Rooting and acclimatization of regeneration plants
Regenerated shoots (around 1cm long) were placed in MS
medium. The medium was solidified with 3g/l Phytagel and
dispensed at 30ml per Magenta box and four shoots were
cultured in each box. Regenerated shoots were incubated at
25±1 °C in a growth chamber with a 16-h photoperiod under
standard cool white fluorescent tubes (35 μmol s-1 m-2) for 5
weeks. After five weeks, the rooted plants were washed with
sterile water to remove Phytagel, transferred to pots containing
autoclaved vermiculite, and covered with polyethylene bags for
one week to maintain high humidity. The plants were then
transferred to soil and maintained in a growth chamber with a
16-h photoperiod, and a night/day temperature of 18/20°C for 2
weeks. These hardened plants then transferred to the
greenhouse.