Induction of Callus and Ploidy Level Inspection from Tissue Culture of Sundew (Drosera indica L.) in Sterile Condition
Abstract
The purpose of this study was to induce callus and determine ploidy level from tissue culture of Drosera indica L. in sterile conditions. The leaf explants of Sundew were cultured on ½ MS (Murashige and Skoog, 1962) media supplemented with BA and NAA at different concentrations for 4 weeks. From the results, it was found that leaf explants cultured on ½ MS media containing 0.5 mg / l BA with NAA 0.1 mg / l could induce callus well and gave the highest callus induction percentage and callus fresh weight. On the other hand, leaf explants cultured on MS media supplemented with 1.0 mg / l BA in combination with 0.1 mg / l NAA had the highest number of shoot induction from callus. Then, the callus was cultured on ½ MS media without plant growth regulators for 4 weeks to induce new shoot. After that, cut the shoot to get the node explant size 2 ± 0.2 cm, then dipped in the ½ MS liquid media with the addition of 0, 0.1, 0.5, and 1.0 mg / l colchicine at period of 24, 48 and 72 hours. Then node explants were cultured on ½ MS solid media for 4 weeks. It was found that node explants immersing in liquid MS media without adding colchicine had the highest survival percentage. Furthermore, it could be seen that the survival percentage decreased as the concentration of colchicine increased as well as the immersion period increased. When analyzing ploidy levels with the flow cytometry technique, it was found that there were a number of chromosomes in the diploid. Then, the seedlings from tissue culture were transplanted in peat moss together with the sphagnum moss in the ratio of 1 : 2 for 4 weeks. The result showed that seedlings had the highest survival percentage. Keywords : callus, colchicine, flow cytometry, ploidy, Drosera indica L.References
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Balla, N., Taychasinpitak, T. & Thanananta, T. (2007). Induce Polyploid Ornamatal Sweet Potato in Vitro by Colchicine. Thai Science and Technology Journal, 25, 261–266. (in Thai)
Buddharaksa, P. (2013). Plant Tissue Culture. Chiang mai: Nopburee Press Company Printing Design in
Chiang mai. (in Thai)
Charoenwattana, P. (2014). Invitro Cultures and Colchicin – Induced Tetraploidy of Sundew (Drosera spatulate Labill.). International Journal of Enviromental and Rural Development, 5 (2), 80-85.
Chuntaratin, P. (2013). Collection and Production of Carnivorous Plants in the Genus Drosera by Plant Tissue Culture Technique. Bangkok: Agricultural Technology Program, Bangkok Thonburi University. Retrieved July 1, 2018, from http://qa.bkkthon.ac.th/qa/qa56/132144099.pdf. (in Thai)
Chiang, C.P. (2010). Growing Carnivorous Plants in the Topics. Singapore : Celestial.
Dermen, H. (1940). Colchicine polyploidy and technique. The Botanical Review, 6,(11), 599-635.
Dhar, U. & Joshi, M. (2005). Efficient plant regeneration protocol through callus for Saussurea obvallata (DC.) Edgew. (Asteraceae): effect of explant type, age and plant growth regulators. Plant Cell Report, 24, 195-200.
Didry, N., Dubreuil, L., Trotin, F. & Pinkas, M. (1998). Antimicrobial activity of aerial parts of Drosera peltata Snith on oral bacteria. Journal of Ethnopharmacology, 60, 91-96.
Hanson, L., Mcmahon, K., Johnson, M. & Bennett, M. (2001). First nuclear DNA C – values for 25 angiosperm families. Annals of Botany, 87, 251-258.
Jala, A. & Paitoon, S. (2013). Micropropagation of Dionaea muscipula by Tissue culture. Thai Journal of Science and Technology, 2(2), 134-139. (in Thai)
Jayaram, K. & Prasad, M. (2007). Rapid in vitro multiplication of Drosera indica L.: a vulnerable, medicinally important insectivorous plant. Plant Biotechnology Reports, 1(2), 79-84.
Juengwatanatrakul, T., Sakamoto, S., Tanaka, H. & Putalun, W. (2011). Eliciation effect on production of plumbagin in vitro of Drosera indica L.. Journal of Medicinal Plants Research, 5(19), 4949-4953.
Kauth, P.J., Vendrame, W.A. & Kane, M.E. (2006). In vitro seed culture and seedling development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture, 85, 91–102.
