Phenolic Profiles, Antimicrobial and Cytotoxic Properties of Both Micropropagated and Naturally Growing Plantlets of Calamintha sylvatica subsp. sylvatica Bromf.


  • Mustafa CÜCE Giresun University, Şebinkarahisar School of Applied Sciences, Department of Food Technology, 28400, Giresun (TR)
  • Tuba BEKİRCAN Eurasia University, Faculty of Science and Art, Department of Biochemistry, 61080, Trabzon (TR)
  • Abdul Hafeez LAGHARI Karadeniz Technical University, Faculty of Science, Department of Chemistry, 61080, Trabzon (TR)
  • Münevver SÖKMEN Karadeniz Technical University, Faculty of Science, Department of Chemistry, 61080, Trabzon; King Saud University, College of Science, Department of Zoology, 11451, Riyadh (SA)
  • Atalay SÖKMEN Konya Food and Agriculture University, Faculty of Agriculture and Natural Sciences, Department of Plant Production & Technologies, 42080, Konya; King Saud University, College of Science, Department of Zoology, 11451, Riyadh (SA)
  • Evren ÖNAY UÇAR İstanbul University, Faculty of Science, Department of Molecular Biology and Genetic, 34134 İstanbul (TR)
  • Ali Osman KILIÇ Karadeniz Technical University, Faculty of Medicine, Department of Medical Microbiology, 61080 Trabzon (TR)



antimicrobial activity; Calamintha sylvatica; cytotoxic activity; micropropagation; rosmarinic acid


A rapid micropropagation protocol was designed to produce Calamintha sylvatica plantlets by using nodal segments as explants for the shoot formation. 6-BA favored the highest shoot formation and biomass yield, whilst kinetin was found superior for the highest shoot length (38.97 ± 2.85 mm) and node numbers (2.89 ± 0.63). Rosmarinic acid was detected as major phenolic acid, ranging from 7.59 mg/100 g to 81.44 mg/100 g. Hexane extracts from natural and in vitro propagated plantlets showed activity only against Staphylococcus aureus ATCC 25923 with MIC values at 6.25 and 3.33 m/mL, respectively while in the latter case, extracts from natural plantlets exerted higher cytotoxic activity than those of micropropagated ones (IC50 values were 83 µg/mL and 98 µg/mL on HeLa cells, respectively). C. sylvatica showed high micropropagation performance and produced remarkable amount of rosmarinic acid in vitro as well as antimicrobial and cytotoxic effect.


Alan S, Kürkçüoğlu M, Ozek T, Can Başer KH (2009). Composition of the essential oils of Calamintha tauricola P.H. Davis. Journal of Essential Oil Research 21(2):143-145.

Araniti F, Lupini A, Mercati F, Statti G, Abenavoli M (2013). Calamintha nepeta L. (Savi) as source of phytotoxic compounds: bio-guided fractionation in identifying biological active molecules. Acta Physiologiae Plantarum 35(6):1979-1988.

Arikat NA, Jawad FM, Karam NS, Shibli RA (2004). Micropropagation and accumulation of essential oils in wild sage (Salvia fruticosa Mill.). Scientia Horticulturae-Amsterdam 100(1-4):193-202.

Astani A, Reichling J, Schnitzler P (2012). Melissa officinalis extract inhibits attachment of herpes simplex virus in vitro. Chemotherapy 58(1):70-77.

Avato P, Fortunato IM, Ruta C, D’Eli R (2005). Glandular hairs and essential oils in micropropagated plants of Salvia officinalis L. Plant Science 169(1):29-36.

Baytop A (1983). Farmasötik botanic. İstanbul Üniversitesi Eczacılık Fakültesi Yayınları 36:282-285 [Pharmaceutical botanic. Istanbul University Faculty of Pharmacy Publications 36:282-285].

Baytop A (1997). Türkçe Bitki Adları Sözlügü. Atatürk Kültür, Dil ve Tarih Yüksek Kurumu. Türk Dili Kurumu Yayınları, 578, Ankara pp 512 [Turkish Plant Names Dictionary. Atatürk Culture, Language and History Supreme Council. Turkish Language Institution Publications, 578, Ankara pp 512].

