Keywords:
In vitro, Nodal buds, 6-Benzylaminopurine, Naphthalene acetic acid, Clonal fidelity assessment
The present study reports development of an improved in vitro regeneration protocol of Strychnos potatorum L. f. a wild threatened medicinal tree species. The higher rate of morphogenetic plant regeneration was observed in seedlings derived young nodal bud explants when compared to shoot tips explants. Nodal explants cultured on MS medium supplemented with 1.0 mg/L–1 6-benzylaminopurine (BAP) in combination with 0.5 mg/L–1 naphthalene acetic acid (NAA) induced a maximum number of shoots of (10.5 ± 0.53) with a mean shoot height of (5.2 ± 0.36 cm) was obtained. BAP was found more significant in multiple shoot induction when compared to other cytokinins used in this study. In vitro rooting was achieved on half-strength MS medium augmented with different concentrations of auxins. NAA at 0.5 mg/L–1 produced the highest number of healthy roots (8.6 ± 0.70). The rooted plantlets were successfully hardened in the paper cups containing farmyard manure and garden soil in 1:1 ratio and successfully transferred to field conditions. About 80% of the plants survived in the natural habitat. The field-grown plants showed similarity in its growth characteristics to their mother plant. An assessment of clonal fidelity of regenerated plants was undertaken using inter simple sequence repeat (ISSR) markers which revealed that they are monomorphic and more comparable to their mother plants. This efficient reproducible protocol can be useful for the production of true-to-type nature and ex situ conservation of this threatened S. potatorum tree species.
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The authors are grateful to Prof. S. Karuppusamy, Department of Botany, The Madura College (Autonomous), Madurai, Tamil Nadu, India for helping in the plant collection and identification of the plant. Dr. P. Kannan thanks University Grants Commission [MRP (42/936) 2013-2017] New Delhi for financial assistance.