Plantlet Regeneration and Multiple Shoot Induction from Protocorm-Like Bodies (PLBs) of Medicinal Orchid Species, Dendrobium crumenatum Sw.
Keywords:Dendrobium crumenatum, Germplasm conservation, In vitro conservation, Micropropagation, Thidiazuron
To investigate the suitable medium for in vitro shoot regeneration and plantlet growth of Dendrobium crumenatum Sw., individual protocorm-like bodies (PLBs) (about 4 - 5 mm in diameter) of Dendrobium crumenatum Sw. derived from MS medium supplemented with 0.5 mg/L TDZ for 60 days of culture were cultured on 6 culture media; Murashige and Skoog (MS) medium, MS medium supplemented with 15 % (v/v) CW, MS medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC, Vacin and Went (VW) medium, VW medium supplemented with 15 % (v/v) CW, VW medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC. After 4 months of culture, MS medium containing 15 % coconut water (CW) gave the highest percentage of shooting and number of shoots per explant of 96.0 and 9.5, respectively with a significant difference from other media. The addition of 0.2 % (w/v) activated charcoal (AC) significantly increased the number of leaves and roots. PLBs developed into complete plantlets. MS medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC and VW medium supplemented with 15 % (v/v) CW and 0.2 % (w/v) AC gave the highest number of roots per plantlet and root length at 5.3 roots and 34.9 mm, respectively. After the transfer of rooted shoots to the greenhouse, 95 % of the regenerated plantlets survived and grew vigorously. Plantlets grown in vitro were successfully acclimatized in the greenhouse and showed normal development.
A Cronquist. An integrated system of classification of flowering plants. Columbia University Press, Ithaca, 1981, p. 248-50.
RL Dressler. The orchids: Natural history and classification. Harvard University Press, Cambridge, 1981, p. 332.
CJ Goh, MS Strauss and J Arditti. Floral induction and physiology in orchids. In: J Arditti (Ed.). Orchid biology: Reviews and prospectives. Cornell University Press, Ithaca, 1982, p. 213-41.
RE Holttum. The story of the pigeon orchid. In: RE Holttum (Ed.). Plant life in Malaya. Longmans, UK, 1964, p 121-41.
UM Sandrasagaran, S Subramaniam and V Murugaiyah. New perspective of Dendrobium crumenatum orchid for antimicrobial activity against selected pathogenic bacteria. Pak. J. Bot. 2014; 46, 719-24.
U Meesawat and K Kanchanapoom. In vitro plant regeneration through embryogenesis and organogenesis from callus culture of pigeon orchid (Dendrobium crumenatum Sw.). Sci. Tech. Asia 2002; 7, 9-17.
S Rittirat, K Thammasiri, S Kalawong and S Klaocheed. Ex-situ conservation of endangered medicinal Orchid species, Dendrobium crumenatum Sw. by micropropagation. In: Proceedings of the 13th International Conference ASIAN Community Knowledge Networks for the Economy, Society, Culture, and Environmental Stability. Miyazaki, Japan, 2018, p. 46-52.
T Murashige and F Skoog. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 1962; 15, 473-97.
EF Vacin and FW Went. Some pH changes in nutrient solutions. Bot. Gaz. 1949; 110, 605-17.
JM Bonga and PV Aderkas. In vitro culture of trees. Kluwer Academic Publishers, Dordrecht, 1992, p. 236.
J Arditti and R Ernst. Micropropagation of orchids. Vol II. Wiley, New York, 1993, p. 1560.
S Ichihashi and MO Islam. Effects of complex organic additives on callus growth in three orchid genera, Phalaenopsis, Doritaenopsis and Neofinetia. J. Jpn. Soc. Hortic. Sci. 1999; 68, 269-74.
RLM Pierik, PA Sprenkels, B van der Harst and QG van der Meys. Seed germination and further development of plantlets of Paphiopedilum ciliolare Pfitz in vitro. Sci. Hortic. 1988; 34, 139-53.
S Zeng, K Wu, JA Teixeira da Silva, J Zhang, Z Chen, N Xia and J Duan. Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh., an endangered terrestrial orchid. Sci. Hortic. 2012; 138, 198-209.
Y Zhang, YI Lee, L Deng and S Zhao. Asymbiotic germination of immature seeds and the seedling development of Cypripedium macranthos Sw., an endangered lady’s slipper orchid. Sci. Hortic. 2013; 164, 130-6.
