Effects of Culture Medium on In Vitro Growth and Development of Dendrobium garrettii Seidenf., A Rare Orchid of Thailand
Dendrobium garrettii Seidenf. is a miniature orchid, which is native and found only in Thailand. This species consumes time for growth, and is confronting with the problem of smuggling out of the natural habitat leading to a dramatic decrease in its population. Therefore, its status is an endangered species. To conserve and prevent extinction, this research established the suitable medium for ex situ conservation of D. garrettii based on plant tissue culture. Two steps of study were done by clarifying the appropriate strength of Murashige and Skoog (MS) medium and investigating the effect of N6-benzyladenine (BA) on in vitro culture and multiplication of this plant. Clean cultures of D. garrettii with pseudobulb at the height of 1 cm were inoculated onto 3 different strengths of MS culture media (¼MS, ½MS, MS) supplemented with 1 g/L activated charcoal (AC). After 8 weeks, all explants cultured on ½MS+AC medium could survive with the highest production in number of new shoot (1.00+0.13 shoots/explant) and root (1.69+0.32 roots/explant). The pseudobulbs of regenerated shoots, having 1 cm height from ½MS+AC medium, were subsequently excised and cultured onto ½MS+AC medium supplemented with 0-8 mg/L BA. At 12 weeks, all of these explants were transferred onto BA-free ½MS+AC medium for 8 weeks. The result revealed that the ½MS+AC medium without BA was the best medium for propagating this rare orchid. Therefore, BA did not significantly affect shoot proliferation in this species. However, in vitro flowering was noticed when explants were cultured onto ½MS+AC medium supplemented with 2-4 mg/L BA. This finding may be applied for in vitro pollination which may be advantage for conservation of D. garrettii in the future. These presented consequences can be used as a basic approach for conserving this plant and may act as an additional method to sustain the biodiversity of Thailand.
2. Bai, Y., Bao, Y.H., Wang, W.Q., & Yan, Y.N. (2006). Tissue culture and rapid propagation of Dendrobium crepidatum Lindl. ex Paxt. Plant Physiology Communications, 42, R28-S68. (in Chinese).
3. Bernier, G. (2013). My favourite flowering image: the role of cytokinin as a flowering signal. Journal of Experimental Botany, 64(18), 5795-5799.
4. BGO Plant Databases, (2017). Dendrobim garrettii Seidenf. Retrieved from: http://www.qsbg.org/ database/botanic_book%20full%20option/search_detail.asp?botanic_id=3027. (in Thai).
5. Boullani, R.E., Lagram, K., Mousadik, A.E., & Serghini, M.A. (2017). Effect of explants density and size on the in vitro proliferation and growth of separated shoots of globe artichoke (Cynara cardunculus var. scolymus L.). Journal of Materials and Environmental Sciences, 8(7), 2469-2473.
6. Carimi, F., Zottini, M., Formentin, E., Terzi, M., & Lo Schiavo, F. (2003). Cytokinins: new apoptotic inducers in plants. Planta, 216(3), 413-421.
7. Chamchumroon, V. (2017). Threatened Plants in Thailand. Bangkok: Department of National Parks, Wildlife and Plant Conservation.
8. Christie, A., Jenny, K.T., & Smith, M.A.L. (1995). Automation and Environmental Control in Plant Tissue Culture. Dordrecht: Kluwer Academic Publishers.
9. Dutra, D., Johnson, T.R., Kauth, P.J., Stewart, S.L., Kane, M.E., & Richardson, L. (2008). Asymbiotic seed germination, in vitro seedling development, and greenhouse acclimatization of the threatened terrestrial orchid Bletia purpurea. Plant Cell, Tissue and Organ Culture, 94, 11-21.
10. George, E.F., Hall, M.A., & De-Klerk, G.J. (2008). Plant Propagation by Tissue Culture. (3rd ed). Dordrecht: Springer.
11. Juntab, S., Pattanakiat, S., & Sukprasert, (2013). Production orchid of farmers in Bangkok, Samut Sakhon and Nakhon Pathom Province. Journal of Agricultural Extension and Communication, 9(1), 8-16. (in Thai).
12. Kabir, M.F., Rahman, M.S., Jamal, A., Rahman, M., & Khalekuzzaman, M. (2013). Multiple shoot regeneration in Dendrobium fimbriatum Hook, an ornamental orchid. Journal of Animal and Plant Sciences, 23(4), 1140-1145.
13. Khaenthong, I. (2014). Micropropagation of varigated-leaf Dendrobium Burana Jade. Master’s Thesis, Department of Plant Science, Faculty of Science, & Department of Pharmaceutical Botany, Faculty of Pharmacy, Mahidol University.
14. Kong, Q., Yuan, S.Y., & Vegvari, G.Y. (2007). Micropropogation of an orchid Dendrobium strongylanthum Rchb.f. International Journal of Horticultural Science, 13(1), 13:61-64.
15. Kukulczanka, K., & Wojciechowska, U. (1983). Propagation of two Dendrobium species by in vitro culture. Acta Horticulturae, 131, 105-110.
16. Lee, E. J., & Paek, K.Y. (2012). Enhanced productivity of biomass and bioactive compounds through bioreactor cultures of Eleutherococcus koreanum Nakai adventitious roots affected by medium salt strength. Industrial Crops and Products, 36(1), 460-465.
17. Luo, J. P., Wang, Y., Zha, X. Q., & Huang, L. (2008). Micropropagation of Dendrobium densiflorum Lindl. ex Wall. through protocorm-like bodies: effects of plant growth regulators and lanthanoids. Plant Cell, Tissue and Organ Culture, 93, 333-340.
