Influence of plant growth regulators and salicylic acid on the production of some secondary metabolites in callus and cell suspension culture of Satureja sahendica Bornm.



The impact of combinations of plant growth regulators (PGRs) on callus culture of Satureja sahendica Bornm. was investigated. In nodal explants, the response of secondary metabolite production to different concentrations of PGRs was analyzed regarding the presence and absence of polyvinylpyrrolidone (PVP). The explants were cultured on MS media in presence of auxins (2,4-dichlorophenoxyacetic acid and naphthylacetic acid) and cytokinins (thidiazuron and kinetin); which were used in equal concentrations of 0.5, 1, and 2 mg l-1. The treatment of 2 mg l-1 2,4-D + 2 mg l-1 Kin (MD3) led to the highest production of total phenolics (4.303 ± 0.449 mg GAE g-1) and flavonoids (24.903 ± 7.016 mg QE g-1). Moreover, the effect of salicylic acid (SA) on the production of secondary metabolites in cell suspension culture of Satureja sahendica was evaluated. The cell suspension culture was established by culturing the nodal-derived friable callus in the liquid medium containing different concentrations of SA (0, 100, 150, 200 µM). An inverse relationship exists between the fresh mass and secondary metabolites contents. In addition, there was a significant difference among concentrations of SA in the production of total phenolics and flavonoid compounds. SA enhances secondary metabolites production and decreases cell fresh mass.


Satureja sahendica; callus induction; cell suspension culture; secondary metabolites; growth regulators; salicylic acid

Full Text:



Ali, M., Abbasi, B. H., Ul-haq, I. (2013). Production of commercially important secondary metabolites and antioxidant activity in cell suspension cultures of Artemisia absinthium L. Industrial Crops and Products, 49, 400-406.

Al-juboory, K. H., Skirvin, R.M., William, D.J. (1998). Callus induction and adventitious shoot regeneration of gardenia (Gardenia jasminoides Ellis) leaf explants. Scientia Horticulturae, 72(3-4), 171-178.

Al-Sane, K. O., Shibli, R.A., Freihat, N.M., Hammouri, M.K. (2005). Cell suspension culture and secondary metabolites production in African violet (Saintpaulia ionantha Wendl.). Jordan Journal of Agricultural Sciences, 1(1), 84-92.

Alvarez, P.S., Spollansky, T. C., Giulietti, A.M. (2000). The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmansia candida. Enzyme and Microbial Technology, 26(2-4), 254-258.

Antonigni, F., Zheng, S., Pagnucco, C., Baraldi, R., Poli, F., Biondi, S. (2007). Induction of flavonoid production by UV-B radiation in Passiflora quagrangularis callus cultures. Fitoterapia, 78(5), 345-352.

Chang, C. C., Yang, M.Y., Wen. H. M., Chern, J. C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis, 10(3), 178-182.

Dörnenburg, H., Knorr, D. (1995). Strategies for the improvement of secondary metabolite production in plant cell cultures. Enzyme and Microbial Technology, 17(8), 674-684.

Esmaeilzadeh Bahabadi, S., Rezaei, A. (2014). Increased trigonelline production by salicylic acid in fenugreek (Trigonella foenum-graecum L.) cell culture. (in Persian). Journal of Cell and Tissue, 5, 165-172.

Fu, C. X., Zhao, D.X., Huang, Y., Ma, F. S. (2005). Cellular aggregate size as the critical factor for flavonoid production by suspension cultures of Saussurea medusa. Biotechnology Letters, 27(2), 91-95.

Ghahreman, A. (1988). Flora of Iran (in Persian). Research Institute of Forest and Rangeland, Tehran.

Ghahreman, A. (1993). Plant Systematics: Cormophytes of Iran (in Persian). Iran University Press, Tehran.

Ghotbabadi, F. S., Alizadeh, A., Zadehbagheri, M., Kamelmanesh, M. M., Shaabani, M. (2012). Phytochemical composition of the essential oil, total phenolic content, antioxidant and antimicrobial activity in Iranian Satureja sahendica Bornm. at different ontogenesis conditions. Journal of Medicinal Plants Research, 6(19), 3525-3534.

Güllüce, M., Sokmen, M., Daferera, D., Ağar, G., Ӧzkan, H., Kartal, N., Polissiou, M., Sökmen, A., Sahin, F. (2003). In vitro antibacterial, antifungal, and antioxidant activities of the essential oil and methanol extracts of herbal parts and callus cultures of Satureja hortensis L. Journal of Agricultural and Food Chemistry, 51(14), 3958-3965.

