Abstract:
Agarwood produced by Gyrinops walla Gaertner, an
endemic plant species of Sri Lanka, arguably the costliest
wood, is used as a major constituent in cosmetic industries.
Application of plant cell culture technique serves as the
best alternative for the production of fragrance
compounds of G. walla while ensuring the sustainability of
the species. Using the advantages of plant cell culture and
Thin Layer Chromatography (TLC), a preliminary study
was carried out to compare the phytochemical
constituents of callus and plant extracts (i.e stem, leaves,
bark and agarwood) of G. walla. Different callus lines were
developed on MS and modified MS media with reduced
level of NH4NO3, supplemented with different
combinations and concentrations of plant growth
regulators and multiplied. Freeze dried samples were
extracted with hexane, ethyl acetate and methanol and run
on a TLC plate under different eluting solvent systems. A
higher number of TLC spots were observed in the ethyl
acetate extracts of all samples, under 15 percent methanol:
chloroform solvent system. Based on TLC chemical profiles
of all callus lines, MS media added with 1.0 mg/L BAP, 0.5
mg/L kinetin and 0.1 mg/L 2,4-D and ½ MS contained 1.0
mg/L BAP and 0.1 mg/L 2,4-D were selected as best lines.
The extracts of the selected two callus lines matched with
the fingerprint profiles of best plant samples i.e stem, bark
and agarwood. Compounds present in the extracts of callus
samples, plant samples and agarwood, appearing at
Rf=0.56, 0.74 and 0.78, could be used as possible markers
for G. walla. The chromatography profiling by TLC has
proven the presence of similar phytochemical constituents
in callus, plant samples and agarwood. Therefore, it can be
concluded that plant cell culture technique guarantees the
possibility to produce same compounds that are present in
plant and agarwood of G. walla.
