LCMS and GCMS for the Screening of Alkaloids in Natural and <i>in Vitro</i> Extracts of <i>Leucojum aestivum</i>

Ptak, Agata; Tahchy, Anna El; Dupire, François; Boisbrun, Michel; Henry, Max; Chapleur, Yves; Mos, M; Laurain-Mattar, Dominique

doi:10.1021/np800585c

Cited by 48 publications

(54 citation statements)

References 34 publications

(62 reference statements)

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“…Several other compounds were also detected in in vitro extracts only, but they could not be identified. Dramatic variations in the profile of alkaloids were observed previously in tissue cultures of Leucojum aestivum, depending upon the balance of growth substances and the state of differentiation [11]. The positive effect of 2,4-D combined with BA was also shown in berberine production by in vitro culture of Coscinium fenestratum [46].…”

Section: Effect Of Auxins On Alkaloid Profile In Callus Cultures Of Lmentioning

confidence: 77%

“…Leaves were isolated from Leucojum aestivum L. 60 bulbs chilled for 12 weeks at 5ºC were surface-sterilized as previously described by Ptak et al [11]. In vivo bulbs were used for both in vitro culture establishment and chemical analysis.…”

Section: Plant Materialsmentioning

confidence: 99%

“…In the present study, the alkaloid profile of Leucojum aestivum calli was correlated with the type of auxin (2,4-D, picloram and dicamba) and its concentration (25 and 50 µM) used in the medium (Table 4). In order to study the effect of various exogenous auxins on the alkaloid profile of bulbs and in vitro callus cultures of Leucojum aestivum, capillary GC-MS was used according to the method previously described by Ptak et al [11]. Following the alkaloid extraction and purification of all samples, i.e.…”

Section: Effect Of Auxins On Alkaloid Profile In Callus Cultures Of Lmentioning

confidence: 99%

“…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].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2013

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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).

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Section: Effect Of Auxins On Alkaloid Profile In Callus Cultures Of Lmentioning

confidence: 77%

Section: Plant Materialsmentioning

confidence: 99%

Section: Effect Of Auxins On Alkaloid Profile In Callus Cultures Of Lmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2013

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“…Inhibitors of ethylene biosynthesis increased paclitaxel production in cell culture of Taxus (Tabata 2004). Our preliminary results (Ptak et al 2009) showed that the control of ethylene production can be important for somatic embryogenesis and for alkaloid synthesis in L. aestivum culture. A stimulatory effect of an ethylene inhibitor (STS) and of KMnO 4 was observed on embryogenic callus induction and alkaloid production.…”

Section: Introductionmentioning

confidence: 65%

Effects of ethylene on somatic embryogenesis and galanthamine content in Leucojum aestivum L. cultures

Ptak

Tahchy

Wyżgolik

et al. 2010

Plant Cell Tiss Organ Cult

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The influence of ethylene on in vitro morphogenesis of Leucojum aestivum and galanthamine accumulation was studied. Calli were cultivated on Murashige and Skoog (MS) medium supplemented with 25 lM 4-amino-3,5,6-trichloropicolinic acid (picloram) and 0.5 lM benzyladenine (BA). During incubation under these conditions, callus cultures produced ethylene (9.5 nL/g fresh weight: F.W.) whereas no ethylene was found in somatic embryos cultivated on medium supplemented with 0.5 lM a-naphthalene acetic acid (NAA) and 5 lM zeatin. Application of the precursor of ethylene 1-aminocyclopropane-1-carboxylic acid (ACC) increased ethylene production in both cultures, and decreased callus growth by a factor of 1.2, whereas callus growth was enhanced by a factor of 1.1 in the presence of an inhibitor of ethylene silver nitrate (AgNO 3 ) or by a factor of 1.2 with an absorbent potassium permanganate (KMnO 4 ). ACC enhanced the induction of somatic embryos and the development of globular embryos. Removal of ethylene by KMnO 4 during somatic embryogenesis led to the development of plants with greater length. Silver thiosulphate (STS) induced galanthamine production in callus cultures (0.1% dry weight), whereas ACC induced galanthamine production in somatic embryo cultures (2% dry weight).

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Galanthamine production by Leucojum aestivumL. shoot culture in a modified bubble column bioreactor with internal sections

Georgiev

Ivanov

Berkov

et al. 2012

Engineering in Life Sciences

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Shoot culture of summer snowflake (Leucojum aestivum L.) was successfully cultivated in an advanced modified glass‐column bioreactor with internal sections for production of Amaryllidaceae alkaloids. The highest amounts of dry biomass (20.8 g/L) and galanthamine (1.7 mg/L) were achieved when shoots were cultured at 22°C and 18 L/(L·h) flow rate of inlet air. At these conditions, the L. aestivum shoot culture possessed mixotrophic‐type nutrition, synthesizing the highest amounts of chlorophyll (0.24 mg/g DW (dry weight) chlorophyll A and 0.13 mg/g DW chlorophyll B). The alkaloids extract of shoot biomass showed high acetylcholinesterase inhibitory activity (IC50 = 4.6 mg). The gas chromatography–mass spectrometry (GC/MS) profiling of biosynthesized alkaloids revealed that galanthamine and related compounds were presented in higher extracellular proportions while lycorine and hemanthamine‐type compounds had higher intracellular proportions. The developed modified bubble‐column bioreactor with internal sections provided conditions ensuring the growth and galanthamine production by L. aestivum shoot culture.

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LCMS and GCMS for the Screening of Alkaloids in Natural and in Vitro Extracts of Leucojum aestivum

Cited by 48 publications

References 34 publications

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

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

Effects of ethylene on somatic embryogenesis and galanthamine content in Leucojum aestivum L. cultures

Galanthamine production by Leucojum aestivumL. shoot culture in a modified bubble column bioreactor with internal sections

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