Baytop T (1999). Türkiye’de Bitkilerle Tedavi. İstanbul Üniversitesi yayınları No. 3255, Eczacılık Fakültesi yayınları No. 40, İstanbul pp 304-371 [Treatment with plant in Turkey. Istanbul University publications No. No. 3255, Faculty of Pharmacy publications Nos. 40, Istanbul pp 304-371].

Boonyarikpunchai W, Sukrong S, Towiwat P (2014). Antinociceptive and anti-inflammatory effects of rosmarinic acid isolated from Thunbergia laurifolia Lindl. Pharmacology Biochemistry and Behavior 124:67-73.

Bown D (1995). The Royal Horticultural Society Encyclopedia of Herbs and Their Uses. Dorling Kindersley, New York pp 97-252.

Clinical and Laboratory Standards Institute (2013). Performance standards for antimicrobial susceptibility testing; twenty-third informational supplements. CLSI document M100-S23, Wayne, PA/USA.

Cunningham AB (1993). African medicinal plants: setting priorities at the interface between conservation and primary health care. People and Plants working paper I. Paris. UNESCO.

Cüce M, Bekircan T, Laghari AH, Sökmen M, Sökmen A, Uçar EÖ, Kılıç AO (2017). Antioxidant phenolic constituents, antimicrobial and cytotoxic properties of Stachys annua L. from both natural resources and micropropagated plantlets. Indian Journal of Traditional Knowledge 16(3):407-413.

De Silva T (1997). Industrial utilization of medicinal plants in developing countries. In: Bodeder G, Bhat KKS, Burley J, Vantomme P (Eds). Medicinal plants forest conservation and healthcare. Non-Wood Forest Products No 11, FAO, Rome, Italy.

Debnath M, Malik C, Bisen P (2006). Micropropagation: a tool for the production of high quality plant-based medicines. Current Pharmaceutical Biotechnology 7(1):33-49.

Dobravalskytė D, Venskutonis PR, Talou T (2012). Antioxidant properties and essential oil composition of Calamintha grandiflora L. Food Chemistry 135(3):1539-1546.

Döring AS, Petersen M (2014). Production of caffeic, chlorogenic and rosmarinic acids in plants and suspension cultures of Glechoma hederacea. Phytochemistry Letters 10:111-117.

European Committee on Antimicrobial Susceptibility Testing (2014). EUCAST disk diffusion test manual, version 4.0. Retrieved 2016 November 12 from

Fattahi M, Nazeri V, Torras-Claveria L, Sefidkon F, Cusido RM, Zamani Z, Palazon J (2013). A new biotechnological source of rosmarinic acid and surface flavonoids: Hairy root cultures of Dracocephalum kotschyi Boiss. Industrial Crops and Products 50:256-263.

Formisanoa C, Olivieroa F, Riganoa D, Saabb AM, Senatorea F (2014). Chemical composition of essential oils and in vitro antioxidant properties of extracts and essential oils of Calamintha origanifolia and Micromeria myrtifolia, two Lamiaceae from the Lebanon flora. Industrial Crops and Products 62:405-411.

Gamborg OL, Miller RA, Ojima K (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research 50(1):151-158.

Grigoriadou K, Maloupa E (2008). Micropropagation and salt tolerance of in vitro grown Crithmum maritimum L. Plant Cell, Tissue and Organ Culture 94(2):209-217.

Kim K-H, Tsao R, Yang R, Cui SW (2006). Phenolic acid profiles and antioxidant activities of wheat bran extracts and the effect of hydrolysis conditions. Food Chemistry 95(3):466-473.

Louvel S, Moodley N, Seibert I, Steenkamp P, Nthambeleni R, Vidal V, … Klimkait T (2013). Identification of compounds from the plant species Alepidea amatymbica active against HIV. South African Journal of Botany 86:9-14.

Lucchesini M, Mensuali-Sodi A (2010). Plant tissue culture - An opportunity for the production of nutraceuticals. Advances in Experimental Medicine and Biology 698:185-202.

Luis JC, González-Padrón MY, Pérez RM, Viera IF, González FV (2013). Rosmarinic acid: biological, pharmological and in vitro plant cell culture approximation. In: Brahmachari G (Ed). Chemistry and pharmacology of naturally occurring bioactive compounds, CRC Press, pp 471-482.

