Efficient in vitro Regeneration Protocol as an Aid to Conserve Wild Population of Centella asiatica (L.) Dunal
B.G. Hanumantharaya
Plant Tissue Culture Laboratory, Department of Horticulture, University of Agricultural Sciences GKVK Campus, Bengaluru- 560 065, Karnataka, India
B.N. Sathyanarayana
Plant Tissue Culture Laboratory, Department of Horticulture, University of Agricultural Sciences GKVK Campus, Bengaluru- 560 065, Karnataka, India
Waman Ajit Arun
Plant Tissue Culture Laboratory, Department of Horticulture, University of Agricultural Sciences GKVK Campus, Bengaluru- 560 065, Karnataka, India
*Author to whom correspondence should be addressed.
Abstract
In order to reduce the pressure on the wild stock of Centella, which is being exploited to meet the ever increasing demand from pharmaceutical industries, an efficient in vitro multiplication protocol has been developed. The aseptic cultures were initiated using nodal explants on Murashige and Skoog�s media and were subjected to various kinds and concentrations of the growth regulators for multiplication. Benzylamino purine (BAP) was found to be better source of cytokinin than Kinetin (Kn), when used alone or in combination with auxins. Explants were best initiated on media supplemented with 2 mg/l BAP + 0.1 mg/l NAA, wherein maximum shoot induction percentage (96.67%), quicker induction response (5.333 days), maximum multiple shoots production (10.333) and highest shoot length (3.333 cm) were noticed. Induced shoots showed maximum proliferation (40 shoots/explant) on the media supplemented with 2 mg/l BAP. All rooting parameters were found to be superior on the quarter strength MS medium than on half strength medium. A combination of 0.3 mg/l NAA + 0.3 mg/l IBA was optimum for getting the best response towards rooting. Plantlets were acclimatized with 97 percent success when vermicompost and sand (4:1) was used as a hardening medium.
Keywords: Direct regeneration, medicinal plant, threatened species, node, plant growth regulators