STANDARDIZATION OF SURFACE STERILIZATION PROTOCOL OF FIELD GROWN Stevia rebaudiana PRIOR TO IN VITRO CLONAL PROPAGATION
DOI:
https://doi.org/10.11113/jt.v77.6722Keywords:
In vitro propagation, Surface sterilization, Stevia rebaudiana, NaOCl, HgCl2Abstract
The high demand of Stevia rebaudiana, commonly known as Stevia encourages people for commercial-scale cultivation. In vitro clonal propagation is a potential alternative for conventional propagation due to its advantages. However, microbial contamination is a constant problem for in vitro clonal propagation of field grown plant. Therefore, the aim of this study was to establish an effective surface sterilization protocol for Stevia prior to in vitro propagation. Two disinfecting agents, NaOCl and HgCl2 with different concentrations and various exposure times were tested for surface sterilization of shoot tips and nodal explants. Results show that treatment of explants with NaOCl was more effective compared to HgCl2 and 5% NaOCl for 10 minutes with 0.2% carbendazim (fungicide) pre-treatment showed least contamination and highest survival rate. Fungal contamination was observed to be a main problem in this study and it occurred within 2-3 days of inoculation, leading to the death of the explants.
References
Shivaprasad, H. N. 2008. Expectations of Natural/Herbal Products Industry. In: International Conclave on Medicinal Plants for ASEAN and BIMSTEC Countries. 11-13 December, 2008 Jubilee Hall RIMS Campus, Lamphelpat, Imphal, Manipur, India. 78-82.
Hossain, M. S., Urbi, Z., Sule, A. and Rahman, K. 2014. Andrographis Paniculata (Burm. F.) Wall. Ex Nees: A Review of Ethnobotany, Phytochemistry, and Pharmacology. The Scientific World Journal. 2014: 28.
Bagozzi, D. 2003. Traditional Medicine, Fact Sheet N°134. World Healt Organization. Retrieved on 29 July 2015 from [http://www.who.int/mediacentre/factsheets/2003/fs134/en/print.html].
MPHPBPC. 2015. Traditional Medicine & Herbal Medicine. Ministry of Commerce, Bangladesh. Retrieved on 29 July from [http://bpc.org.bd/mphpbpc_sector_profile.php].
Duke, J. A. and duCellier, J. L. 1993. Crc Handbook of Alternative Cash Crops. CRC Press, Taylor & Francis Group.
Soejarto, D. D., Kinghorn, A. D. and Farnsworth, N. R. 1982. Potential Sweetening Agents of Plant Origin. Iii. Organoleptic Evaluation of Stevia Leaf Herbarium Samples for Sweetness. Journal of Natural Products. 45: 590-599.
Stones, M. 2011. Stevia Wins Final Eu Approval. foodmanufacture.co.uk. Retrieved on 16 September 2014 from [http://www.foodmanufacture.co.uk/Ingredients/Stevia-wins-final-EU-approval].
Lucas, L. 2011. Brussels Backs Stevia Sweetener. Financial Times. Retrieved on 16 September 2014 from [http://www.ft.com/cms/s/0/f1e157e0-0ec1-11e1-b83c-00144feabdc0.html].
Carakostas, M., Curry, L., Boileau, A. and Brusick, D. 2008. Overview: The History, Technical Function and Safety of Rebaudioside a, a Naturally Occurring Steviol Glycoside, for Use in Food and Beverages. Food and Chemical Toxicology. 46: S1-S10.
Lemus-Mondaca, R., Vega-Gálvez, A., Zura-Bravo, L. and Ah-Hen, K. 2012. Stevia Rebaudiana Bertoni, Source of a High-Potency Natural Sweetener: A Comprehensive Review on the Biochemical, Nutritional and Functional Aspects. Food Chemistry. 132: 1121-1132.
Chong, J. K. 2013. 4th National Diabetes Conference 2013. 14-15 June, 2013. Hotel Istana, Kuala Lumpur, Malaysia. themalaysianinsider.com. Retrieved on 11 July 2013 from [http://www.themalaysianinsider.com/malaysia/article/number-of-diabetics-in-malaysia-alarming-says-council/].
Brandle, J., Starratt, A. and Gijzen, M. 1998. Stevia Rebaudiana: Its Agricultural, Biological, and Chemical Properties. Canadian Journal of Plant Science. 78: 527-536.
Abudula, R., Jeppesen, P. B., Rolfsen, S. E. D., Xiao, J. and Hermansen, K. 2004. Rebaudioside a Potently Stimulates Insulin Secretion from Isolated Mouse Islets: Studies on the Dose-, Glucose-, and Calcium-Dependency. Metabolism. 53: 1378-1381.
