2.2. Establishment of callus cultures
When the seedling reached 30 days old, leaves (about 0.25 cm
in length) of all genotypes were excised from different S.
marianum seedlings and then cultured on MS-basal-medium
supplemented with 0.25 mg L1 2,4-D + 0.25 mgL1 Kin.
Cultures were incubated in a growth chamber for one month
for callus initiation. Calli cultures were subcultured monthly
for 6 months on that medium for increasing of calli tissues
for further experiments.