Elicitation – a tool to improve secondary metabolites production in Melissa Officinalis L. Suspension cultures / Elicitácia ako nástroj na zlepšenie produkcie sekundárnych metabolitov v suspenzných kultúrach Melissa Officinalis L.

J. Urdová 1 , M. Rexová 2 , P. Mučaji 1 , and A. Balažová 3
  • 1 Comenius University in Bratislava, Faculty of Pharmacy, Department of Pharmacognosy and Botany, Slovak Republic
  • 2 Comenius University in Bratislava, Faculty of Pharmacy, Department of Pharmacology and Toxicology, Slovak Republic
  • 3 Comenius University in Bratislava, Faculty of Pharmacy, Department of Cell and Molecular Biology of Drugs, Slovak Republic


Our recent study is focussed on the investigation of the influence of biotic (Botrytis cinerea, methyl jasmonate) and abiotic (cadmium and cobalt chloride) elicitors on Melissa officinalis L. (lemon balm) suspension cultures production properties. The plant material was treated with different concentrations of methyl jasmonate (10 and 100 μmol.l−1), with a phytopathogenic fungus hydrolyzate (Botrytis cinerea, 1 and 2 ml with glucose equivalent 24 μg.ml−1), cadmium and cobalt (II) chloride (both at concentrations 100 μmol.l−1 and 1 μmol.l−1). Elicited suspension cultures were harvested after 24, 48 and 72 h of elicitor treatment. The elicitation effect was evaluated based on hydroxycinnamic acid derivatives content expressed as rosmarinic acid in dose- and time-dependent manners. The Botrytis cinerea hydrolyzate exhibited the best effect on Melissa officinalis L. suspension cultures production properties among biotic elicitors (glucose equivalent 24 mg.ml−1, 1 ml, after 72 h of treatment). The hydroxycinnamic acid derivatives content increased in triplicate compared to non-elicited samples. Methyl jasmonate affected the suspension cultures production properties moderately at concentration 10 μmol.l−1. Chlorides salts of cobalt and cadmium stimulated phenolic compounds production effectively at concentrations 100 μmol.l−1. The presence of cobalt and cadmium ions in suspension cultures growth media increased the hydroxycinnamic acid derivatives content twofold and quadruple, respectively.

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  • [1] Arnow LE. Colorimetric determination of the components of 3,4-dihydroxyphenylalaninetyrosine mixtures. J Biol Chem. 1937;118:531-537.

  • [2] Balážová A, Blanárikova V, Bilka F, Bilková A, Kiňová Sepová H. Efekt troch rôznych elicitorov na produkciu sanguinarínu suspenznými kultúrami nízko-morfínovej odrody maku siateho (Papaver somniferum L.). Čes Slov Farm. 2011;60:237-240.

  • [3] Balážová A, Blanárikova V, Bilka F, Bilková A. Effect of combined biotic and abiotic elicitor on the sanguinarine formation in cell suspension cultures of Eschscholtzia californica Cham. Acta Facult Pharm Univ Comenianae. 2008;55:58-63.

  • [4] Bhuvaneswari CH, Rao K, Gandi S, Giri A. Abiotic elicitation of gymnemic acid in the suspension cultures of Gymnema sylvestre. World J Microbiol and Biotechnol. 2012; 28:741-747.

  • [5] Bilka F, Balažová A, Bilková A, Holková I. Porovnanie produkcie sanguinarínu suspenznými kultúrami rastlín čeľade Papaveraceae. Čes Slov Farm. 2012;61:267-270.

  • [6] Bilková A, Bilka F, Blanáriková V, Bezáková L. Effect of excess of cupric sulfate on sanguinarine formation and activities of amine oxidase and polyphenol oxidase in cell suspension cultures of Papaver somniferum. Biologia. 2006;60:661-664.

  • [7] Cai Z, Kastell A, Speiser C, Smetanska I. Enhanced resveratrol production in Vitis vinifera cell suspension cultures by heavy metals whitout loss of cell viability. App Biochem and Biotechnol. 2013;171:330-340.