Hadian, J., Akramian, M., Heydarian, H., Mumivan, H., Asghari, B. (2012). Composition and in vitro antibacterial activity of essential oils from four Satureja species growing in Iran. Natural Product Research, 26(2), 98-108.

Hadjmohammadi, M. R., Soltani, M., Sharifi, V. (2012). Use of hollow fiber liquid phase microextraction and HPLC for extraction and determination of apigenin in human urine after consumption of Satureja sahendica Bornm. Journal of Chromatography B, 900, 85-88.

Hakkim, F. L., Gowri-Shankar, C., Girija, S. (2007). Chemical composition and antioxidant property of holy basil (Ocimum sanctum L.) leaves stems, and inflorescence and their in vitro callus cultures. Journal of Agricultural and Food Chemistry, 55(22), 9109-9117.

Hakkim, F. L., Kalyani, S., Essa, M., Girija, S. Song, H. (2011). Production of rosmarinic acid in Ocimum sanctum (L.) cell suspension cultures by the influence of growth regulators. International Journal of Biological and Medical Research, 2(4), 1158-1161.

Han, K.L., Lee, Y., Song, J. H., Hwang, Y. S., Lee, W. S., Kim, M. W., Kim, S. H. (2012). Enhanced production and secretion of rutin and GABA in immobilized cells of mulberry tree (Morus bombycis K.). Plant Cell, Tissue and Organ Culture, 108(3), 513-20.

Harfouche, A. L., Rugini, E., Mencarelli, F., Botondi, R., Muleo, R. (2008). Salicylic acid induces H2O2 production and endochitinase gene expression but not ethylene biosynthesis in Castanea sativa in vitro model system. Journal of Plant Physiology, 165(7), 734-744.

Hassanpouraghdam, M. B., Safi-Shalamzari, M., Azami, M. A., Mohajjel-Shoja, A. (2009). γ-terpinene and carvacrol rich volatile oil of Satureja sahendica Bornm. from Maragheh district in Northwest Iran. Chemija, 20(3), 186-189.

James, J. T., Meyer, R., Dubery, I. A. (2008). Characterization of two phenotypes of Centella asiatica in Southern Africa through the composition of four triterpenoids in callus, cell suspensions and leaves. Plant Cell, Tissue and Organ Culture, 94(1), 91-99.

Jamzad, Z. (1996). Satureja rechingeri (Labiatae): A new stream. Species from Iran. (pp. 75-7). Annalen des Naturhistorischen Museums in Wien.

Jeong, G. T., Woo, J. C., Park, D. H. (2007). Effect of plant growth regulators on growth and biosynthesis of phenolic compounds in genetically transformed hairy roots of Panax ginseng C. A. Meyer. Biotechnology and Bioprocess Engineering, 12(2), 86-9.

Johnson, M., Wesely, E. G., Kavitha, M. S., Uma, V. (2011). Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L. Asian Pacific Journal of Tropical Medicine, 4(3), 196-200.

Khosroushahi, A. Y., Valizadeh, M., Gasempour, A., Khosroushahi, M., Naghdibadi, H., Dadpour, M. R., Omidi, Y. (2006). Improved taxol production by combination of inducing factor in suspension culture of Taxus baccata. Cell Biology International, 30(3), 262-269.

Kim, N. S., Jung, D. H., Jung, C. R., Jeon, K. S., Park, H. W., & Park, S. U. (2020). Improvement of phenylpropanoid production with elicitor treatments in Pimpinella brachycarpa Nakai. Horticulturae, 6(4), 108.

Kovacik, J., Backor, M., Strnad, M., Repcak, M. (2009). Salicylic acid-induced changes to growth and phenolic metabolism in Matricaria chamomilla plants. Plant Cell Reports, 28(1), 135-143.

Leyser, O., Day, S. (2003). Mechanism in plant development, Oxford, UK: Blackwell Science Ltd, ISBN: 978-0-86542-742-6.

Luczkeiwicz, M., Glod, D. (2003). Callus cultures of Genista plants- in vitro material producing high amounts of isoflavones of phytoestrogenic activity. Plant Science, 165(5), 1101-1108.

Matkowski, A. (2008). Plant in vitro culture for the production of antioxidants — A review. Biotechnology Advances, 26(6), 548-560.

Mendhulkar, V. D., Moinuddin, M., Vakil, A. (2013). Elicitation of flavonoids by salicylic acid and Penicillium expansum in Andrographis paniculata (Burm. f.) Nees. cell culture. Research in Biotechnology, 4(2), 1-9.

