Micropropagation of Stevia by Tissue Cultures in a Temporary Immersion System
Abstract
The aim of this study was to evaluate the temporary immersion bioreactor in multiplication of Stevia. The first factor was the immersion frequencies. Nodal explants were fed with liquid MS medium for 3 minutes each 3, 4, 6, 8 and 12 hours. Cultivate for 4 weeks. It was found that the 3 hours immersion frequency had the highest average dry weight of 40.8 mg and the highest average shoot number was 3.6 shoots per explant with statistically significant difference at 0.05. To study effects of immersion time in TIG. Cultures were immersed in medium for 1, 3, 5, 7 and 10 minutes, every 3 hours in TIG for 4 weeks. It was found that the 3 minutes immersion time showed the highest average dry weight and the average number of shoots with significantly difference higher than the other treatments, 40.5 mg and 3.6 shoots per explant respectively. To compared of different in vitro micropropagation methods of Stevia. In vitro explants were cultured in TIG by 3 hours immersion frequency with a 3 minutes immersion, liquid medium at 110 rpm speed shaker and agar medium in light condition 16 hours per day at 25°C cultured for 4 weeks. The results show that the cultured in the immersion system showed the highest average dry weight 40.5 mg which more than cultured in liquid medium and agar medium with statistically significant difference at 0.05. While, cultivation in TIG and agar medium provides a nonsignificant difference in the average number of 3.6 and 3.1 shoots per explant and liquid medium produced the lowest average number of shoots with statistically significant difference at 0.05. Therefore, the cultivation of stevia explant in TIG which is an alternative to application for the production of stevia at the industrial level. Keywords : stevia, propagation, plant tissue cultures, temporary immersion systemReferences
Alexander V. & Esyanti R. (2016). Stimulation of stevioside accumulation on Stevia rebaudiana (Bertoni) shoot culture induced with red LED light in TIS RITA® bioreactor system. World Academy of Science, Engineering and Technology International Journal of Biotechnology and Bioengineering, 10(12), 803-808.
Bhatia S. & Sharma K. (2015). Technical glitches in micropropagation. In Modern Applications of Plant Biotechnology in Pharmaceutical Sciences. (pp. 393-404). Elsevier Incorporated.
Chotikadachanarong K. & Dheeranupattana S. (2013). Micropropagation and acclimatization of Stevia rebaudiana Bertoni. Pakistan Journal of Biological Sciences, 16(7), 887-890.
Etienne, H., Lartaud, M., Michaux-Ferriere, N., Carron, M. P., Berthouly, M. & Teisson, C. (1997). Improvement of somatic embryogenesis in Hevea brasiliensis using the temporary immersion technique. In Vitro Cellular and Developmental Biology of Plant, 33, 81-87.
Kongbangkerd, A., Wongsa, T. & Wannachart, S. (2012). Factors affecting growth and development of in vitro young shoot culture of Dendrobium ochreatum Lindl. in a temporary immersion system. Thai Journal of Botany, 4(Special Issue), 159-168. (In Thai)
Madan, S., Ahmad, S., Singh, G. N., Kohli, K., Kumar, Y., Singh, R. & Garg, M. (2010) Stevia rebaudiana (Bert.) Bertoni-A review. Indian Journal of Natural Products and Resources, 1, 267-286.
Miyazaki, Y. & Wantenabe, H. (1974). Studies on the cultivation of Stevia rebaudiana Bertoni; on the propagation of the plant (English abstr.). Japanese Journal of Tropical Agriculture, 17, 154 – 157.
Mongkolsook, Y. & Pongchan, W. (2000). Gloxinia (Sinningia speciosa) micropropagation by using temporary immersion technique. In Proceedings of the 38th Kasetsart University Annual Conference: Plants, Agricultural Extension and Communication. (pp:387-390). Kasetsart University. (in Thai)
Ramirez-Mosqueda. M. A., Iglesias-Andreu, L. G., Ramirez-Madero, G. & Hernandez-Rincon, E. U. (2016). Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. South African Journal of Botany, 106, 238-243.
Rangsayatorn, N. (2014). Micropropagation of Dendrobium draconis Rchb.f. by using temporary immersion system. In Proceedings of 51st Kasetsart University Annual Conference: Plants. (pp:312-318).
