Effects of sucrose and plant growth regulators on acetylcholinesterase inhibitory activity of alkaloids accumulated in shoot cultures of Amaryllidaceae

Abstract

The influence of sucrose (30, 60, 90 and 120 g/L), activated charcoal (5 and 10 g/L), and various levels of several plant growth regulators (6-benzyladenine, naphthalene-1-acetic acid, 2,4-dichlorophenoxyacetic acid, and picloram) on organogenesis (bulb and root development) and the accumulation of alkaloid and galanthamine in shoot cultures of three Amaryllidaceae species (Narcissus pseudonarcissus, Galanthus elwesii, and Leucojum aestivum) was investigated in a full-factorial experiment. Alkaloid extracts were analyzed by gas chromatographymass spectrometry, leading to the quantification of galanthamine and to the identification of other alkaloids. The different extracts were then subjected to an Ellman test to evaluate the inhibitory activity of acetylcholinesterase. The highest contents of galanthamine [0.02-0.1% dry weight (DW) depending on the plant species] were always accompanied with a high level of acetylcholinesterase inhibition ([30%). However, some samples containing low amounts of galanthamine (0.005% DW) showed high inhibitory activities (40-80%). These findings demonstrate the presence of Amaryllidaceae alkaloids that have not yet been identified as having anti-acetylcholinesterase activity.

Accumulation of hydroxybenzoic acids and other biologically active phenolic acids in shoot and callus cultures of Aronia melanocarpa (Michx.) Elliott (black chokeberry)

“…anthraquinones (hypericin, pseudohypericin), phloroglucine derivate (hyperforin) in Hypericum sp. shoot cultures (Coste et al 2011), flavonoid (rutin) in Morus alba in vitro cultures (Lee et al 2011), terpenoids in Camellia sinensis suspension cultures (Grover et al 2012), and alkaloid (galanthamine) production in in vitro cultures of three Amaryllidaceae species (El Tahchy et al 2011). Studies performed in our laboratory have confirmed the effects of NAA and BA concentrations in L-S medium on free phenolic acids accumulation in in vitro cultures of R. graveolens (Ekiert et al 2009) and its subspecies R. graveolens ssp.…”

Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus

“…The low Amaryllidaceae alkaloid accumulation commonly observed in in vitro cultures of Leucojum aestivum [6,[9][10][11][12][13] was probably due to the fact that the level of the alkaloid biosynthesis is strongly influenced by cellular differentiation [10,[14][15][16]. In particular, enhanced synthesis of galanthamine accompanied development of somatic embryos [17].…”

Production and antioxidant capacity of tissue cultures from four Amaryllidaceae species

“…The present study demonstrates that a micropropagation system, not involving the production of embryogenic calli, can also be established for this species. Besides our report, several studies reported the production of L. aestivum tissue cultures [7,26], but none of them involved the assay of antioxidant capacities (see below). All cultures showed high shoot / whole plant and (except L. aestivum) high bulblet regeneration frequencies (Tab.…”

“…Several tissue cultures of Amaryllidaceae involve the growth regulators NAA, IAA as auxins and BA as a cytokinin [7,10,27,28]. Auxin/cytokinin combinations involving growth regulators mentioned are potent inducers of embryogenic callus formation and/or mass plant propagation in this family.…”

“…Particularly, there is no need to harvest plants from their natural habitats for the production of biologically active compounds, because tissue culture procedures could supply raw material for such purposes. Some important examples are Galanthus elwesii, Leucojum aestivum, and Narcissus pseudonarcissus [7,8].…”