Moghadam, Y. A., Piri, K. H., Bahramnejad, B., & Habibi, P. (2013). Methyl jasmonate and salicylic acid effects on the dopamine production in hairy cultures of Portulaca oleracea (purslan). Bulletin of Environment, Pharmacology and Life Sciences, 2(6), 89-94.

Mozaffarian, V. (1993). A dictionary of Iranian plant names (pp. 482-483) Farhang Moaser Publication, Tehran, Iran.

Mozaffarian, V. (1996). A dictionary of Iranian plant names. Farhang Moaser Publication, Tehran, Iran.

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

Murthy, B. N. S., Murch, S. J. Saxena, P. K. (1998). Thidiazuron: A potent regulator of in vitro plant morphogenesis. In Vitro Cellular & Developmental Biology-Plant, 34(4), 267-275.

Namadeo, A. G. (2007). Plant cell elicitation for production of secondary metabolites— A review. Pharmacognosy Reviews, 1(1), 154-160.

Neumann, K. H., Kumar, A., Imani, J. (2009). Plant cell and tissue culture. A tool in biotechnology (pp. 333). Springer- Verlag Berlin Heidelberg.

Ogita, S., Sasamoto, H., Yeung, E. C., & Thorpe, T. A. (2001). The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica. In Vitro Cellular & Developmental Biology-Plant, 37(2), 268-273.

Pourebad, N., Motafakkerazad, R., Kosari-Nasab, M., Farsad-Akhtar, N., Movafeghi, A. (2015). The influence of TDZ concentrations on in vitro growth and production of secondary metabolites by the shoot and callus culture of Lallemantia iberica. Plant Cell, Tissue and Organ Culture, 122(2), 331-339.

Rajjou, L., Belghazi, M., Huguet. R., Robin, C., Moreau, A., Job, C. (2006). Proteomic investigation of the effect of salicylic acid on Arabidopsis seed germination and establishment of early defense mechanisms. Plant Physiology, 141(3), 910-23.

Rani, D. N., Nair, G. M. (2006). Effects of plant growth regulators on high frequency shoot multiplication and callus regeneration of an important Indian medicinal plant, Nirgundi (Vitix negundo L.). In Vitro Cellular and Developmental Biology-Plant, 42(1), 69-73.

Ravishandera, G. A., Bhyalakshmi, N., Ramachandra Rao, S. (1999). Production of food additive. Biotechnology: Secondary metabolites (pp. 89-110). New Dehli: Oxford IBH.

Rechinger, K. H. (1982). Satureja in Flora Iranica (pp. 495-504). Akademische Druck-u. Verlagsanstalt, Graz.

Ruiz-García, Y., & Gómez-Plaza, E. (2013). Elicitors: a tool for improving fruit phenolic content. Agriculture, 3(1), 33-52.

Sadeghian, F., Hadian, J., Hadavi, M., Mohamadi, A., Ghorbanpour, M., Ghafarzadegan, R. (2013). Effects of exogenous salicylic acid application on growth, metabolic activities and essential oil composition of Satureja khuzistanica Jamzad. Journal of Medicinal Plants, 3(47), 70-82.

Saeidnia, S., Nourbakhsh, M. S., Gohari, A. R., Davood, A. (2011). Isolation and identification of the main compounds of Satureja sahendica Bornm. Australian Journal of Basic and Applied Sciences, 5(6), 1450-1453.

Samadi, S., Ghasemnezhad, A., Alizadeh, M. (2014). Investigation on phenylalanine ammonia-lyase activity of artichoke (Cynara scolymus L.) affected by methyl jasmonate and salicylic acid in in-vitro conditions (in Persian). Journal of Plant Production Research, 21, 135-148.

Saxena, P. K., & Gill, R. (1986). Removal of browning and growth enhancement by polyvinylpolypyrrolidone in protoplast cultures of Cyamopsis tetragonoloba L. Biologia Plantarum, 28(4), 313-315.

Saxena, S., & Dhawan, V. (1999). Regeneration and large-scale propagation of bamboo (Dendrocalamus strictus Nees) through somatic embryogenesis. Plant Cell Reports, 18(5), 438-443.

Sefidkon, F., Jamzad, Z. Mirza, M. (2004). Chemical variation in the essential oil of Satureja sahendica from Iran. Food Chemistry, 88(3), 325-328.

Shilpashree, H., Rai, R. (2009). Effects of growth regulators on in vitro plant regeneration and flavonoid production in Hypericum mysorense. International Journal of Integrative Biology, 8(1), 43-49.

