Cost Effective In Vitro Conservation of the Threatened Malabar Daffodil Orchid using Protocorm-Like Bodies

Cost effective contamination free two-step protocol for in vitro conservation of Ipsea malabarica (Reichb. f) J.D.Hook., �The Malabar Daffodil Orchid� an endemic and endangered orchid of the Western Ghats of Kerela using protocorm-like bodies (PLBs) and the PLB-derived plantlets were achieved. Cost effectiveness and period of storage depended on level of sugar, culture vessel and closures. PLB clumps developed by transforming the axillary buds of in vitro shoots upon transfer to half-strength Murashige and Skoog (MS) medium containing 1% commercial sugar in jam bottles with autoclavable polyethylene sheets/ polypropylene caps as closures facilitated storage upto 15 months. During storage, PLBs were converted to small plantlets. Transfer of these plantlets to sugar-free(Photoautotrophic) half-strength basal medium enable storage of another 27 months. PLBs or plantlets in culture tubes with cotton plugs as closures beset with the problem of exhaustion of the medium. Nevertheless, culture tubes with polyethylene sheet closures tied with twine or rubber band did not exhibit significant difference in period of storage to that in jam bottles, but was more expensive due to the high cost of culture tubes. The cost of he culture tubes was three times higher than jam bottles. In vitro conservation of PLBs and PLB- derived plantlets in sequence facilitated a total of 42 months storage at reduced cost without the peril of contamination. Transfer of the plantlets after the storage to full strength MS medium having 13.3uM N6- MS medium containing 6.97uM kinetin the PLBs were converted to plantlets within 40 days. Repeating cycle of storage at PLBs and plantlet level, and induction of PLBs and plantlet development enables simultaneous propagation and conservation of this threatened orchid.