Tissue Culture Essay

Kaempferia galanga Linn. (Family: Zingiberaceae) is distributed mainly in South East Asia and China (Sala, 1995). K. galanga is considered as a main ingredient of many traditional Ayurvedic drug preparations in Asian countries (Ibemhal et al; 2012). Essential oil extracted from rhizome is volatile (Wong et al., 1992) and is used as a spice and also used in beverage, perfume and cosmetic industries for its aroma and flavor (Arambewela and Silva, 1999). Its leaves and flowers contain flavonoids (Ghani, 1998). Main constituents of the rhizome extract contain ethyl-p-methoxycinnamate, ethylcinnamate, 3-carene, camphene, borneol, cineol, kaempferol and kaempferide which are responsible for many properties of K. galanga (Ibemhal et al; 2012). The

Thus, comparison of phytochemicals present in rhizomes of natural plants and tissue cultured plants are necessary if the tissue cultured plants are using as alternative to natural plants.
Materials and Methods
For all tissue culture experiments, Murashige and Skoog (MS) medium (Murashige and Skoog, 1962) was used as the basal medium with 30.0 g/L sucrose and jelly moss (8.0 g/L) as gelling agent. The pH of the media was adjusted to 5.8. Unless otherwise stated there were twenty replicates in each treatment. Randomized Block Design (RDB) was used in all experiments and culture bottles were randomized at seven day intervals. Cultures were incubated at 25±1ºC in 16 hours photoperiod. Results were analyzed using Minitab statistical package.
In vitro shoot induction and multiplication of K. galanga from axillary buds
Rhizomes of K. galanga was wrapped in wet tissues and allowed the axillary buds to be elongated. Axillary buds were removed carefully from the rhizome and initially washed in 5% Clorox (5.25% NaOCl™) for 15 minutes and transferred into laminar flow cabinet. Then they wrer washed in 10% Clorox and 70% ethanol each followed by two successive washings in sterile distilled water before being transferred in to culture