2.3. Callus induction
Cotyledon and hypocotyl explants were used for callus
induction. Explants were excised from seven days old in vitro
raised seedlings. Cotyledon explants were cut into small squares
and hypocotyls were cut into 5 mm length and kept on MS salts
with B5 vitamins medium fortified with various concentrations
of auxins and cytokinins. The explants were placed in a
15
90 mm petriplate containing approximately 30 ml medium
for callus induction. MS medium supplemented with different
concentrations of auxins such as 2,4-D (2,4-Dicholorophenoxy
acetic acid), NAA (Naphthalene acetic acid), IBA (Indole-3-butyric
acid) and IAA (Indole-3 acetic acid) was initially tested for callus
induction. The results were observed after 2 weeks of culture.
The best concentration of auxin was kept as standard and tried
along with different concentrations of cytokinins like BAP
(6 Benzyl aminopurine), 2iP (2-Isopentyl adenine), TDZ (Thidiazurone)
and KIN (Kinetin). Cultures were incubated at 25
2 C
under cool
fluorescent light with 16/8 h light/ dark cycle for
callus induction.