Improved Production of Galanthamine and Related Alkaloids by Methyl Jasmonate in Narcissus confusus Shoot-Clumps

Abstract

Liquid-shake cultured shoot-clumps of Narcisus confussus were treated with the commonly used biotic elicitors methyl jasmonate, arachidonic acid, chitosan and salicylic acid. The effects of these compounds on the growth of the explants, as well as on the amount of the alkaloids released to the liquid culture medium and accumulated in the tissues at the end of the experiment were studied. The obtained results showed that, in general, high doses of these compounds had a negative effect on the growth of the explants, particularly the salicylic acid. On the contrary, the addition of methyl jasmonate, mainly at 25 μM, promoted the release of galanthamine and other related alkaloids to the liquid medium in proportions of up to 300 % in relation to the control explants, and also their accumulation in tissues. The other elicitors studied did not have any interesting effects on the production of these Amaryllidaceae-type alkaloids.

Abbreviations

BA:6-benzyladenine

2,4-D:2,4-dichlorophenoxyacetic acid

DW:dry weight

FW:fresh weight

MJ:methyl jasmonate

AA:arachidonic acid

CHI:chitosan

SA:salicylic acid

FNGAL:N-formylnorgalanthamine

GAL:galanthamine

HAEM:haemanthamine

HOM:homolycorine

PTZ:pretazettine

Narcissus confusus - Amaryllidaceae - elicitors - liquid culture - methyl jasmonate - galanthamine - alkaloid production

References

• 1 Bastida J, Viladomat F. Alkaloids of Narcissus. In: Hanks G, editor Medicinal and Aromatic Plants - Industrial Profiles: The Genus Narcissus. Andover; Taylor and Francis 2002: pp 141-213
• 2 Bastida J, Viladomat F, Llabrés J M, Codina C, Feliz M, Rubiralta M. Alkaloids from Narcissus confusus .  Phytochemistry. 1987;  26 1519-24
• 3 Bergoñón S, Codina C, Bastida J, Viladomat F, Melé E. Galanthamine production in ”shoot-clump” cultures of Narcissus confusus in liquid-shake medium.  Plant Cell Tissue Org Cult. 1996;  45 191-9
• 4 Sellés M, Bergoñón S, Viladomat F, Bastida J, Codina C. Effects of sucrose on growth and galanthamine production in shoot-clumps cultures of Narcissus confusus in liquid-shake medium.  Plant Cell Tiss Org Cult. 1997;  49 129-36
• 5 Sellés M, Viladomat F, Bastida J Codina C. Callus induction, somatic embryogenesis and organogenesis in Narcissus confusus: correlation between the state of differentiation and the content of galanthamine and related alkaloids.  Plant Cell Rep. 1999;  18 646-51
• 6 Facchini P J, Johnson A G, Poupart J, De Luca V. Uncoupled defense gene expression and antimicrobial alkaloid accumulation in elicited opium poppy cell cultures.  Plant Physiol. 1996;  111 687-97
• 7 Gantet P, Imbault N, Thiersault M, Doireau P. Inhibition of alkaloid accumulation by 2,4-D in Catharanthus roseus cell suspension is overcome by methyl jasmonate.  Acta Bot Gall. 1997;  144 501-8
• 8 Zabetakis I, Edwars R, O’Hagan D. Elicitation of tropane alkaloid biosynthesis in transformed root cultures of Datura stramonium .  Phytochemistry. 1999;  50 53-6
• 9 Kim J H, Shin J H, Lee H J, Chung I S, Lee H J. Effect of chitosan on indirubin production from suspension culture of Polygonum tinctorium .  J Ferment Bioengin. 1997;  83 206-8
• 10 Spollansky T C, Pitta-Alvarez S I, Giulietti A M. Accumulation and release of tropane alkaloids in hairy root cultures of Brugmansia candida: Use of chitosan as elicitor.  In Vitro Cell Develop Biol-Plant. 1998;  34 65A
• 11 Ciddi V, Srinivasan V, Shuler M L. Elicitation of Taxus sp. cell cultures for production of taxol.  Biotechnol Lett. 1995;  17 1343-6
• 12 Shin S W, Kim Y S, Lim S. Elicitors for the regulation of baccatin III biosynthesis in plant culture system.  Yakhak Hoeji. 2000;  44 60-5
• 13 Pitta-Alvarez S I, Spollansky T C, Giulietti A M. The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmansia candida .  Enzyme Microb Tech. 2000;  26 252-8
• 14 Colque R, Viladomat F, Bastida J, Codina C. Effect of PPM on alkaloid production in Narcissus confusus shoot-clumps.  Acta Hortic. 2001;  560 267-70
• 15 López S, Bastida J, Viladomat F, Codina C. Solid-phase extraction and reversed-phase high-performance liquid chromatography of the five major alkaloids in Narcissus confusus .  Phytochem Anal. 2002;  13 311-5
• 16 Biondi S, Scaramagli S, Oksman-Caldentey K, Poli F. Secondary metabolism in root and callus cultures of Hyoscyamus muticus L.: The relationship between morphological organisation and response to methyl jasmonate.  Plant Sci. 2002;  163 563-9
• 17 Sheludko Y, Gerasimenko I, Unger M, Kostenyuk I, Stöckigt J. Induction of alkaloid diversity in hybrid plant cell cultures.  Plant Cell Rep. 1999;  18 911-8
• 18 Brodelius P, Funk C, Häner A, Villegas M. A procedure for the determination of optimal chitosan concentrations for elicitation of cultured plant cells.  Phytochemistry. 1989;  28 2651-4
• 19 Lee K T, Hirano H, Yamakawa T, Kodama T, Igarashi Y, Shimomura K. Responses of transformed root culture of Atropa belladona to salicylic acid stress.  J Biosci Bioeng. 2001;  91 586-9