Kim, K.S. & Jang, G.W. (2004). Micropropagation of Drosera peltata , a tuberous sundew, by shoot tip culture. Plant Cell, Tissue and Organ Culture, 77, 211-214.
Kongbangkerd, A., Homchan, S. & Promthep, K. (2011). Effect of plant growth regulators and plant growth retardants on growth and development on in vitro leaf culture of Drosera communis St.Hil. NU Science Journal, 8(1), 87 - 100. (in Thai)
Lee, L. (2008). Carnivorous plants: new ornamentals. Chronica Horticulturae, 48(4), 11-14.
Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.
Neera, S., Soonthorn, W., Mongkolpun, P. & Hongpukdee, P. (2014). Effect of cytokinins and coconut water on in vitro shoot induction of Kaentawan (Helianthus tuberous L.). khon kaen agriculture journal, 42(3),
328-334. (in Thai)
Pavla, Z., Barbora, S., Ales, H. & Eloy, C. (2013). In vitro Induced Mitotic Polyploidy in Drosera capensis L..
Agricultura tropica et subtropica, 46(4), 107-110.
Petchang, R. (2013). Propagation of some local plants by tissue culture in Tumbon Nanokkok, Lablae, Uttaradit.
Naresuan University Journal: Science and Technology, 19(3), 35-42. (in thai)
Pimsuwan, S. (2012) Research report Effect of Various Type of Growing Media on Growth of Venus Flytrap, (Dionaea muscipula), Faculty of Agricultural Technology Rajamangala University of Technology Thanyaburi. 64 pp. (in thai)
Promchan, T., Ramasoot, S. & Bunwest, P. (2016) Effects of Activated Charcoal and Growing Media on Seedling
Growth and Survival Rate of Aeridesodorata. Wichcha Journal, 35(2), 53-61. (in Thai)
Samala, S., Jansedom, H. & Saehan, O. (2015). Effect of Colchicin on Morphological Characteristics of In Vitro Drosera burmannii. Songklanakarin Journal of plant Science, 2(1), 24 - 28. (in Thai)
Taweerodjanakarn, S., Kongton, K., Thongyai. K. & Samala, S. (2018). Invitro Micropropagation of Drosera burmanni Vahl. Songklanakarin Journal of Plant Science, 5(3). 18-26. (in Thai)
Thengane, S. R., Joshi, M. S., Khuspe, S. S. & Mascarenhas, A. F. (1994). Anther culture in Helianthus annuus L., influence of genotype and culture conditions on embryo induction and plant regeneration. Plant cell reports, 13(3-4), 222-226.
Tungsunan, T., Chotikadachanarong, S. & Pichai, K. (2015). Invitro propagation of stevia (stevia rebaudiana Bertoni.). Journal of Thai Interdisciplinary Resarch, 10(1), 8-16. (in Thai)
Yusoh, A. & Te-chato, S. (2014). Effect of Sucrose and Dicamba on Callus Induction from Mature Seeds and Young Leaves Culture of Hom Kra-Dang-Nga Rice (Indica). Songklanakarin Journal of Plant Science, 1(3), 28-31. (in thai)
Balla, N., Taychasinpitak, T. & Thanananta, T. (2007). Induce Polyploid Ornamatal Sweet Potato in Vitro by Colchicine. Thai Science and Technology Journal, 25, 261–266. (in Thai)
Buddharaksa, P. (2013). Plant Tissue Culture. Chiang mai: Nopburee Press Company Printing Design in
Chiang mai. (in Thai)
Charoenwattana, P. (2014). Invitro Cultures and Colchicin – Induced Tetraploidy of Sundew (Drosera spatulate Labill.). International Journal of Enviromental and Rural Development, 5 (2), 80-85.
Chuntaratin, P. (2013). Collection and Production of Carnivorous Plants in the Genus Drosera by Plant Tissue Culture Technique. Bangkok: Agricultural Technology Program, Bangkok Thonburi University. Retrieved July 1, 2018, from http://qa.bkkthon.ac.th/qa/qa56/132144099.pdf. (in Thai)
Chiang, C.P. (2010). Growing Carnivorous Plants in the Topics. Singapore : Celestial.
Dermen, H. (1940). Colchicine polyploidy and technique. The Botanical Review, 6,(11), 599-635.
Dhar, U. & Joshi, M. (2005). Efficient plant regeneration protocol through callus for Saussurea obvallata (DC.) Edgew. (Asteraceae): effect of explant type, age and plant growth regulators. Plant Cell Report, 24, 195-200.