Jeppesen, P. B., Gregersen, S., Rolfsen, S., Jepsen, M., Colombo, M., Agger, A., Xiao, J., Kruhøffer, M., Ørntoft, T. and Hermansen, K. 2003. Antihyperglycemic and Blood Pressure-Reducing Effects of Stevioside in the Diabetic Goto-Kakizaki Rat. Metabolism. 52: 372-378.
Odutayo, O., Amusa, N., Okutade, O. and Ogunsanwo, Y. 2007. Sources of Microbial Contamination in Tissue Culture Laboratories in Southwestern Nigeria. African Journal of Agricultural Research. 2: 067-072.
Murashige, T. and Skoog, F. 1962. A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum. 15: 473-497.
Shatnawi, M., Shibli, R., Abu-Romman, S., Al-Mazra’awi, M., Al Ajlouni, Z., Shatanawi, W. and Odeh, W. 2011. Clonal Propagation and Cryogenic Storage of the Medicinal Plant Stevia Rebaudiana. Spanish Journal of Agricultural Research. 9: 213-220.
Abd Alhady, M. R. A. 2011. Micropropagation of Stevia Rebaudiana Bertoni—a New Sweetening Crop in Egypt. Global J. Biotechnol. Biochem. 6: 178-182.
Jain, P., Kachhwaha, S. and Kothari, S. 2012. Optimization of Micronutrients for the Improvement of in Vitro Plant Regeneration of Stevia Rebaudiana (Bert.) Bertoni. Indian Journal of Biotechnology. 11: 486-490.
Hassanen, S. A. and Khalil, R. M. 2013. Biotechnological Studies for Improving of Stevia (Stevia Rebaudiana Bertoni) in Vitro Plantlets. Middle-East Journal of Scientific Research. 14: 93-106.
Thiyagarajan, M. and Venkatachalam, P. 2012. Large Scale in Vitro Propagation of Stevia Rebaudiana (Bert) for Commercial Application: Pharmaceutically Important and Antidiabetic Medicinal Herb. Industrial Crops and Products. 37: 111-117.
Anbazhagan, M., Kalpana, M., Rajendran, R., Natarajan, V. and Dhanavel, D. 2010. In Vitro Production of Stevia Rebaudiana Bertoni. Emirates Journal of Food and Agriculture. 22: 216.
Laribi, B., Rouatbi, 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.
Pierik, R. L. M. 1997. In Vitro Culture of Higher Plants. Springer Science & Business Media. 89-94.
Himabindu, K., Priya, M. S., Reddy, D. M., Sudhakar, P., Srinivasulu, Y., Reddisekhar, M., Latha, P. and Reddy, B. R. K. 2012. Studies on the Effect of Various Sterilants and Culture Conditions on in-Vitro Seed Germination in Tomato (Solanum Lycopersicum). International Journal of Applied Biology and Pharmaceutical Technology. 3: 476-480.
Nakagawara, S., Goto, T., Nara, M., Ozawa, Y., Hotta, K. and Arata, Y. 1998. Spectroscopic Characterization and the pH Dependence of Bactericidal Activity of the Aqueous Chlorine Solution. Analytical Sciences. 14: 691-698.
Badoni, A. and Chauhan, J. 2010. In Vitro Sterilization Protocol for Micropropagation of Solanum Tuberosum Cv ‘Kufri Himalini’. Acedemia Arena. 2: 24-28.
Tiwari, S., Arya, A. and Kumar, S. 2012. Standardizing Sterilization Protocol and Establishment of Callus Culture of Sugarcane for Enhanced Plant Regeneration in Vitro. Research Journal of Botany. 7: 1-7.
Patel, D. R., Subash, N., Patel, A. M. and Patel, S. A. 1997. Mass in Vitro Multiplication of Kankoda (Momordica Dioica L.). Plant Struct. Morph. 15: 105-109.
Reddy, S. P., Gopal, R. G. and Sita, L. G. 1998. In Vitro Multiplication of Gymnema Sylvestre R. Br.–an Important Medicinal Plant. Current Science. 75: 843-845.
Kataky, A. and Handique, P. 2010. Micropropagation and Screening of Antioxidant Potential of Andrographis Paniculata (Burm. F) Nees. Journal of Hill Agriculture. 1: 13-18.
Downloads
Published
Issue
Section
License
Copyright of articles that appear in Jurnal Teknologi belongs exclusively to Penerbit Universiti Teknologi Malaysia (Penerbit UTM Press). This copyright covers the rights to reproduce the article, including reprints, electronic reproductions, or any other reproductions of similar nature.
