A high-frequency, season-independent, in vitro regeneration of Ficusreligiosa was
developed, followed by comparative acetylcholinesterase inhibitory (AChEI) activity
assay of the in vitro raised and conventionally grown plants. The use of AChEI
activity is the most accepted strategy for the treatment of Alzheimer disease. Fully
expanded, mature leaves were cut into different segments to initiate the cultures.
The middle section of the leaf in vertical orientation with cut portion inserted
inside the medium was found most suitable for direct shoot regeneration. Leaf
explants responded with nearly consistent frequency (60–66.67 %) throughout the
year. To obtain high frequency response with enhanced shoot multiplication rate, 32
plant growth regulator regimes were screened amongst which benzylaminopurine at
5.0 mg/l was found most suitable, yielding 100 % response and maximum number of
shoots per explant (7.93); same concentration was also most supportive for repeated
multiplication (6.53 shoots). The quality of the shoots and multiplication rate
could be significantly enhanced (24.35 shoots) when adenine sulphate, glutamine and
phloroglucinol, in an optimised concentration, were additionally supplemented. The
clonal nature of the micropropagated plants was confirmed by random amplified
polymorphic DNA analysis. A comparative analysis of AChEI activity was carried out
amongst the methanolic extracts of stem segments of the mother plant, randomly
selected seedlings of different age (4 and 6 months old) of the same mother plant
and randomly selected micropropagated plants of different age (3 and 6 months age).
The mother plant sample showed effective AChEI activity, with
IC50 of 66.46 μg/ml while seedlings, of different age
groups, performed poorly (6-month-old seedlings, Se-16M,
yielded IC50 of 20,538.46 μg/ml, while two randomly selected
4 months’ aged seedlings, Se-24M and
Se-34M exhibited IC50 of 19,341.03
and 24,281.70 μg/ml). On the other hand, various micropropagated plants, 2 of
3 months (MiP-13M, MiP-23M) and 2 of
6 months (MiP-36M and MiP-46M) age
behaved like the mother plant, exhibiting IC50 values of
71.87, 72.91, 67.65 and 69.65 μg/ml, respectively.