Biosynthetic potential of in vitro grown callus cells of Cassia senna L. var. senna.

摘要

A protocol for tissue culture of Cassia senna is established in different morphogenetic media and in vitro-grown tissues/cells were analysed for their biosynthetic potential. Authenticated seeds from Gujarat, India, were washed, sterilized and inoculated for germination in axenic conditions. Cotyledonary leaves of aseptically grown seedlings were cut and inoculated onto MS medium supplemented with different concentrations and combinations of growth regulators (e.g. benzyladenine, adenine sulfate and coconut milk (CM)) for shoot differentiation induction. The compact non-morphogenic callus mass proliferated during shoot regeneration phase was analysed for total sennoside content (hydroxyanthracene derivatives (HAD) calculated as sennoside B). Analysis of the above cell masses showed sennoside presence in all samples tested, indicating that these in vitro-cultured cells of callus inherited the same genetic makeup as the cells of mother plant, and hence have the potential to synthesize secondary metabolites. Sennoside content was maximum (0.13%) in the medium supplemented with 16 micro M benzyladenine and 10% CM, and minimum (0.06%) in the medium supplemented with 14 micro M benzyladenine and 8 micro M adenine sulfate. In other media, sennoside content varied from 0.08 to 0.10%. The medium supporting minimum organogenesis showed minimum biosynthesis of sennosides. In the medium with 16 micro M benzyladenine+10% CM, along with shoot-forming nodules, non-morphogenic compact cell mass was also present, which failed to produce shoot primordia. The results indicated that the in vitro-cultured partially organized cells of C. senna inherited the biosynthetic potential which can be exploited for the large-scale production of sennosides under proper growth conditions..