Abstract

High frequency shoot regeneration from in vitro derived leaf explants of Wattakaka volubilis (L.f.) Stapf was achieved through callus mediated organogenesis. Organogenic calli were induced from 20 day old aseptic seedling explants on Murashige and Skoog medium fortified with various concentrations and combinations of plant growth regulators, benzylaminopurine (BAP), α-naphthaleneacetic acid (NAA), indole 3-butyric acid (IBA) and gibberellic acid (GA3). A mean of 8.6 shoots developed from organogenic callus induced from a 2 x 2 cm leaf explants on MS medium with 3% sucrose having 5.37µM NAA in combination with 2.22 µM BAP with 60% induction capacity. Further development of adventitious shoots could be achieved by sub culturing the callus to the same medium with 4.40 µM BAP and 0.288 µM GA3. Organogenesis could not be achieved from the calli of ex vitro derived leaf explants. The developed shoots were rooted on half-strength MS medium with 1% sucrose, 4.90 µM IBA and 0.93 µM kinetin at a frequency of 85%. Well rooted plantlets were then transplanted to vermiculite soil (3:1) mixture in polythene covered pots kept under culture room conditions. Approximately 60% plantlets survived and grew into whole plants. Key words: Adventitious shoots, caulogenesis, organogenic callus-histology.

Highlights

  • Organogenic calli were induced from 20 day old aseptic seedling explants on Murashige and Skoog medium fortified with various concentrations and combinations of plant growth regulators, benzylaminopurine (BAP), α-naphthaleneacetic acid (NAA), indole 3-butyric acid (IBA) and gibberellic acid (GA3)

  • A mean of 8.6 shoots developed from organogenic callus induced from a 2 x 2 cm leaf explants on Murashige and Skoog medium (MS) medium with 3% sucrose having 5.37μM NAA in combination with 2.22 μM BAP with 60% induction capacity

  • Wattakaka volubilis (L.f.) Stapf (Syn.: Dregea volubilis (L.f.) Benth. ex Hook.f.) (Asclepiadaceae) is an important monotypic woody twiner distributed in India, Sri Lanka, and Java

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Summary

INTRODUCTION

Wattakaka volubilis (L.f.) Stapf (Syn.: Dregea volubilis (L.f.) Benth. ex Hook.f.) (Asclepiadaceae) is an important monotypic woody twiner distributed in India, Sri Lanka, and Java. An ointment preparation with trade name “Hemajeevanthi” from leaves of this plant is commonly used for treatment of pyoderma, tineapedis, scabies, plantar psoriasis, as well as cuts and wounds (Thomas et al, 1996). Stems of this woody twiner were exploited commercially as a substitute for ropes. Other regeneration protocols reported nodal explants in W. volubilis (Yogananth et al, 2011; Vinothkumar et al, 2011). There has been no report on regeneration of W. volubilis through organogenesis and the present study describes a reproducible, efficient protocol of regeneration system by adventitious shoots using immature leaf explants. Micropropagation through adventtious shoots has been reported as an appropriate method of multiplication in many Asclepiadaceous plants (Prasad et al, 2004; Saha et al, 2003; Faisal and Anis, 2003)

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