CC Chu and KW Mudge. Effects of prechilling and liquid suspension culture on seed germination of the yellow lady’s slipper orchid (Cypripedium calceolus var. pubescens). Lindleyana 1994; 9, 153-9.
YI Lee, N Lee, EC Yeung and MC Chung. Embryo development of Cypripedium formosanum in relation to seed germination in vitro. J. Am. Soc. Hortic. Sci. 2005; 130, 747-53.
KP Martin and J Madassery. Rapid in vitro propagation of Dendrobium hybrids through direct shoot formation from foliar explants, and protocorm-like bodies. Sci. Hortic. 2006; 108, 95-9.
B Pant and R Gurung. In vitro seed germination and seedling development in Aerides odorata Lour. J. Orchid Soc. India. 2005; 19, 51-5.
SK Bhadra and MM Hossain. Introduction of embryogenesis and direct organogenesis in Micropera pallida Lindl. An epiphytic orchid of Bangladesh. J. Orchid Soc. India. 2004; 18, 5-9.
S Pradhan. 2007, Ex situ conservation of two orchid species viz. Cymbidium elegans Lindl. and Dendrobium densiflorium Lindl. by tissue culture technique. Ph.D. Dissertation, Central Dept Bot TU Kathmandu, Nepal.
S Asghar, T Ahmad, IA Hafiz and M Yaseen. In vitro propagation of orchid (Dendrobium nobile) var. Emma White. Afr. J. Biotechnol. 2011; 10, 3097-103.
NV Ket, EJ Hahn, SY Park, D Chakrabarty and KY Peak. Micropropagation of an endangered orchid Anoectochilus formosanus. Biol. Plant. 2004; 48, 339-44.
E Dumas and O Monteuuis. In vitro rooting of micropropagated shoots from juvenile and mature Pinus pinaster explants-influence of activated charcoal. Plant Cell Tiss. Org. Cult. 1995; 40, 231-5.
G Fridborg, ML Pedersen and T Eriksson. The effect of activated charcoal on tissue cultures: Adsorption of metabolites inhibiting morphogenesis. Physiol. Plant. 1987; 43, 104-6.
G Fridborg and T Eriksson. Effects of activated charcoal on morphogenesis in plant tissue cultures. Physiol. Plant. 1975; 34, 306-8.
MJ Constantin, RR Henke and MA Mansur. Effect of activated charcoal on callus growth and shoot organogenesis in tobacco. Vitro Cell Dev. Biol. Plant. 1977; 13, 287-93.
SJ Nissen and EG Sutter. Stability of IAA and IBA in nutrient medium to several tissue culture procedures. Hortic. Sci. 1990; 25, 800-2.
MA Weatherhead, J Burdon and GG Henshaw. Some effects of activated charcoal as an additive to plant tissue culture media. Z Pflanzenphysiol. 1978; 89, 141-7.
R Ernst. The use of activated charcoal in asymbiotic seedling culture of Paphiopedilum. Am. Orchid Soc. Bull. 1974; 43, 35-8.
R Ernst. Studies in asymbiotic culture of orchids. Am. Orchid Soc. Bull. 1975; 44, 12-8.
L Johansson, E Galleberg and A Gedin. Correlation between activated charcoal, Fe EDTA and other organic media ingredients in cultures of anthers of Anemone canadensis. Physiol. Plant. 1990; 80, 243-9.
J Van Huylenbroeck, A Piqueras and P Debergh. Photosynthesis and carbon metabolism in leaves formed prior and during ex vitro acclimatization of micropropagated plants. Plant Sci. 1998; 134, 21-30.
LP Diaz, JJ Namur, SA Bollati and OEA Arce. Acclimatization of Phalaenopsis and Cattleya obtained by micropropagation. Rev. Colomb. Biotecnol. 2010; 12, 27-40.
EF George and PC Debergh. Micropropagation: Uses and methods. In: EF George, MA Hall and GJ de Klerk (Eds.). Plant propagation by tissue culture. 3rd eds. Springer, Dordrecht, Netherlands, 2008, p. 29-64.
S Chugh, S Guha and IU Rao. Micropropagation of orchids: A review on the potential of different explants. Sci. Hort. 2009; 22, 507-20.
How to Cite
Copyright (c) 2020 Walailak University
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.