In vitro regeneration frequency, micro-morphological studies and ex vitro rooting of Hemidesmus indicus (L.) R. Br.: a multi-potent endangered climber
Tóm tắt
Hemidesmus indicus is an endangered medicinal plant of Peninsular India. A rapid micropropagation method using nodal segments of a 4 year old plant has been achieved in the present study. The nodal explants (2.0–3.0 cm long) were harvested from actively growing shoots of conventionally raised plants and cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg l−1 6-benzylaminopurine (BAP). Hundred percent explants responded with 5.2 ± 0.25 cm long multiple shoots (9.0 ± 0.53) from the nodal meristems. The shoots were further multiplied enormously (272 ± 4.12 shoots per explant) by repeated subculture of mother explants and freshly induced shoot clumps on MS medium with the reduced concentrations of BAP (1.0 mg l−1), kinetin (Kin; 0.5 mg l−1) and indole-3 acetic acid (IAA; 0.1 mg l−1) within 4 weeks. Highest root induction (62.0 ± 0.54 roots per shoot) was observed on 1/4th strength MS medium supplemented with 3.0 mg l−1 indole-3 butyric acid (IBA). About 96 % shoots were rooted (45.0 ± 0.48 roots per shoot) by ex vitro methods when the cut ends of the shoots pulse treated with IBA (400 mg l−1) for 5 min. The in vitro as well as ex vitro rooted plantlets were acclimatized successfully in the greenhouse. Ex vitro rooted plantlets exhibited higher percentage (98 %) of survival as compared to the in vitro rooted plantlets (91 %) in the field conditions. There were not any observable differences between in vitro propagated and field transplanted plantlets. A comparative foliar micro-morphological study of H. indicus was conducted to understand the micro-morphological changes in the plants while shifting from in vitro to the in vivo conditions in terms of variations in stomata, venation pattern and vein density, and trichomes. The aforementioned protocol could be successfully used for the large-scale multiplication and conservation of germplasm of this endangered plant species.
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