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Optimization of Embryogenic Callus Induction and Plant Regeneration in Orchid Coelogyne cristata
Aung Htay Naing,Ki Byung Lim 한국원예학회 2011 원예과학기술지 Vol.29 No.3
An efficient protocol was established for high frequency somatic embryogenesis through a callus culture of Coelogyne cristata. The best frequency of callusing was obtained from a PLB segment (3-5 ㎜) cultured on MS medium supplemented with coconut water (CW) and a combination of both 3 mg · ℓ?¹ of 2,4-D and BA. When the calli were sub-cultured on the MS medium without any PGRs, the average number of somatic embryos were higher than those with PGRs treatment. NAA is the most critical factor among PGRs, which dramatically hindered for the formation of a somatic embryo. The efficacy of the addition of coconut powder (CP) for somatic embryogenesis was almost the same in all treatments. However, the number of somatic embryos formed distinctly depended on age of the callus. The somatic embryos converted into healthy plants with well-developed shoots on the same medium. Plantlets showed the best responses of root and shoot growth when transferred to ½ MS medium containing 1.5 g · ℓ?¹ of activated charcoal. All plants with above 3.0-㎝-high were successfully acclimatized in the greenhouse.
Micropropagation and Conservation of the Wild Medicinal Orchid, Coelogyne cristata
Aung Htay Naing,Khin Thida Myint,Yoon Jung Hwang,In Sook Park,Jae Dong Chung,Ki Byung Lim 한국원예학회 2010 Horticulture, Environment, and Biotechnology Vol.51 No.2
Micropropagation of the wild medicinal orchid, Coelogyne cristata, was conducted through culture of shoot axis, intact seedlings and pseudobulb segments. The shoot axis cultured on half strength Murashige and Skoog (½ MS) medium with 0.44 ㎎?L<SUP>-1</SUP> thidiazuron (TDZ) gave the optimum shoot response and fresh weight. Higher concentrations of TDZ above the optimum level seemed to gradually increase inhibitory effects on shoot regeneration and plant growth. Intact seedlings showed maximum number of shoots on Murashige and Skoog (MS) medium with 1.0 ㎎?L<SUP>-1</SUP> naphthalene acetic acid (NAA) and 1.0 ㎎?L<SUP>-1</SUP> 6-benzylaminopurine (BA) combination, without activated charcoal (AC). In two types of pseudobulb segment (basal and terminal segments) culture, basal segments produced better shoot response in all treatments than that of terminal segments. Of different inoculum types, medium and growth regulators used in the present study, the culture of seedlings on MS media with 1.0 ㎎?L<SUP>-1</SUP> NAA and 1.0 ㎎?L<SUP>-1</SUP> BA combination without AC appears to have great potential for in vitro propagation of this extremely wild orchid. The obtained plantlets showed no genetic variation.