Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production
Stevia rebaudiana Bertoni (natural sweetener) belongs
to Asteraceae family and can be used as substitute of artificial
sweeteners for diabetic patients. Conventionally, it is cultivated by
seeds or stem cutting, but seed viability rate is poor. A protocol for
callus induction and multiplication was developed to produce large
no. of calli in short period. Surface sterilized nodal, leaf and root
explants were cultured on Murashige and Skoog (MS) medium with
different concentrations of plant hormone like, IBA, kinetin, NAA,
2,4-D, and NAA in combination with 2,4-D. 100% callusing was
observed from leaf explants cultured on combination of NAA and
2,4-D after three weeks while with 2,4-D, only 10% callusing was
observed. Calli obtained from leaf and root explants were shiny green
while with nodal explants it was hard and brown. The present
findings deal with induction of callusing in Stevia to achieve the
rapid callus multiplication for study of steviol glycosides in callus
culture.
[1] N.I. Bondarev, A.M. Nosov, A.V. Kornienko, "Effects of exogenous
growth regulators on callusogenesis and growth of cultured cells of
Stevia rebaudiana Bertoni", Russ. J. Plant Physiol. vol. 45, pp. 888-
892, 1998.
[2] P. Gupta, S. Sharma, S. Saxena, "Micropropagation of Stevia
rebaudiana (natural sweetener) using Kinetin for Steviol glycoside
Production", Research journal of Biotechnology, vol. 5, no. 1, pp. 63-
67, 2010.
[3] S. Gurel, E. Gurel, Z. Kaya, "Callus development and indirect shoot
regeneration from seedling explants of sugarbeet (Beta Vulgaris)
cultured in-vitro", Turk. J. Bot, vol. 22, pp. 25-33, 2001.
[4] Y.I. Hsing, W.F. Su, W.C. Chang, "Accumulation of stevioside and
rebaudioside A in callus cultures of Stevia rebaudiana Bertoni", Bot.
Bull. Acad. Sin. vol. 24, pp. 115-119, 1983.
[5] P. Jain, S. Kaxhhwaha, S. L. Kothari, "Improved micropropagation
protocol and enhancement in biomass and chlorophyll content in Stevia
rebaudiana (Bert.) Bertoni by using high copper levels in the culture
medium", Scientia Horticulturae, vol. 119, pp.315-319, 2009.
[6] B. Janarthanam, M. Gopalakrishnan, S G. Lakshmi, T. Sekar, "Plant
regeneration from leaf derived callus of Stevia rebaudiana Bertoni",
Plant Tissue Culture and Biotechnology, vol.19, no. 2, pp. 133-141,
2009.
[7] A.D. Kinghorn, D.D. Soejarto, "Sweetening agents of plant origin".
Crit. Rev. Plant Sci. 4, pp. 79-120, 1986.
[8] F.A. Krens, Jamar. D, "The role of explant source and culture
conditions on callus induction and shoot regeneration in sugarbeet
(Beta vulgaris L.)", J. Plant Physiol, vol. 134, pp. 651-655, 1989.
[9] A.G. Lyakhovkin, T.D. Long, D.A. Titov, M.P. Anh, "In: Cultivation
and utilization of Stevia (Stevia rebaudiana Bertoni)", Agricultural
Publishing House, Hanoi, pp. 1-44, 1993.
[10] R.M. Patel, R. R. Shah, "Regeneration of Stevia Plant through Callus
Culture", Indian Journal of Pharmaceutical Sciences, pp. 46-50, vol.2,
Jan-Feb. 2009.
[11] L. Sivaram, U. Mukundan, "In vitro culture studies on Stevia
rebaudiana", In Vitro Cell. Dev. Biol.-Plant, vol. 39, pp 520-523,
September-October. 2003.
[12] S.M. Swanson, G.B. Mahady, C.W.W. Beecher, "Stevioside
biosynthesis by callus, root, shoot and rooted-shoot cultures in vitro".
Plant Cell Tissue Organ Cult. Vol. 28, pp. 151-157 1992.
[13] M.B. Tadhni, V.H. Patel, R. Subhash, "In vitro antioxidant activities of
Stevia rebaudiana leaves and callus". Journal of food composition and
Analysis , vol.20, pp. 323-329, 2007.
[14] A.S. Taware, D. S. Mukadam, A. M. Chavan, S.D. Taware,
"Comparative study of in-vitro and in-vivo grown plants and callus of
Stevia rebaudiana (Bertoni)", International Journal of integrative
Biology, vol.9, no. 1, pp 10-15, 2010.
[15] MS. Uddin, M. S. Chowdhary, H. Khan, "In vitro propagation of Stevia
rebaudiana Bert in Bangladesh", African Journal of Biotechnology,
vol. 5, no.13, pp. 1238-1240, July. 2006.
[16] T. Yamazaki, H.E. Flores, "Examination of steviol glycoside
production by hair root and shoot cultures of Stevia rebaudiana". J.
Nat. Prod, vol 54, pp. 986-992, 1991.
[1] N.I. Bondarev, A.M. Nosov, A.V. Kornienko, "Effects of exogenous
growth regulators on callusogenesis and growth of cultured cells of
Stevia rebaudiana Bertoni", Russ. J. Plant Physiol. vol. 45, pp. 888-
892, 1998.