Singleton, V. L., Orthofer, R., Lamuela-Raventos, R. M. (1999). Analysis of total phenols and other oxidation substrates and oxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology, 299, 152-178.

Sivanandhan, G., Kapil Dev, G., Jeyaraj, M., Rajesh, M., Muthuselvam, M., Selvaraj, N., Manickavasagam, M., Ganapathi, A. (2013). A promising approach on biomass accumulation and withanolides production in cell suspension culture of Withania somnifera (L.) Dunal. Protoplasma, 250(4), 885-898.

Sun, Z., Hou, S., Yang, W., & Han, Y. (2012). Exogenous application of salicylic acid enhanced the rutin accumulation and influenced the expression patterns of rutin biosynthesis related genes in Fagopyrum tartaricum Gaertn leaves. Plant Growth Regulation, 68(1), 9-15.

Tarigholizadeh, S., Motafakkerazad, R., Kosari-Nasab, M., Movafeghi, A. (2015). Callus medium improvement for Satureja sahendica as an important medicinal plant. International Journal of Integrative Biology, 16(1), 13-18.

Taveira, M., Ferreres, F., Pereira, M. D., Sousa, C., Andrade, B. P., Martins, A., Pereira, A. J., Valentao, P. (2010). In vitro culture of Brassica oleracea L.var. costata DC: potential plant bioreactor for antioxidant phenolic compounds. Journal of Agricultural and Food Chemistry, 57(4), 1247-1252.

Tepe, B., Sokmen, A. (2007). Production and optimization of rosmarinic acid by Satureja hortensis L. callus cultures. Natural Product Research, 21(13), 1133-1144.

Thiruvengadam, M., Chung, I. M. (2015). Phenolic compound production and biological activities from in vitro regenerated plants of gherkin (Cucumis anguria L.). Electronic Journal of Biotechnology, 18(4), 295-301.

Tomlin, C. D. S. (2006). The Pesticide Manual: A World Compendium. 14th ed. British Crop Protection Council: Surrey, UK.

United States Environmental Protection Agency (US EPA). Reregistration eligibility decision for 2,4-D. EPA 738-R-05-002; June 2005b.

Vlot, A. C., Klessig, D. F., Park, S. W. (2008). Systemic acquired resistance: the elusive signal(s). Current Opinion in Plant Biology, 11(4), 436-442.

Walla Abdelazeez, M. A., Khusnetdinova Landysh, Z., Timofeeva Olga, A. (2017). Effect of different plant growth regulators on the in vitro induction and maintenance of callus from different explants of Hyoscyamus muticus L. Journal of Applied Environmental and Biological Sciences, 7(3), 27-35.

Wang, K., Jin, P., Cao, S., Shang, H., Yang, Z., Zheng, Y. (2009). Methyl jasmonate reduces decay and enhances antioxidant capacity in Chinese bayberries. Journal of Agricultural and Food Chemistry, 57(13), 5809–5850.

Wang, Y., Yuan, Y., Wn, J. (2004). Induction studies of methyl jasmonate and salicylic acid on taxane production in suspension cultures of Taxus chinensis var. mairei. Biochemical Engineering Journal, 19(3), 259-265.

War, A. R., Paulraj, M. G., War, M. Y., Ignacimuthu S. (2011). Jasmonic acid-mediated induced resistance in groundnut (Arachis hypogaea L.) against Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae). Journal of Plant Growth Regulation, 30(4), 512-523.

Wen, P. F., Chen, J. Y., Kong, W. F., Pan, Q. H., Wan, S. B., Huang, W. D. (2004). Salicylic acid induced the expression of phenylalanine ammonia-lyase gene in grape berry. Plant Science, 169(5), 928-934.

World Health Organization (WHO). Environmental Health Criteria 84, Environmental aspects - 2,4-dichlorophenoxyacetic acid (2,4-D); International Programs on Chemical Safety. World Health Organization: Geneva, Switzerland; 1989.

Zhong, J. J., Bai, Y., Wang, S. J. (1996). Effects of plant growth regulators on cell growth and ginsenoside saponin production by suspension cultures of Panax quinquefolium. Journal of Biotechnology, 45(3), 227-234.



  • There are currently no refbacks.

Copyright (c) 2021 Sarieh Tarigholizadeh, Rouhollah Motafakkerazad, Morteza Kosari-Nasab, Ali Movafeghi, Sakineh Mohammadi, Mohsen Sabzi Nojadeh, Amir-Hossein Talebpour

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.


Acta agriculturae Slovenica is an Open Access journal published under the terms of the Creative Commons CC BY License.


eISSN 1854-1941