Abstract
This investigation was undertaken to establish an efficient protocol for virus-free plant regeneration in Coccinia grandis L. through shoot apical meristem culture. Murashige and Skoog (MS) basal medium supplemented with different concentrations and combinations of 6-benzyl amino purine (BAP), gibberellic acid (GA3), naphthalene acetic acid (NAA), and indole-3-butyric acid (IBA) was used for meristem establishment, shoot regeneration, and root induction as well as elongation. MS liquid medium supplemented with 1.0 mg l-1 BAP + 0.10 mg l-1 NAA was found to be the best medium for the primary establishment of meristems. MS medium containing 1.5 mg l-1 BAP + 0.5 mg l-1 GA3 + 0.5 mg l-1 NAA was found to be best for shoot regeneration percentage at 100.0 ± 0.0 and multiplication with 10.0 ± 0.8 shoots per meristem as well as shoot elongation (highest 9.0 ± 0.0 cm). In vitro grown shoots were subcultured and rooted with 11.0 ± 0.8 roots per shoot subsequently on MS medium containing 0.5 mg l-1 IBA. Well-rooted plantlets were gradually acclimatized and successfully established in the field condition with 100% survival rate.
Highlights
Coccinia grandis L. is an important medicinal and vegetable crop plant belonging to the family of Cucurbitaceae
Primary Establishment of Meristems Isolated meristems (Fig. 1A) were cultured (Fig. 1B) on paper-bridge used liquid Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of benzyl amino purine (BAP), naphthalene acetic acid (NAA) and GA3
The highest percentage of meristem culture response was 96.6 6 0.8 in MS liquid medium containing 1.0 mg l21 BAP 1 0.10 mg l21 NAA followed by 90.0 6 0.8% in the semisolid medium contained the same hormonal supplements
Summary
Coccinia grandis L. is an important medicinal and vegetable crop plant belonging to the family of Cucurbitaceae. At the advent of plant biotechnology, meristem culture offers a novel tool for the production of virus-free plants. There are many reports on meristem culture obtaining virus-free plants, including Helicteres isora [7], snake gourd [8], purple passion fruit [9], pumpkin [10], orchid [11], cowpea [12], and bitter gourd [13]. Rapid in vitro multiplication for virus-free plantlets of C. grandis is required to overcome its extinction. We established an in vitro virus-free plantlets propagation protocol for C. grandis and their successful acclimatization in the field condition
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