Micropropagation of the Thai orchid <Emphasis Type="Italic">Grammatophyllum speciosum</Emphasis> blume |
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Authors: | Kathawut Sopalun Kanchit Thammasiri Keiko Ishikawa |
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Institution: | (1) Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Phyathai, Bangkok, 10400, Thailand;(2) Department of Plant Science, Faculty of Science, Mahidol University, Rama VI Road, Phyathai, Bangkok, 10400, Thailand;(3) Institute of Molecular Biosciences, Mahidol University, Putthamonthon, Nakhonpathom, 73170, Thailand;(4) Department of Research and Development, Japan Horticultural Production and Research Institute, Kamishiki, Matsudo, Chiba 270-2221, Japan; |
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Abstract: | Protocorm-like bodies (PLBs) were induced from shoot tips of Grammatophyllum speciosum, a Thai orchid. The highest frequency of PLBs (93%) were observed on explants incubated on 1/2-Murashige and Skoog (MS) liquid
medium containing 2% (w/v) sucrose without any plant growth regulators (PGRs). Tests with different carbon sources compared
to sucrose revealed that maltose promoted the highest relative growth of G. speciosum PLBs (7-fold increase), while trehalose and sucrose yielded 5-fold and 4-fold increases, respectively. In 1/2 MS liquid medium,
addition of 15 mg/l of chitosan promoted a 7-fold increase in PLB growth while 25 mg/l promoted a 4-fold increase. However,
the relative growth rate in solid culture was significantly lower than that in liquid culture. In addition, chitosan supplementation
in solid medium promoted shoot formation but not rooting. Plantlet regeneration was induced using a combination of NAA and
BA supplementation in 1/2 MS solid medium with optimum induction shoot and root formation at 2.0 mg/l NAA and 1.0 mg/l BA.
Using this protocol, approximately 8 months was required to obtain a hundred plantlets from one shoot tip. The plantlets showed
no changes in ploidy when tested by flow cytometry. |
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