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

Abstract

In vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

Chemical Compositions, Somatic Embryogenesis, and Somaclonal Variation in Cumin

“…Embryos formed in the presence of phenolic compositions lack sufficient amounts of sucrose to carry out the metabolic activity and thus stop in the globular stage. It should be noted that the inhibitory effect of phenolic compositions due to their high concentration in cells leads to protein deposition, enzyme deactivation, or membrane damage [18, 64]. However, in some studies, it has been reported that phenols usually have a reinforcing effect on somatic embryogenesis, but it is possible that phenols interaction with alcohols and other phytochemical compositions of essential oil inhibits their reinforcing effect on embryogenesis.…”

Induction of somatic embryogenesis and biochemical characterization of Cordyline australis (G. Forst.) Endl. ‘Red Star’ callus

“…minor (Skrzypek et al, 2007) and Galanthus elwesii (Bach et al, 2004). In contrast, Hrubcová et al (1994), Nieves et al (2003) and Ptak et al (2013) reported that non-embryogenic calli produced more phenolics than embryogenic ones. The study of Ozyigit et al (2007) indicates that the role of the phenolic compounds depends on its chemical structure, plant species, the biological process studied (organogenesis or somatic embryogenesis), and its developmental stage.…”

“…Similar trends were also observed in studies on somatic embryogenesis in the cultures of Saccharum sp. (Garcia et al, 2007), Narcisus (Malik et al, 2009), and Leucojum aestivum (Ptak et al, 2013).…”

Leucojum aestivum L. in vitro bulbs induction and acclimatization

“…In vitro plants of Leucojum aestivum were regenerated from somatic embryos obtained on a solid Murashige and Skoog (MS) [13] medium supplemented with 5 µM zeatin and 0.5 µM benzyladenine (BA) according to the procedure described by Ptak et al [8,12]. For experiment 28-day-old plants (0.3-0.5 g FW each plant), were transferred to MS medium contained 30 g L -1 sucrose and 10 µM paclobutrazol, or 10 µM ancymidol, or 10 µM GA 3 .…”

“…Micropropagation ensures rapid multiplication of selected genotypes, allowing the useful metabolites to be collected in greater quantities. Several studies on Leucojum aestivum multiplication by organogenesis and somatic embryogenesis have been performed [5,8,9]. Species that naturally produce bulbs, like Leucojum aestivum can be induced to form bulbs in in vitro culture.…”