Screening of Amaryllidaceae alkaloids in bulbs and tissue cultures of Narcissus papyraceus and four varieties of N. tazetta.

Narcissus spp. are an economically important crop for medicines in relation with the alkaloids production, mainly galanthamine, an acetylcholinesterase inhibitor used for the treatment of Alzheimer's disease. In this article an extensively study of the phytochemistry of both bulbs of different species and varieties of Narcissus grown in Iran and in vitro culture of these plants was investigated. In particular, the Amaryllidaceae alkaloid profile and the galanthamine and lycorine contents in wild bulbs of Narcissus papyraceus (G5) and four varieties of Narcissus tazetta (N. tazetta var. Shahla (G4), N. tazetta var. Shastpar (G1), N. tazetta var. Meskin (G2), N. tazetta var. Panjehgorbei (G3)), growing in Iran are reported. The alkaloid profiles were investigated by GC-MS and LC-MS and the quantitative analysis was performed using GC-MS. In total, thirty alkaloids were identified among them nine alkaloids were observed with the both methods of analysis. The variety Meskin of N. tazetta (G2), showed the highest diversity of alkaloids and the highest content in galanthamine. On this last species (G2) and on N. tazetta var. Shahla (G4), the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (Picloram) and naphthalene acetic acid (NAA) at concentrations of 25 and 50 μM were studied on the induction of callus and its capacity to induce organogenesis and alkaloid diversity. All auxins, at the concentrations of 25 and 50 μM, produced calli. Bulblets and roots were formed on calli grown only in the presence of 25 or 50 μM NAA. GC-MS analyses showed the presence of galanthamine and lycorine in calli, roots and bulblets, with all auxins whatever the concentration used while demethylmaritidine and tazettine were found in differentiated tissue cultures cultivated on the medium containing NAA (25 or 50 μM) or in calli initiated with Picloram (50 μM). Precursor 4'-O-methylnorbelladine (MN) of Amaryllidaceae alkaloids feeding was found to significantly improve the accumulation of both galanthamine (82 μg/g DW) and lycorine (1800 μg/g DW) in bulblets of N. tazetta var. Meskin (G2).