  • [8] European Pharmacopoeia. 8th edition. 2014; 1369-1370 (01/2013:1560, Rosmarini folium).

  • [9] Fialová S, Tekeľová D, Mrlianová M, Grančai D. The determination of phenolics compounds and antioxidant activity of mints and balms cultivated in Slovakia. Acta Facult Pharm Univ Comenianae. 2008;55:96-102.

  • [10] Holková I, Bezáková L, Bilka F, Balážová A, Vanko M, Blanárikova V. Involvement of lipoxygenase in elicitor-stimulated sanguinarine accumulation in Papaver somniferum suspension cultures. Plant Phys Biochem. 2010;48:887-892.

  • [11] Ketchum REB, Gibson DM, Croteau RB, Shuler ML. The kinetics of taxol accumulation in cell suspension cultures of Taxus following elicitation with methyl jasmonate. Biotech Bioeng.1999; 62:97-105.

  • [12] Lee-Parson CWT, Erturk S, Tengtrakool J. Enhancement of ajmalicine production in Catharanthus roseus cellcultures with methyl jasmonate is dependent on timing and dosage of elicitation. Biotechnol Lett. 2004;26(20):1595-1599.

  • [13] López-Millán AF, Sagardoy R, Solanas M, Abadía A, Abadía J. Cadmium toxicity in tomato Lycopersicon esculentum) plants grown in hydroponics. Env Exp Bot. 2009;65:379-385.

  • [14] Marongui B, Porcedda S, Piras A, Rosa A, Deiana M, Dessi MA. Antioxidant activity of supercritical extract of Melissa officinalis subsp. officinalis and Melissa officinalis subsp. inodora. Phytotherapy Research. 2004;18:789-792.

  • [15] Mizukami H, Tabira Y, Ellis BE. Methyl jasmonate-induced rosmarinic acid biosynthesis in Lithospermum erythrorhizon cell suspension cultures. Plant Cell Rep. 1993;12:706-709.

  • [16] Moscatiello R, Baldan B, Navazio L. Plant cell suspension cultures. Methods Mol Biol. 2013;953:77-93.

  • [17] Mrlianová M, Tekeľová D, Felklová M, Tóth J, Musil P, Grančai D. Comparison of the quality of Melissa officinalis L. cv. Citra with melissas of Europeam origin. Čes Slov Farm. 2001;50:299-302. (In Slovak.)

  • [18] Mrlianová M, Tekeľová D, Felklová M, Reinöhl V, Tótj, J. The influence of the harvest cut height on the quality of the herbal drugs Melissae folium and Melissae herba. Planta Med. 2002;68:178-180.

  • [19] Mrlianová M, Tekeľová D, Tóth J, Grančai D. The content of essential oil and total hydroxycinnamic derivatives in leaves of Melissa officinalis L. of domestic and foreign origin. Farm Obzor. 2011;80:176-180. (In Slovak.)

  • [20] Namdeo A. Plant Cell Elicitation for Production of Secondary Metabolites. Pharmacogn Rev. 2007;1:69-79.

  • [21] Oksman-Caldentey KM, Inzé D. Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. Trends Plant Sci. 2004;9:433-440.

  • [22] Ram M, Prasad VK, Singh SK, Hada BS, Kumar S. Influence of salicylic acid and methyl jasmonate elicitation on anthocyanin production in callus cultures of Rosa hybrida L. Plant Cell Tiss and Organ Cult. 2013; 113:459-467.

  • [23] Ramakrishna A, Ravishankar GA. Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav. 2011; 6:1720-1731.

  • [24] Srivastava NK, Srivastava AK. Influence of some heavy metals on growth, alkaloid content and composition in Catharanthus roseus L. Indian J Pharm Sci. 2010; 72(6):775-778.

  • [25] Tóth J, Mrlianová M, Tekeľová D, Koreňová M. Rosmarinic acid - an important phenolic active compound of lemon balm (Melissa officinalis L.). Acta Facult Pharm Univ Comenianae. 2003; 50:139-146.


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