Didry, N., Dubreuil, L., Trotin, F. & Pinkas, M. (1998). Antimicrobial activity of aerial parts of Drosera peltata Snith on oral bacteria. Journal of Ethnopharmacology, 60, 91-96.
Hanson, L., Mcmahon, K., Johnson, M. & Bennett, M. (2001). First nuclear DNA C – values for 25 angiosperm families. Annals of Botany, 87, 251-258.
Jala, A. & Paitoon, S. (2013). Micropropagation of Dionaea muscipula by Tissue culture. Thai Journal of Science and Technology, 2(2), 134-139. (in Thai)
Jayaram, K. & Prasad, M. (2007). Rapid in vitro multiplication of Drosera indica L.: a vulnerable, medicinally important insectivorous plant. Plant Biotechnology Reports, 1(2), 79-84.
Juengwatanatrakul, T., Sakamoto, S., Tanaka, H. & Putalun, W. (2011). Eliciation effect on production of plumbagin in vitro of Drosera indica L.. Journal of Medicinal Plants Research, 5(19), 4949-4953.
Kauth, P.J., Vendrame, W.A. & Kane, M.E. (2006). In vitro seed culture and seedling development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture, 85, 91–102.
Kim, K.S. & Jang, G.W. (2004). Micropropagation of Drosera peltata , a tuberous sundew, by shoot tip culture. Plant Cell, Tissue and Organ Culture, 77, 211-214.
Kongbangkerd, A., Homchan, S. & Promthep, K. (2011). Effect of plant growth regulators and plant growth retardants on growth and development on in vitro leaf culture of Drosera communis St.Hil. NU Science Journal, 8(1), 87 - 100. (in Thai)
Lee, L. (2008). Carnivorous plants: new ornamentals. Chronica Horticulturae, 48(4), 11-14.
Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.
Neera, S., Soonthorn, W., Mongkolpun, P. & Hongpukdee, P. (2014). Effect of cytokinins and coconut water on in vitro shoot induction of Kaentawan (Helianthus tuberous L.). khon kaen agriculture journal, 42(3),
328-334. (in Thai)
Pavla, Z., Barbora, S., Ales, H. & Eloy, C. (2013). In vitro Induced Mitotic Polyploidy in Drosera capensis L..
Agricultura tropica et subtropica, 46(4), 107-110.
Petchang, R. (2013). Propagation of some local plants by tissue culture in Tumbon Nanokkok, Lablae, Uttaradit.
Naresuan University Journal: Science and Technology, 19(3), 35-42. (in thai)
Pimsuwan, S. (2012) Research report Effect of Various Type of Growing Media on Growth of Venus Flytrap, (Dionaea muscipula), Faculty of Agricultural Technology Rajamangala University of Technology Thanyaburi. 64 pp. (in thai)
Promchan, T., Ramasoot, S. & Bunwest, P. (2016) Effects of Activated Charcoal and Growing Media on Seedling
Growth and Survival Rate of Aeridesodorata. Wichcha Journal, 35(2), 53-61. (in Thai)
Samala, S., Jansedom, H. & Saehan, O. (2015). Effect of Colchicin on Morphological Characteristics of In Vitro Drosera burmannii. Songklanakarin Journal of plant Science, 2(1), 24 - 28. (in Thai)
Taweerodjanakarn, S., Kongton, K., Thongyai. K. & Samala, S. (2018). Invitro Micropropagation of Drosera burmanni Vahl. Songklanakarin Journal of Plant Science, 5(3). 18-26. (in Thai)
Thengane, S. R., Joshi, M. S., Khuspe, S. S. & Mascarenhas, A. F. (1994). Anther culture in Helianthus annuus L., influence of genotype and culture conditions on embryo induction and plant regeneration. Plant cell reports, 13(3-4), 222-226.
Tungsunan, T., Chotikadachanarong, S. & Pichai, K. (2015). Invitro propagation of stevia (stevia rebaudiana Bertoni.). Journal of Thai Interdisciplinary Resarch, 10(1), 8-16. (in Thai)
Yusoh, A. & Te-chato, S. (2014). Effect of Sucrose and Dicamba on Callus Induction from Mature Seeds and Young Leaves Culture of Hom Kra-Dang-Nga Rice (Indica). Songklanakarin Journal of Plant Science, 1(3), 28-31. (in thai)
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2019-09-16
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