Vanda tessellata (Roxb.) Hook. ex G. Don. (grey orchid, family Orchidaceae) is an epiphytic orchid of horticultural importance and currently under threat due to overharvesting and habitat destruction. Micropropagation protocols were developed for the production of grey orchid but the survival success of in vitro regenerated plantlets is uncertain due to lack of understanding about the adaptation mechanism during hardening. The present study describes the structural adaptation mechanism of V. tessellata when the in vitro regenerated plantlets were acclimatized under the greenhouse conditions. Light microscopy has been implicated to identify the adaptational alterations during in vitro to ex vitro transition of micropropagated plantlets.
Oldenlandia umbellata L. gains importance due to its medicinal properties and the presence of anthraquinones based natural dyes in the roots. Present study describes the effect of Murashige and Skoog's (MS) liquid medium (full strength) on in vitro regeneration, flower bud induction and ex vitro rooting in O. umbellata. Shoot segments with 2-3 nodes (each node with 2 axillary buds) served as explants for establishment of cultures. The liquid medium augmented with 2.0 mg L -1 6-benzylaminopurine (BAP) with additives (50 mg L -1 of ascorbic acid and 25 mg L -1 each of arginine, adenine sulphate and citric acid) was effective for shoot bud induction (6.4±0.19 shoots per explant within 2-3 weeks). The shoots were further multiplied (89.3±1.07 shoots, 2-3 weeks) when the shoot clusters obtained from the culture initiation directly transferred to the full-strength MS liquid medium incorporated with 1.0 mg L -1 BAP and 0.5 mg L -1 indole-3 acetic acid (IAA) with additives. Flower buds were induced (12.0±0.15 buds per shoot) when the shoots were cultured on 1.0 mg L -1 BAP and kinetin (Kin, 6-furfurylaminopurine) and 0.5 mg L -1 of IAA at 45 µmol m −2 s −1 SFPD (Spectral Flux Photon Density) light intensity for 14/10h (light/dark) photoperiod. The adventitious roots were induced on 1/4 strength MS medium supplemented with 1.5 mg L -1 indole-3 butyric acid (IBA). Ex vitro rooting was achieved (16.0±0.53 roots per shoot) by pulse treatment of the shoots with 300 mg L -1 IBA for 2 min. The in vitro produced plantlets were acclimatized in the greenhouse and finally translocated to the in vivo conditions with 93 % success rate. This is the foremost (use of liquid MS medium) and cost-effective method for large scale multiplication of O. umbellata.
Oldenlandia umbellata L. gains importance due to its medicinal properties and the presence of anthraquinones based natural dyes in the roots. Present study describes the effect of Murashige and Skoog’s (MS) liquid medium (full strength) on in vitro regeneration, flower bud induction and ex vitro rooting in O. umbellata. Shoot segments with 2-3 nodes (each node with 2 axillary buds) served as explants for establishment of cultures. The liquid medium augmented with 2.0 mg L-1 6-benzylaminopurine (BAP) with additives (50 mg L-1 of ascorbic acid and 25 mg L-1 each of arginine, adenine sulphate and citric acid) was effective for shoot bud induction (6.4±0.19 shoots per explant within 2-3 weeks). The shoots were further multiplied (89.3±1.07 shoots, 2-3 weeks) when the shoot clusters obtained from the culture initiation directly transferred to the full-strength MS liquid medium incorporated with 1.0 mg L-1 BAP and 0.5 mg L-1 indole-3 acetic acid (IAA) with additives. Flower buds were induced (12.0±0.15 buds per shoot) when the shoots were cultured on 1.0 mg L-1 BAP and kinetin (Kin, 6-furfurylaminopurine) and 0.5 mg L-1 of IAA at 45 µmol m−2 s−1 SFPD (Spectral Flux Photon Density) light intensity for 14/10h (light/dark) photoperiod. The adventitious roots were induced on 1/4 strength MS medium supplemented with 1.5 mg L-1 indole-3 butyric acid (IBA). Ex vitro rooting was achieved (16.0±0.53 roots per shoot) by pulse treatment of the shoots with 300 mg L-1 IBA for 2 min. The in vitro produced plantlets were acclimatized in the greenhouse and finally translocated to the in vivo conditions with 93 % success rate. This is the foremost (use of liquid MS medium) and cost-effective method for large scale multiplication of O. umbellata.
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