Cultures were established following the procedure described
earlier (Kumar et al., 1999). Cormels were descaled and washed
thoroughlyunder running tap water. These were thensurface disinfected
using mercuric chloride and inoculated on basal MS medium
(Murashige and Skoog, 1962) containing 58 mM sucrose, 0.7% (w/v)
agar, and supplemented with 5.0 M 6-benzyladenine (BA). The
pH of the medium was adjusted to 5.8 before autoclaving at 121 ◦C
for 15 min. The cultures were incubated at 25 ± 1 ◦C in 16-h light
(60 mol m−2 s−1; using white cool fluorescentlights) and 8-h dark
cycle. Actively growing shoots that sprouted from the cormels were
multiplied and maintained on MS medium supplemented with
1.0 M BA and 10.0 M -naphthalene acetic acid (NAA) and the
same were used for further experimentation.
Microshoots were sub-cultured on basal MS medium containing
232 mM sucrose. Rooting in shoots was induced by subjecting
these cultures to heat shock treatment (50 ◦C, 1-h) after one week
of subculture (Kumar et al., 1999). Well rooted shoots (plantlets)
were then used to induce cormlet formation.
2.3