2.3.2 Protoplast Culture
The protoplasts were cultured in liquid medium containing varied growth regulators. Sustained divisions could
be observed in KM medium containing 1.0 mgl-1 2, 4-D, 0.1 mgl-1 NAA and 0.05 mgl-1 zeatin. All the growth
regulators used in culture were filter sterilized using sterile 0.22 μm cellulose nitrate membrane. Dilutions of the
culture media were made following the first mitotic divisions for reducing the osmotic pressure of the culture