Kasetsart University. (in Thai)
Sumkaew, R., Pankaew, Y., Puangchit, L., Siripatanadilok, S. & Kokkatiem, S. (2010) In vitro seedling of Eucalyptus camaldulensis Dehnh. using temporary immersion system with twin flasks. In Proceedings of 48st Kasetsart University Annual Conference: Plants. (pp:514-522). Kasetsart University. (in Thai)
Venutolo, S. A. & Aguilar, T. S. (2015). Mass micropropagation of Stevia rebaudiana Bertoni in temporary immersion systems. Cultivos Tropicales, 36(3), 50-57.
Vives, K., Andujar, I., Lorenzo, J. C. & Concepcion, O. (2017). Comparison of different in vitro micropropagation methods of Stevia rebaudiana B. including temporary immersion bioreactor (BIT®). Plant Cell, Tissue and Organ Culture, 131, 195-199.
Bhatia S. & Sharma K. (2015). Technical glitches in micropropagation. In Modern Applications of Plant Biotechnology in Pharmaceutical Sciences. (pp. 393-404). Elsevier Incorporated.
Chotikadachanarong K. & Dheeranupattana S. (2013). Micropropagation and acclimatization of Stevia rebaudiana Bertoni. Pakistan Journal of Biological Sciences, 16(7), 887-890.
Etienne, H., Lartaud, M., Michaux-Ferriere, N., Carron, M. P., Berthouly, M. & Teisson, C. (1997). Improvement of somatic embryogenesis in Hevea brasiliensis using the temporary immersion technique. In Vitro Cellular and Developmental Biology of Plant, 33, 81-87.
Kongbangkerd, A., Wongsa, T. & Wannachart, S. (2012). Factors affecting growth and development of in vitro young shoot culture of Dendrobium ochreatum Lindl. in a temporary immersion system. Thai Journal of Botany, 4(Special Issue), 159-168. (In Thai)
Madan, S., Ahmad, S., Singh, G. N., Kohli, K., Kumar, Y., Singh, R. & Garg, M. (2010) Stevia rebaudiana (Bert.) Bertoni-A review. Indian Journal of Natural Products and Resources, 1, 267-286.
Miyazaki, Y. & Wantenabe, H. (1974). Studies on the cultivation of Stevia rebaudiana Bertoni; on the propagation of the plant (English abstr.). Japanese Journal of Tropical Agriculture, 17, 154 – 157.
Mongkolsook, Y. & Pongchan, W. (2000). Gloxinia (Sinningia speciosa) micropropagation by using temporary immersion technique. In Proceedings of the 38th Kasetsart University Annual Conference: Plants, Agricultural Extension and Communication. (pp:387-390). Kasetsart University. (in Thai)
Ramirez-Mosqueda. M. A., Iglesias-Andreu, L. G., Ramirez-Madero, G. & Hernandez-Rincon, E. U. (2016). Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. South African Journal of Botany, 106, 238-243.
Rangsayatorn, N. (2014). Micropropagation of Dendrobium draconis Rchb.f. by using temporary immersion system. In Proceedings of 51st Kasetsart University Annual Conference: Plants. (pp:312-318).
Kasetsart University. (in Thai)
Sumkaew, R., Pankaew, Y., Puangchit, L., Siripatanadilok, S. & Kokkatiem, S. (2010) In vitro seedling of Eucalyptus camaldulensis Dehnh. using temporary immersion system with twin flasks. In Proceedings of 48st Kasetsart University Annual Conference: Plants. (pp:514-522). Kasetsart University. (in Thai)
Venutolo, S. A. & Aguilar, T. S. (2015). Mass micropropagation of Stevia rebaudiana Bertoni in temporary immersion systems. Cultivos Tropicales, 36(3), 50-57.
Vives, K., Andujar, I., Lorenzo, J. C. & Concepcion, O. (2017). Comparison of different in vitro micropropagation methods of Stevia rebaudiana B. including temporary immersion bioreactor (BIT®). Plant Cell, Tissue and Organ Culture, 131, 195-199.
Downloads
Published
2020-01-15
Issue
Section
Research Article
