KEYWORDS: 1-naphthaleneacetic acid (NAA) 6-Benzylaminopurine (BAP) Leaf disc Micropropagation Murashige Skoog (MS) medium

"Development of in vitro callus induction protocol for Rauvolfia serpentina",


Rauvolfia serpentina (family Apocynaceae) is a perennial undershrub and threatened plant species in Sri Lanka which is considered a therapeutically important traditional medicinal plant. Due to low seed germination and slow natural propagation, micropropagation techniques would be useful to produce healthy plant material and contribute to the ex situ conservation of R. serpentina. The objective of the study was to identify the optimisation of in vitro callus induction protocol using leaf disc explants of R. serpentina. Surface sterilization of explants were compared using varied concentrations of Clorox solution at different time intervals and 70% ethanol at fixed time period. Sterilised explants were cultured in Murashige and Skoog (MS) basal nutrient medium supplemented with different concentration combinations of 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) plant growth regulators. The cultures were incubated at 25 ± 1 °C. The survival percentage of the explants were determined after two weeks of sterilisation and the mass of callus was measured after six weeks of incubation. A 7.5% Clorox solution with a five minutes interval was found to be the optimum setting for the surface sterilisation of leaf discs and 67% of the explants survived after the sterilisation process. Rapid callus growth (2.5 ± 0.1 g) was observed in MS medium supplemented with 0.04 mg/l NAA and 2.0 mg/l BAP. Calli obtained could be used for mass propagation of R. serpentina, through indirect organogenesis.

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