[2] P. Gupta, S. Sharma, S. Saxena, "Micropropagation of Stevia
rebaudiana (natural sweetener) using Kinetin for Steviol glycoside
Production", Research journal of Biotechnology, vol. 5, no. 1, pp. 63-
67, 2010.
[3] S. Gurel, E. Gurel, Z. Kaya, "Callus development and indirect shoot
regeneration from seedling explants of sugarbeet (Beta Vulgaris)
cultured in-vitro", Turk. J. Bot, vol. 22, pp. 25-33, 2001.
[4] Y.I. Hsing, W.F. Su, W.C. Chang, "Accumulation of stevioside and
rebaudioside A in callus cultures of Stevia rebaudiana Bertoni", Bot.
Bull. Acad. Sin. vol. 24, pp. 115-119, 1983.
[5] P. Jain, S. Kaxhhwaha, S. L. Kothari, "Improved micropropagation
protocol and enhancement in biomass and chlorophyll content in Stevia
rebaudiana (Bert.) Bertoni by using high copper levels in the culture
medium", Scientia Horticulturae, vol. 119, pp.315-319, 2009.
[6] B. Janarthanam, M. Gopalakrishnan, S G. Lakshmi, T. Sekar, "Plant
regeneration from leaf derived callus of Stevia rebaudiana Bertoni",
Plant Tissue Culture and Biotechnology, vol.19, no. 2, pp. 133-141,
2009.
[7] A.D. Kinghorn, D.D. Soejarto, "Sweetening agents of plant origin".
Crit. Rev. Plant Sci. 4, pp. 79-120, 1986.
[8] F.A. Krens, Jamar. D, "The role of explant source and culture
conditions on callus induction and shoot regeneration in sugarbeet
(Beta vulgaris L.)", J. Plant Physiol, vol. 134, pp. 651-655, 1989.
[9] A.G. Lyakhovkin, T.D. Long, D.A. Titov, M.P. Anh, "In: Cultivation
and utilization of Stevia (Stevia rebaudiana Bertoni)", Agricultural
Publishing House, Hanoi, pp. 1-44, 1993.
[10] R.M. Patel, R. R. Shah, "Regeneration of Stevia Plant through Callus
Culture", Indian Journal of Pharmaceutical Sciences, pp. 46-50, vol.2,
Jan-Feb. 2009.
[11] L. Sivaram, U. Mukundan, "In vitro culture studies on Stevia
rebaudiana", In Vitro Cell. Dev. Biol.-Plant, vol. 39, pp 520-523,
September-October. 2003.
[12] S.M. Swanson, G.B. Mahady, C.W.W. Beecher, "Stevioside
biosynthesis by callus, root, shoot and rooted-shoot cultures in vitro".
Plant Cell Tissue Organ Cult. Vol. 28, pp. 151-157 1992.
[13] M.B. Tadhni, V.H. Patel, R. Subhash, "In vitro antioxidant activities of
Stevia rebaudiana leaves and callus". Journal of food composition and
Analysis , vol.20, pp. 323-329, 2007.
[14] A.S. Taware, D. S. Mukadam, A. M. Chavan, S.D. Taware,
"Comparative study of in-vitro and in-vivo grown plants and callus of
Stevia rebaudiana (Bertoni)", International Journal of integrative
Biology, vol.9, no. 1, pp 10-15, 2010.
[15] MS. Uddin, M. S. Chowdhary, H. Khan, "In vitro propagation of Stevia
rebaudiana Bert in Bangladesh", African Journal of Biotechnology,
vol. 5, no.13, pp. 1238-1240, July. 2006.
[16] T. Yamazaki, H.E. Flores, "Examination of steviol glycoside
production by hair root and shoot cultures of Stevia rebaudiana". J.
Nat. Prod, vol 54, pp. 986-992, 1991.
@article{"International Journal of Biological, Life and Agricultural Sciences:50458", author = "Pratibha Gupta and Satyawati Sharma and Sanjay Saxena", title = "Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production", abstract = "Stevia rebaudiana Bertoni (natural sweetener) belongs
to Asteraceae family and can be used as substitute of artificial
sweeteners for diabetic patients. Conventionally, it is cultivated by
seeds or stem cutting, but seed viability rate is poor. A protocol for
callus induction and multiplication was developed to produce large
no. of calli in short period. Surface sterilized nodal, leaf and root
explants were cultured on Murashige and Skoog (MS) medium with
different concentrations of plant hormone like, IBA, kinetin, NAA,
2,4-D, and NAA in combination with 2,4-D. 100% callusing was
observed from leaf explants cultured on combination of NAA and
2,4-D after three weeks while with 2,4-D, only 10% callusing was
observed. Calli obtained from leaf and root explants were shiny green
while with nodal explants it was hard and brown. The present
findings deal with induction of callusing in Stevia to achieve the
rapid callus multiplication for study of steviol glycosides in callus
culture.", keywords = "2,4-D, Callusing, NAA, Stevia, Steviol glycosides", volume = "4", number = "12", pages = "870-5", }
