2.3. Root regeneration and acclimatization
The healthy and elongated shoots were cultured on 0.8% agar-gelled
¼, ½, ¾ and full-strength MS salts supplemented with 0.2% activated
charcoal (AC) for in vitro rooting. Indole-3-butyric acid (IBA) and β-
naphthoxyacetic acid (NOA) with half and full strength of MS medium
was also evaluated for in vitro root regeneration. The rooting response
was recorded after 4 weeks. The in vitro rooted plantlets were taken
out of the culture vessels and washed thoroughly with sterile water to
remove adhered nutrient agar to avoid microbial contaminations.
These plantlets were transplanted into plastic bags containing sterile
sand and soil in the ratio of 1:1 (v/v) for subsequent hardening. Alternatively,
the healthy and elongated shoots were pulse-treated with different
concentrations of IBA (0.0–4.92 mM) and NOA (0.0–4.95 mM) for
3–5 min and transplanted into plastic bags containing sterile sand and
soil for ex vitro rooting and subsequent acclimatization. Both in vitro
rooted plantlets and pulse treated shoots for ex vitro rooting were covered
with transparent plastic bags to maintain high RH. These plantlets
were nutrified with ¼ strength of MS macro-salt solution and incubated
in a culture room.