Ma Y-Q, Chen J-C, Liu D-H, Ye X-Q (2009). Simultaneous extraction of phenolic compounds of citrus peel extracts: Effect of ultrasound. Ultrasonics Sonochemistry 16(1):57-62.

Malik RA, Gupta RC, Singh V, Bala S, Kumari S (2016). New chromosome reports in Lamiaceae of Kashmir (Northwest Himalaya), India. Protoplasma 254(2):971-985.

Mohanty SK, Malappa K, Godavarthi A, Subbanarasiman B, Maniyam A (2014). Evaluation of antioxidant, in vitro cytotoxicity of micropropagated and naturally grown plants of Leptadenia reticulate (Retz.) Wight & Arn.-an endangered medicinal plant. Asian Pacific Journal of Tropical Medicine 7:S267-S271.

Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65(1-2):55-63.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3)473-497.

Oluk EA, Çakır A (2009). Micropropagation of Origanum sipyleum L., an endemic medicinal herb of Turkey. African Journal of Biotechnology 8(21):5769-5772.

Önay-Uçar E, Erol Ö, Kandemir B, Mertoğlu E, Karagöz A, Arda N (2012). Viscum album L. extracts protects HeLa cells against nuclear and mitochondrial DNA damage. Evidence-Based Complementary and Alternative Medicine 958740.

Parnham MJ, Kesselring K (1985). Rosmarinic acid. Drugs Future 10:756-757.

Pistelli L, Giovannini A, Ruffoni B, Bertoli A, Pistelli L (2010). Hairy root cultures for secondary metabolites production. In: Bio-Farms for Nutraceuticals. Springer, Boston, MA pp 167-184.

Pistelli L, Noccioli C, D'Angiolillo F, Pistelli L (2013). Composition of volatile in micropropagated and field grown aromatic plants from Tuscany Islands. Acta Biochimica Polonica 60(1):43-50.

Proestos C, Chorianopoulos N, Nychas GJE, Komaitis M (2005). RP-HPLC analysis of the phenolic compounds of plant extracts. Investigation of their antioxidant capacity and antimicrobial activity. Journal of Agricultural and Food Chemistry 53(4):1190-1195.

Ruffoni B, Mascarello C (2009). Tecniche per la propagazione in vitro degli arbusti mediterranei. Flortecnica 6:51-52.

Ruffoni B, Pistelli L, Bertoli A, Pistelli L (2010). Plant cell cultures: bioreactors for industrial production. Advances in Experimental Medicine and Biology 698:203-221.

Scarpa G, Milia M, Satta M (2000). The influence of growth regulators on proliferation and rooting of in vitro propagated myrtle. Plant Cell, Tissue and Organ Culture 62(3):175-179.

Wüst Zibetti A, Aydi A, Arauco Livia M, Bolzan A, Barth D (2013). Solvent extraction and purification of rosmarinic acid from supercritical fluid extraction fractionation waste: economic evaluation and scale-up. Journal of Supercritical Fluids 83:133-145.

Zhu F, Asada T, Sato A, Koi Y, Nishiwaki H, Tamura H (2014). Rosmarinic acid extract for antioxidant, antiallergic, and α-Glucosidase inhibitory activities, isolated by supramolecular technique and solvent extraction from Perilla leaves. Journal of Agricultural and Food Chemistry 62(4):885-892.

Ziaková A, Brandšteterová E (2003). Validation of HPLC determination of phenolic acids present in some Lamiaceae family plants. Journal of Liquid Chromatography and Related Technologies 26(3):443-453.




How to Cite

CÜCE, M., BEKİRCAN, T., LAGHARI, A. H., SÖKMEN, M., SÖKMEN, A., ÖNAY UÇAR, E., & KILIÇ, A. O. (2019). Phenolic Profiles, Antimicrobial and Cytotoxic Properties of Both Micropropagated and Naturally Growing Plantlets of Calamintha sylvatica subsp. sylvatica Bromf. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(4), 1145–1152.



Research Articles
DOI: 10.15835/nbha47411474