18. Luo, J.P., Wawrosch, C., & Kopp, B. (2009). Enhanced micropropagation of Dendrobium huoshanense C.Z. Tang et S.J. Cheng through protocorm-like bodies: the effects of cytokinins, carbohydrate sources and cold pretreatment. Scientia Horticulturae, 123, 258-262.
19. Mok, M.C., Martin, R.C., & Mok, D.W.S. (2000). Cytokinins: biosynthesis metabolism and perception. In Vitro Cellular & Developmental Biology - Plant, 36(2), 102-107.
20. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.
21. Nayak, N.R., Sahoo, S., Patnaik, S., & Rath, S.P. (2002). Establishment of thin cross section (TCS) culture method for rapid micropropagation of Cymbidium aloifolium (L.) Sw. and Dendrobium nobile Lindl. (Orchidaceae). Scientia Horticulturae, 94, 107-116.
22. Parthibhan, S., Benjamin, J.H.F., Muthukumar, M., Sherif, N.A., Kumar, T.S., & Rao, M.V. (2012). Influence of nutritional media and photoperiods on in vitro asymbiotic seed germination and seedling development of Dendrobium aqueum Lindley. African Journal of Plant Science, 6(14), 383-393.
23. Parthibhan, S., Rao, M.V., Teixeira da Silva, J.A., & Kumar, T.S. (2018). Somatic embryogenesis from stem thin cell layers of Dendrobium aqueum. Biologia Plantarum, 62, 439-450.
24. Paul, S., Kumaria, S., & Tandon, P. (2012). An effective nutrient medium for asymbiotic seed germination and large-scale in vitro regeneration of Dendrobium hookerianum, a threatened orchid of northeast India. AoB Plants, plr032, doi:10.1093/aobpla/plr032.
25. Plants of the World Online. (2020). Dendrobium Sw. Retrieved February 18, from http://plantsoftheworldonline.org/taxon/urn:lsid:ipni.org:names:325886-2.
26. Rao, S. & Barman, B. (2014). In vitro micropropagation of Dendrobium chrysanthum Wall. ex Lindl. a threatened orchid. Scholars Academic Journal of Biosciences, 2(1), 39-42.
27. Roy, J., & Banerjee, N. (2003). Induction of callus and plant regeneration from shoot-tip explants of Dendrobium fimbriatum Lindl. var. oculatum Hk. f. Scientia Horticulturae, 97, 333-340.
28. Seidenfaden, G. (1985). Orchid Genera in Thailand XII. Dendrobium Sw. Opera Botanica no. 83. Copenhagen: Council for Nordic Publications in Botany.
29. Sharma, U., Rao, V. R., Nohan, J.S.S., & Reddy, A.S. (2007). In vitro propagation of Dendrobium microbulbon A. Rich - a rare ethnomedicinal herb. Indian Journal of Biotechnology, 6, 381-384.
30. Swarts, N.D., & Dixon, K.W. (2009). Terrestrial orchid conservation in the age of extinction. Annals of Botany, 104(3), 543-556.
31. Teixeira da Silva, J.A., Cardoso, J.C., Dobranszki, J., & Zeng, S. (2015). Dendrobium micropropagation: a review. Plant Cell Reports, 34(5), 671-704.
32. Teixeira da Silva, J.A., Zeng, S., Cardoso, J.C., Dobranszki, J., & Kerbuay, G.B. (2014). In vitro flowering of Dendrobium. Plant Cell, Tissue and Organ Culture, 119(3), 447-456
33. Thomas, T. D. (2008). The role of activated charcoal in plant tissue culture. Biotechnology Advances, 26(6), 618-631.
34. Tongdonae, S., Jaichagun, M., & Sripotar, D. (2014). Study on orchid identififification of genus Dendrobium to monitor and control wild populations which impact by exportation in compliance with the Plant Act B. E. 2518. Thai Agricultural Research Journal, 32(1), 35-44. (in Thai).
35. Utami, E.S.W., Hariyanto, S., & Manuhara, Y.S.W. (2016). In vitro propagation of the endangered medicinal orchid, Dendrobium Iasianthera J. J. Sm through mature seed culture. Asian Pacific Journal of Tropical Biomedicine, 7(5), 406-410.
36. Walter, K.S., & Gillett, H.J. (1998). 1997 IUCN Red List of Threatened Plants. Compiled by the World Conservation Monitoring Centre. Norwich: IUCN - The World Conservation Union, Gland, Switzerland and Cambridge, UK.
37. Wannakrairoj, S. (2008). Status of ornamental plants in Thailand. Acta Horticulturae, 788, 29-36.
38. Winarto, B., Rachmawati, F., Santi, A., & Teixeira da Silva, J.A. (2013). Mass propagation of Dendrobium ‘Zahra FR 62’, a new hybrid used for cut flowers, using bioreactor culture. Scientia Horticulturae, 161, 170-180.
39. Winarto, B., & Teixeira da Silva, J.A. (2015). Use of coconut water and fertilizer for in vitro proliferation and plantlet production of Dendrobium ‘Gradita 31’. In Vitro Cellular & Developmental Biology - Plant, 51, 303-314.
40. Zhao, D., Hu, G., Chen, Z., Shi, Y., Zheng, L., Tang, A., & Chulin, L. (2013). Micropropagation and in vitro flowering of Dendrobium wangliangii: a critically endangered medicinal orchid. Journal of Medicinal Plants Research, 7(28), 2098-2110.
Copyright (c) 2020 Princess of Naradhiwas University Journal
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.