analysis of field established Stevia rebaudiana Bertoni. Pak. J. Bot., 39(7): 2467-2474.

Sakaguchi, M. & Kan, T. (1982). Japanese researches on Stevia rebaudiana (Bert.) Bertoni and stevioside. Ci. Cult., 34: 235–248.

Sivaram, L. & Mukundan, U. (2003). In vitro culture studies on Stevia rebaudiana. In Vitro Cell. Dev. Biol. Plant, 39(5): 520–523. https://doi.org/10.1079/IVP2003438.

Soejarto, D.D., Kinghorn, A.D. & Farnsworth, N.R. (1982). Potential sweetening agents of plant origin. III. Organoleptic evaluation of Stevia leaf herbarium samples for sweetness. J. Nat. Prod., 45(5): 590–599. https://doi.org/10.1021/np50023a013.

Sreedhar, R.V., Venkatachalam, L., Thimmaraju, R., Bhagyalakshmi, N., Narayan, M.S. & Ravishankar, G.A. (2008). Direct organogenesis from leaf explants of Stevia rebaudiana and cultivation in bioreactor. Biol. Plant., 52(2): 355–360. https://doi.org/10.1007/s10535-008-0073-9.

Tamura, Y., Nakamura, S., Fukui, H. & Tabata, M. (1984). Clonal propagation of Stevia rebaudiana Bertoni by stem-tip culture. Plant Cell Rep., 3(5): 183–185. https://doi.org/10.1007/BF00270195.

Tanaka, O. (1982). Steviol-glycosides: New natural sweeteners. TrAC, Trends Anal. Chem., 1(11): 246–248. https://doi.org/10.1016/0165-9936(82)80079-4.

Toffler, F. & Orio, O.A. (1981). Acceni sulla pin ata tropicale ‘Kaa-he-e’ ou ‘erba dolce’. Rev. Soc. Sci. Aliment., 4: 225–230.

Uddin, M.S., Chowdhury, M.S.H., Khan, 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: 1238–1240.

Waller, F. & Nick, P. (1997). Response of actin microfilaments during phytochrome-controlled growth of maize seedlings. Protoplasma, 200(3): 154–162. https://doi.org/10.1007/BF01283291.

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Published

2013-01-01

How to Cite

Mirniam, A., Roshandel, P., Otroshi, M., & Ebrahimi, M. (2013). A Novel Protocol for Stevia rebaudiana (Bert.) Regeneration. Advances in BioScience, 4(1), 15–22. Retrieved from https://journals.sospublication.co.in/ab/article/view/119

Issue

Vol. 4 No. 1 (2013): January

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