RESUMEN
The limited production of bioactive essential oils in natural plants does not meet the increasing worldwide market demand. Plant cell culture technology can be used for the higher production of industrially important essential oils. In the present study, a suitable method for production of essential oils was developed through establishment and elicitation of adventitious roots (AR) in a medicinally important plant Artemisia amygdalina D. The results indicated that leaf explants cultured on solid Murashige and Skoog (MS) media supplemented with 1.0 mg/L α- naphthalene acetic acid (NAA) and 4% sucrose instigated the higher AR induction frequency (90 ± 4.25) and maximum AR biomass (fresh biomass: 17.7 g/L). Furthermore, in the AR when transiently elicited with different elicitors for different time periods, methyl jasmonate (Me-J: 0.5 mg/L) resulted in the higher production of total phenolic content (TPC: 3.6 mg), total flavonoid content (TFC: 2.3 mg) and phenylalanine ammonia-lyase (PAL: 4.8 U/g×FW) activity, respectively. Nonetheless, considerable levels of the major bioactive compounds such as α-thujene (6.8%), α-pinene (8.3%), 1,8-cineole (16.2%), camphor (8.4%) and verbenole (10.2%) were recorded in the Me-J treated AR. Thus, a feasible protocol for production of essential oils through AR in A. amygdalina was established, which can be exploited for commercial production of the industrially important terpenes.
RESUMEN
Clinically, available synthetic chemotherapeutics in the treatment for leishmaniasis are associated with serious complications, such as toxicity and emergence of resistance. Natural products from plants can provide better remedies against the Leishmania parasite and can possibly minimize the associated side effects. In this study, various extracts of the callus cultures of Artimisia scoparia established in response to different plant growth regulators (PGRs) were evaluated for their anti-leishmanial effects against Leishmania tropica promastigotes, followed by an investigation of the possible mechanism of action through reactive apoptosis assay using fluorescent microscopy. Amongst the different callus extracts, higher anti-leishmanial activity (IC50:19.13 µg/mL) was observed in the callus raised in-vitro in the presence of 6-Benzylaminopurine (BA) plus 2,4-Dichlorophenoxyacetic Acid (2,4-D) at the concentration of 1.5 mg/L, each. Further, the results of apoptosis assay showed a large number of early-stage apoptotic (EA) and late-stage apoptotic (LA) cells in the Leishmania under the effect of callus extract grown in-vitro at BA plus 2,4-D. For the determination of the potent natural products in the callus extracts responsible for the anti-leishmanial activity, extracts were subjected to Gas chromatography-mass spectrometry (GC-MS) for the metabolite analysis. Nonetheless, higher levels of the metabolites, such as nerolidol (22%), pelletierine (18%), aspidin (15%) and ascaridole (11%) were detected in the callus grown in vitro at BA plus 2,4-D (1.5 mg/L, each). This protocol determines a novel method of production of anti-leishmanial natural products through callus cultures of A. scoparia, a medicinal plant.