Effects of Explant Type and Plant Growth Regulator Combinations on In Vitro Direct and Indirect Regeneration of Stevia rebaudiana

Document Type : Research Article

Authors

1 Assistant Professor, Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

2 MSc Graduated Student, Department of Plant Breeding, Faculty of Agriculture, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 MSc Graduated Student, Department of Horticulture, Faculty of Agriculture, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

4 BSc Graduated Student, Department of Horticulture, Faculty of Agriculture, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

5 MSc Graduated Student, Department of Breeding, Faculty of Agriculture, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

Abstract

Stevia rebaudiana is a medicinal plant belonging to Asteraceae family. This experiment was carried out to optimize the in vitro callus induction and plant regeneration of Stevia rebaudiana. The explants used in this study were leaf, shoot tip, nodal segments with two leaves and nodal segment without leaves derived from in vitro grown plantlets. The explants were transferred to PGR-free MS medium or supplemented with BA (0.1, 0.5 and 2mgl-1) solely or in combination with 0.5mgl-1 IBA and TDZ (0.1, 0.5 and 2mgl-1) solely or in combination with 0.5mgl-1 IBA. The highest callus percentage and callus volume induced with culturing leaf explant on MS medium containing 0.1 mgl-1 TDZ. The nodal segment without leaves cultured on MS medium containing 2mgl-1 BA + 0.5mgl-1 IBA produced the highest regenerated shoot number. The highest and lowest shoot length was observed in shoot tip cultured on MS medium containing 0.1mgl-1 BA + 0.5mgl-1 IBA and MS medium containing 2mgl-1 BA + 0.5mgl-1 IBA, respectively. Indirect shoot regeneration was observed only in leaf explant cultured on MS medium containing 0.5mgl-1 TDZ + 0.5mgl-1 IBA. The highest percentage of root induction was observed with culturing shoots on MS medium containing 0.1mgl-1 IBA.

Keywords

Main Subjects


 Abdul Razak, U. N. A., Ong, Ch. 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: 23-28.
Ahmed, S. R., Howlader, M. M. S., Sutradhar, P. and Yasmin, S. 2016. An efficient protocol for in vitro regeneration of Stevia rebaudiana. Asian Journal of Medical and Biological Research, 2 (1): 95-106.
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: 1263-1266.
Curry, L. L. and Ashley, R. 2008. Subchronic toxicity of rebaudioside A. Cargill Incorporated. Journal Process Biochemistry, 83: 115-117.
Das, A., Gantait, S. and Mandal, N. 2011. Micropropagation of an elite medicinal plant: Stevia rebaudiana Bert. International Journal of Agricultural Research, 6: 40-48.
George, E. F. and Sherrington, P. D. 1984. Plant propagation by tissue culture. Exegetics Ltd., Eversley, Basingstoke.
Jaroslov, P., Brabora, H. and Tuulia, H. 2007. Characterisation of steviol rebaudiana by Choprehensive tow- dimensional liquid chromatography time of flight mass spectrometry. Journal of Chromatography, 1150 (1-2): 85-92.
Jitendra, M., Monika, S., Ratan, S. D., Priyanka, G., Priyanka, S. and Kiran, D. J. 2012. Micropropagation of an anti diabetic plant - Stevia rebaudiana Bertoni, (Natural Sweetener) in Hadoti region of South-East Rajasthan, India, ISCAJournal of Biological Sciences, 1 (3): 37-42.
Kim, N. C. and Kinghorn, A. D. 2002. Highly sweet compounds of plant origin. Archives of Pharmacal Research, 25: 725-746.
Laribi, B., Roatbi, N., Kouki, K. and Bettaieb, T. 2012. In vitro propagation of stevia rebaudiana (Bert.) - a non caloric sweetener and antidiabetic medicinal plant. International Journal of Medicinal and Aromatic Plants, 2: 333-339.
Mizutani, K. O. and Tanaka, A. 2002. Use of Stevia rebaudiana sweeteners in Japan. In: Stevia, the Genus Stevia. Medicinal and aromatic plants-industrial profiles. Taylor and Francis: London and New York, 19: 178-195.
Mohamed, R. A. and Alhady, A. 2011. Micropropagation of Stevia rebaudiana Bertoni. A new sweetening crop in Egypt. Global Journal of Biotechnology and Biochemistry, 6 (4): 178-182.
Patel, R. M. and Shah, R. R. 2009. Regeneration of stevia plant through callus culture. Indian Journal of Pharmaceutical Sciences, 71: 46-50.
Rafiq, M., Dahot, M. U., Muhamamd, Sh., Naqvi, H. A. and Qarshi, I. A. 2007. In vitro clonal propagation and biochemical analysis of field established Stevia rebaudiana bertoni. Pakistan Journal of Botany, 39 (7): 2467-2474.
Sharma, N., Gauchan, D. P., Dhakal, A., Luitel, A., Shakya, S. and Shakya, R. 2015. Establishment of regenerative callus, cell suspension system and molecular characterization of stevia rebaudiana Bertoni for the production of stevioside in in vitro. International Journal for Research in Applied Science and Engineering Technology (IJRASET), 3: 133-144.
Soejarto, D. D., Compadre, C., Medon, P. J., Kamath, S. K. and Kinghorn, A. D. 1983. Potential sweetening agents of plant origin. II. Field research for sweet tasting Stevia species. Economic Botany, 37: 71-79.