Int. J. Biosci.2( 3), 97-105, March 2012
Cell suspension culture from leaf-derived callus of Bitter melon (Momordica charantia L.) was established. The callus could be induced from leaf segments on agarified Murashige and Skoog (MS) medium containing 1.0 mgl-1 2,4-dichlorophenoxyacetic acid (2,4-D), which consequently used for suspension culture. In the establishment of cell suspension culture, the highest performance in the growth of cells was observed in liquid MMS (addition of 0.5 mgl-1 folic acid and 0.05 mgl-1 biotin to MS medium) medium containing 2,4-D (1.5 mgl-1) when callus was subcultured. The S-shaped growth curve with three typical phages (lag, exponential and stationary phases) was found in the batch culture. Cells in suspension culture underwent division as result both free cells and cell aggregates were formed. The number of cell aggregation affected by the initial amount of cells. When the initial amounts of cell were increased from 1 to 5 ml sedimented cell volume (SCV), the number of cell aggregates was increased gradually. The number of aggregates decreased with further increasing initial amount of cells from 5 ml SCV. Aggregate formation at a highest level when 5 ml (SCV) initial cells subcultured. The duration of subculture was found to be effective for cell proliferation in suspension and proliferated cell structure. The cell growth in culture was lower when suspension maintained up to 8 weeks without subculturing. The elliptical cell structure was mostly found in suspension maintained up to 8 weeks without subculturing. The most of cells were spherical-shaped and cell growth was high in suspension when suspension culture changed to fresh medium at 7-week-interval. The established system on the cell suspension culture might be used for establishing an efficient somatic embryogenesis method of bitter melon.
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