Micropropagation of Ficus religiosa L. via leaf explants and comparativeevaluation of acetylcholinesterase inhibitory activity in the micropropagated andconventionally grown plants

摘要

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 outamongst the methanolic extracts of stem segments of the mother plant, randomlyselected seedlings of different age (4 and 6 months old) of the same mother plantand randomly selected micropropagated plants of different age (3 and 6 months age).The mother plant sample showed effective AChEI activity, withIC50 of 66.46 μg/ml while seedlings, of different agegroups, performed poorly (6-month-old seedlings, Se-16M,yielded IC50 of 20,538.46 μg/ml, while two randomly selected4 months’ aged seedlings, Se-24M andSe-34M exhibited IC50 of 19,341.03and 24,281.70 μg/ml). On the other hand, various micropropagated plants, 2 of3 months (MiP-13M, MiP-23M) and 2 of6 months (MiP-36M and MiP-46M) agebehaved like the mother plant, exhibiting IC50 values of71.87, 72.91, 67.65 and 69.65 μg/ml, respectively.