2.2 Regeneration Experiments
5-d old seedlings were used as explants in regeneration experiments. The cotyledons and the apical bud were
excised and separated from the hypocotyl region and hypocotyl explants (1.0 cm long) were cultured on MS
medium containing different combinations and concentrations of plant growth regulators (PGRs).
2.2.1 Culture Media and Culture Condition
Required PGRs were added to MS basal medium (Murashige & Skoog, 1962) and the pH was adjusted to 5.8
before autoclaving at 121°C for 20 min. Explants were grown on medium solidified with 8.0 gl-1 agar (Himedia,
Mumbai, India) and kept under 16 h photoperiod, illuminated with cool white fluorescent lamps at 28°C. MS
basal medium was supplemented with 0.5 - 7.5 mgl-1 6-BAP (6-Benzylaminopurine) in combination with 25 or
50 mgl-1 adenine hemi sulfate (AdSO4) for callus induction and subsequent regeneration. The response was
carefully monitored and scored.
Calli bearing shoot clumps were sub-cultured to medium containing 0.1 mgl-1 NAA and 1.0 mgl-1 6-BAP for
rapid shoot elongation. The number of elongated shoots was visually examined and the shoots were separated
from the clump.