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VOL. 11, ISSUE 3 (2026)
Establishment of an efficient in vitro seed germination and micropropagation protocol for Dendrobium aggregatum Roxb.: A medicinally important orchid of Bangladesh
Authors
Md Mustakim Zulfikar Alam, Md Mahbubur Rahman, Tapash Kumar Bhowmik
Abstract
Dendrobium aggregatum
Roxb. is an epiphytic medicinal and ornamental orchid indigenous to the
Chittagong Hill Tracts of Bangladesh. Due to habitat destruction, over
exploitation and the species' intrinsic biological limitations specifically the
lack of endosperm and absolute dependency on mycorrhizal fungi for natural seed
germination wild populations are rapidly declining. This study was undertaken
to establish a reproducible, highly efficient in vitro seed germination
and direct micropropagation protocol for the conservation and mass
multiplication of D. aggregatum. Immature capsules were aseptically
cultured on four basal media formulations: Knudson C (KC), Murashige and Skoog
(MS), Phytamax (PM) and Modified Vacin and Went (MVW), evaluated with and
without plant growth regulators (PGRs). Full strength MS medium supplemented
with 0.5 mg/l 6-Benzylaminopurine (BAP) and 0.5 mg/l α-Naphthaleneacetic acid
(NAA) exhibited absolute superiority, yielding a 100.00% germination rate and
initiating early protocorm development within 10.13 ± 0.41 weeks. Subsequent
complete plantlet development occurred in 25.53 ± 0.56 weeks. For rapid shoot
elongation, transferring germinated seedlings to MS medium fortified with 1.5
mg/l Kinetin (Kn) and 0.9 mg/l Indole-3-acetic acid (IAA) induced a maximum
longitudinal increase of 2.88 ± 0.04 cm over 30 days. High frequency
micropropagation via direct organogenesis was optimally achieved using
nodal segments on MS medium enriched with 2.0 mg/l BAP and 1.0 mg/l NAA,
delivering 72.47 ± 1.56% response and 4.8 ± 0.42 multiple shoot buds (MSBs) per
explant in 8.77 ± 0.18 weeks. Conversely, indirect morphogenic responses via
protocorm-like bodies (PLBs) were optimally induced from leaf segments using
2.0 mg/l BAP and 1.0 mg/l IAA (50.00 ± 1.43%). Vigorous rhizogenesis was
obtained on MS medium utilizing a synergistic combination of 1.0 mg/l Indole-3-butyric
acid (IBA) and 1.0 mg/l NAA, yielding an average of 5.25 ± 0.37 roots per shoot
with a mean length increase of 3.43 ± 0.05 cm. Following sequential ex vitro
acclimatization in a substrate mix of coir, sawdust, coal, small bricks and
activated charcoal, the seedlings demonstrated a robust 80% survival rate. This
optimized protocol provides a reliable framework for the largescale commercial
cultivation and ex situ conservation of D. aggregatum.
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Pages:30-37
How to cite this article:
Md Mustakim Zulfikar Alam, Md Mahbubur Rahman, Tapash Kumar Bhowmik "Establishment of an efficient <i>in vitro</i> seed germination and micropropagation protocol for <i>Dendrobium aggregatum</i> Roxb.: A medicinally important orchid of Bangladesh". International Journal of Advanced Research and Development, Vol 11, Issue 3, 2026, Pages 30-37
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