Micropropagation of Ficus religiosa L. via leaf explants and comparative evaluation of acetylcholinesterase inhibitory activity in the micropropagated and conventionally grown plants

Abstract

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.

In Vitro Adventitious Shoot Regeneration through Direct and Indirect Organogenesis from Seedling-derived Hypocotyl Segments of Ficus religiosa L.: An Important Medicinal Plant

“…Many studies proved the positive influence of cytokinin on cell division and shoot regeneration (Arab et al, 2014;Jafari et al, 2017;Gill, 2011, 2014). Hesami et al (2017a) and Siwach and Gill (2014) reported that TDZ is more effective than BAP on the number of shoot regeneration, whereas various studies take a radical point of view and alleged that BAP is more effective than TDZ (Deshpande et al, 1998;Hassan et al, 2009;. Therefore, these conflicting results might be related to the use of various explants and effects of various genotypes.…”

“…In another study, reported that the maximum number of multiple shoots from nodal segments of F. religiosa was achieved on WPM containing 1.0 mg · L -1 BAP along with 0.5 mg · L -1 IAA, and these findings confirmed our results. The influence of cytokinins on micropropagation can be varied based on the kind of culture medium, the variety of plants, and the age of explants (Bhojwani and Dantu, 2013;Hesami et al, 2017b;Siwach and Gill, 2014).…”

“…indicated that the highest budbreak frequency (100%) via mature nodal segments was obtained on Woody Plant Medium (WPM) supplemented with 1.0 mg · L -1 BAP along with 0.5 mg · L -1 IAA. Also, Siwach and Gill (2014) reported that the highest regeneration frequency (100%) via mature leaf segments was reached in MS medium supplemented with 5.0 mg · L -1 BAP. According to the mentioned studies, it became clear that the response of mature explants and the contamination frequency could be influenced by the season of explants collection, restricting the culture initiation experiment to a particular time period of the year .…”

“…Mostly, the characterization and isolation of metabolites in this plant are carried out from naturally occurring sources. Such sources are time-and labor consuming and are influenced by environmental conditions and high religious values (Siwach and Gill, 2014). Hence, commercial processing demands substitute sources (Singh et al, 2011).…”

Effects of thidiazuron on in vitro shoot regeneration of Morus alba

“…and Hesami et al (2017c) reported that 2 mg/l IBA and 0.1 mg/l NAA were suitable for inducing the rooting of F. religiosa. Siwach and Gill (2011) and Siwach and Gill (2014) also reported that IBA was more appropriate than NAA and IAA in inducing the rooting process in F. religiosa. IBA is more stable and less sensitive to degradation and is slowly metabolized by the peroxidase enzyme; this can be one of the possible causes for better effect of IBA in root formation as compared to other auxins (Dobránszki and da Silva, 2010).…”