Micropropagation and Assessment of Genetic Homogeneity in Dendrobium moniliforme (L.) Sw. using Molecular Markers
Ghimire, M.; Thapa, M.; Joshi, P. R.; Paudel, M. R.; Pant, K. K.; Gurung, P.; Wagner, S. H.; Pant, B.
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Dendrobium moniliforme (L.) Sw., is a valuable medicinal orchid widely used as tonic. This study aimed to develop an effective protocol for micropropagation of D. moniliforme and to compare the genetic similarities of the micro-propagated plants to the wild plants using Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) molecular markers. Healthy seeds were cultured on Murashige-Skoog (MS) medium, supplemented with various concentrations of auxins, cytokinins and coconut water for shoot and root development. Genetic homogeneity was assessed using ten primers of each RAPD and ISSR marker. MS media fortified with 10% coconut water was efficient for seed germination and protocorm development. Maximum number of shoots were produced on half strength MS (HMS) medium supplemented with 0.25 mg/L NAA and 10% coconut water (CW) i.e 22 shoots. FMS supplemented with 0.5 mg/L indole butyric acid (IBA) was found to be best as plant hormone for root induction, as it induced ten roots/shoot. RAPD and ISSR amplified a total of 15 and 5 homogeneous DNA fragments. This protocol demonstrates a reliable and efficient method for large-scale propagation and conservation of D. moniliforme, supporting its commercial and medicinal value. HighlightsDendrobium moniliforme was micro-propagated from seeds using MS medium. RAPD and ISSR markers confirmed its genetic stability. The study supports large-scale true-to-type propagation and conservation of these valuable medicinal orchids.
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