Origanum vulgare L. Cuttings Rhizogenesis in Microclonal Reproduction in Vitro





Oregano, Culture in vitro, Nutrient media, Cutting, Root system, Adaptation in soil


Background. Oregano (Origanum vulgare L.) is a valuable essential oil culture used in the pharmaceutical and food branches of industry as a source of biologically active substances. The optimization of microclonal propagation technology in vitro of this culture is considered topical for the development of the selection of oregano with a high content of essential oil and rapid multiplication of high-performance samples.

Objective. This article aims to study the conditions of cuttings' rooting of oregano samples, valuable in terms of breeding, obtained at the last stage of in vitro cutting.

Methods. Cuttings of eleven oregano genotypes obtained after the fourth cutting of the original maternal shoots were used as a material for the study of rhizogenesis in vitro. Roots formation and development of newly formed shoots were investigated by alteration of the carbohydrate and growth regulators composition in nutrient media for rhizogenesis. ½MS medium without growth regulators was used as a control medium. Experimental in growth regulator composition were MS-based media supplemented with one of the following components: indolylacetic acid (IAA) (2 mg/l), IAA (0.5 mg/l), indolylbutyric acid (IBA) (1 mg/l), IBA (0.5 mg/l), 0.5 mg/l IAA + 0.5 mg/l IBA. Sucrose (20 g/l) or glucose (20 g/l) was used as a carbon source. The influence of the composition of rhizogenesis media was estimated on the length of cuttings' root system, the length of newly formed shoots and the survival of regenerated plants after transfer to soil.

Results. Formation of the oregano root system and newly formed shoots at a frequency of 100% occurred for all the studied genotypes and versions of media while the survival of regenerated plants in soil as a whole in the experiment for use of sucrose as a source of carbon was 63.82 ± 11.83% and for glucose 66.45 ± 11.62%. However, depending on the genotype and composition of the nutrient media, there was a variation in root system length, length of newly formed shoots and in the survival of regenerated plants in soil. A comparison of the results of rooting with sucrose or glucose in the medium for rhizogenesis did not reveal a significant difference between these compounds, but there was a tendency for better growth of the root system when using sucrose. A significant positive correlation was established between the cuttings' root system length and the survival of regenerated plants after transfer to soil (for sucrose r = 0.57, for glucose r = 0.51, r0.05 = 0.24) and no correlation was proved between the newly formed shoots length and the survival of regenerated plants in soil (for sucrose r = –0.21, for glucose r = –0.03, r0.05 = 0.24).

Conclusions. It is proved that the influence of carbohydrate composition and type and concentration of growth regulators in the nutrient medium on the oregano cuttings' rhizogenesis obtained during the last cycle of microclonal propagation in vitro is genotype-specific. ½MS medium supplemented with 2 mg/l of indolylacetic acid, with sucrose (20 g/l) as a carbon source, proved to be most effective for root development in vitro and cuttings' adaptation in the soil for most of the genotypes studied.


Skoufogianni E, Solomou A, Danalatos N. Ecology, cultivation and utilization of the aromatic greek oregano (Origanum vulgare L.): A review. Not Bot Horti Agrobo. 2019;47(3):545-52. DOI: 10.15835/nbha47311296

Spyridopoulou K, Fitsiou E, Bouloukosta E, Tiptiri-Kourpeti A, Vamvakias M, Oreopoulou A, et al. Extraction, chemical com­position, and anticancer potential of Origanum onites L. essential oil. Molecules. 2019;24(14):2612. DOI: 10.3390/molecules24142612

Ramadan M, Elbanna K. The oil of oregano (Origanum vulgare). INFORM International News on Fats, Oils, and Related Materials. 2017;28(3):18-20. DOI: 10.21748/inform.03.2017.18

Kosakowska O, Czupa W. Morphological and chemical variability of common oregano (Origanum vulgare L. subsp. vulgare) occurring in eastern Poland. Herba Polonica. 2018;64(1):11-21. DOI: 10.2478/hepo-2018-0001

George EF, Hall MA, Klerk GJD. Micropropagation: uses and methods. In: George EF, Hall MA, Klerk GJD, editors. Plant propagation by tissue culture. Dordrecht: Springer; 2008. p. 29-64. DOI: 10.1007/978-1-4020-5005-3_2

Menezes N, Martins W, Longhi D, de Aragão G. Modeling the effect of oregano essential oil on shelf-life extension of vacuum-packed cooked sliced ham. Meat Sci. 2018;139:113-9. DOI: 10.1016/j.meatsci.2018.01.017

Parreira D, Alcántara-de la Cruz R, Leite G, Ramalho F, Zanuncio J, Serrão J. Quantifying the harmful potential of ten essen­tial oils on immature Trichogramma pretiosum stages. Chemosphere. 2018;199:670-5. DOI: 10.1016/j.chemosphere.2018.02.083

Khosravi A, Sharifzadeh A, Nikaein D, Almaie Z, Gandomi Nasrabadi H. Chemical composition, antioxidant activity and anti­fungal effects of five Iranian essential oils against Candida strains isolated from urine samples. J Mycol Med. 2018;28(2):355-60. DOI: 10.1016/j.mycmed.2018.01.005

Elshafie H, Armentano M, Carmosino M, Bufo S, de Feo V, Camele I. Cytotoxic Activity of Origanum vulgare L. on hepa­tocellular carcinoma cell line HepG2 and evaluation of its biological activity. Molecules. 2017;22(9):1435. DOI: 10.3390/molecules22091435

Olijhoek D, Hellwing A, Grevsen K, Haveman L, Chowdhury M, Løvendahl P, et al. Effect of dried oregano (Origanum vulgare L.) plant material in feed on methane production, rumen fermentation, nutrient digestibility, and milk fatty acid composition in dairy cows. J Dairy Sci. 2019;102(11):9902-18. DOI: 10.3168/jds.2019-16329

Prasanna R, Ashraf E, Essam M. Chamomile and oregano extracts synergistically exhibit antihyperglycemic, antihyper­lipidemic, and renal protective effects in alloxan-induced diabetic rats. Can J Physiol Pharmacol. 2017;95(1):84-92. DOI: 10.1139/cjpp-2016-0189

Han X, Parker T. Anti-inflammatory, tissue remodeling, immunomodulatory, and anticancer activities of oregano (Origanum vulgare) essential oil in a human skin disease model. Biochim Open. 2017;4:73-7. DOI: 10.1016/j.biopen.2017.02.005

Wei J, Huang Q, Bai F, Lin J, Nie J, Lu S, et al. Didymin induces apoptosis through mitochondrial dysfunction and up-regulation of RKIP in human hepatoma cells. Chem Biol Interact. 2017;261:118-26. DOI: 10.1016/j.cbi.2016.11.026

Forte C, Branciari R, Pacetti D, Miraglia D, Ranucci D, Acuti G, et al. Dietary oregano (Origanum vulgare L.) aqueous extract improves oxidative stability and consumer acceptance of meat enriched with CLA and n-3 PUFA in broilers. Poult Sci. 2018;97(5):1774-85. DOI: 10.3382/ps/pex452

Fokina A, Satarova T, Derkach K. The Effect of the mineral and carbohydrate composition of the nutrient medium on the efficiency of microclonal propagation of Origanum vulgare L. in vitro. Innov Biosyst Bioeng. 2019;3(3):176-84. DOI: 10.20535/ibb.2019.3.3.174795

Kalinin F, Sarnatskaya V, Polishchuk V. Methods of tissue culture in plant physiology and biochemistry. Kyiv: Naukova Dumka; 1980. 488 р.

Fokina A, Satarova T, Smetanin V, Kucenko N. Optimization of microclonal propagation in vitro of oregano (Origanum vulgare). Biosyst Divers. 2018;26(2):98-102. DOI: 10.15421/011815

Murashige T, Skoog F. A Revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant. 1962;15(3):473-97. DOI: 10.1111/j.1399-3054.1962.tb08052.x

Weaver K, Morales V, Dunn S, Godde K, Weaver P. An introduction to statistical analysis in research: with application in the biological and life sciences. Wiley; 2017. 594 р. DOI: 10.1002/9781119454205

Bondarev N, Reshetnyak O, Bondareva T, Il'in M, Nosov A. Impact of cultivation factors in vitro on the growth and the biosynthesis of steviol glycosides in Stevia rebaudiana cell cultures. Physiol Mol Biol Plants. 2019;25(4):1091-6. DOI: 10.1007/s12298-019-00680-6

Çakmak E, Uncuoğlu A, Aydın Y. Evaluation of in vitro genotoxic effects induced by in vitro anther culture conditions in sunflower. Plant Signal Behav. 2019;14(9):1633885. DOI: 10.1080/15592324.2019.1633885

Malik M, Mujib A, Gulzar B, Zafar N, Syeed R, Mamgain J, et al. Genome size analysis of field grown and somatic embryo regenerated plants in Allium sativum L. J Appl Genet. 2020;61(1):25-35. DOI: 10.1007/s13353-019-00536-5

Sinniah U, Mallappa K, Nakasha J, Kemat N. Induction, subculture cycle, and regeneration of callus in Safed musli (Chlorophytum borivilianum) using different types of phytohormones. Pharmacogn Mag. 2016;12(47):460-4. DOI: 10.4103/0973-1296.191457

Derkach K, Borysova V, Maletskyi V, Satarova T. The ability of maize Lancaster inbreds to callusogenesis in vitro under varying environmental conditions. Factors in Experimental Evolution of Organisms. 2018;22:228-34. DOI: 10.7124/FEEO.v22.953

Iconomou-Petrovich GN, Nianiou-Obeidat I. Micropropagation of Origanum vulgare subsp. hirtum (Mt. Taygetos). In: Tsekos I, Moustakas M, editors. Progress in Botanical Research. Dordrecht: Springer; 1998. P. 509-12. DOI: 10.1007/978-94-011-5274-7_116

Gamborg O, Murashige T, Thorpe T, Vasil I. Plant tissue culture media. In vitro. 1976;12:473-8. DOI: 10.1007/BF02796489

Goleniowski M, Flamarique C, Bima P. Micropropagation of oregano (Origanum vulgare×applii) from meristem tips. In vitro Cell Develop Biol Plant. 2003;39(2):125-8. DOI: 10.1079/IVP2002361

Habibi P, de Sa M, da Silva A, Makhzoum A, da Luz Costa J, Borghetti I, et al. Efficient genetic transformation and regeneration system from hairy root of Origanum vulgare. Physiol Mol Biol Plants. 2016;22(2):271-7. DOI: 10.1007/s12298-016-0354-2

El Beyrouthy M, Elian G, AbouJaoudeh C, Chalak L. In vitro propagation of Origanum syriacum and Origanum ehrenbergii. Acta Horticulturae. 2015;1083:169-72. DOI: 10.17660/ActaHortic.2015.1083.19



How to Cite

Fokina A, Denysiuk K, Satarova T. Origanum vulgare L. Cuttings Rhizogenesis in Microclonal Reproduction in Vitro. Innov Biosyst Bioeng [Internet]. 2020Apr.6 [cited 2023Jun.9];4(1):51-63. Available from: http://ibb.kpi.ua/article/view/192191