Influence of Ag nanoparticles on physiological and biochemical aspects of callus of Thymus species and Zataria multiflora Boiss.



Thymus species have found remarkable importance in food and medicine industries. The present study investigates the potential effect of Ag nanoparticle elicitors on proliferation of callus, and production of carvacrol and thymol in Zataria multiflora and three Thymus species. Firstly, callus was induced on Murashige and Skoog (MS) medium containing 2 mg l−1 of 2, 4-dichlorophenoxy acetic acid (2,4-D) and 1 mg l−1 of kinetin (Kin)). Secondly, the effects of two different concentrations of Ag nanoparticles (4 and 8 mg l-1) were studied on callus growth and its secondary metabolites production. Results elucidated that after elicitation by 8 mg l-1 ofAg NPs, significantly the highest callus growth rate (CGR) (0.02 mm day-1), callus fresh mass (CFM) (0.99 g), and carvacrol (0.68 mg l-1) and thymol (11.09 mg l-1) content was achieved. Comparing different Thymus species, notably the greatest carvacrol and thymol amount was obtained in .kotschyanus Boiss. & Hohen. and T. Daenesis Čelak. at 8 mg l-1 concentration ofAg NPs. Hence, it is evident that the stimulation by NPs is dose-dependent. This study has potential to be commercially applied for the enhancement of pharmaceutical compounds in different species of Thymus.


Ag nanoparticles; Thymus species; Zataria multiflora; callus; carvacrol thymol

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Afshar, B., & Golkar, P. (2016). Mucilage synthesis by in vitro cell culture in different species of Alyssum. BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology, 97(2), 79–86.

Ahmad, M. A., Javed, R., Adeel, M., Rizwan, M., Ao, Q., & Yang, Y. (2020). Engineered ZnO and CuO nanoparticles ameliorate morphological and biochemical response in tissue culture regenerants of candyleaf (Stevia rebaudiana). Molecules, 25(6), 1356.

Ajungla, L., Patil, P. P., Barmukh, R. B., & Nikam, T. D. (2009). Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metel L. Indian Journal of Biotechnology, 8(3), 317–322.

Alharby, H. F., Metwali, E. M. R., Fuller, M. P., & Aldhebiani, A. Y. (2016). Impact of application of zinc oxide nanoparticles on callus induction, plant regeneration, element content and antioxidant enzyme activity in tomato (Solanum lycopersicum mill.) under salt stress. Archives of Biological Sciences, 68(4), 723–735.

Ali, A., Mohammad, S., Khan, M. A., Raja, N. I., Arif, M., Kamil, A., & Mashwani, Z. ur R. (2019). Silver nanoparticles elicited in vitro callus cultures for accumulation of biomass and secondary metabolites in Caralluma tuberculata. Artificial Cells, Nanomedicine and Biotechnology, 47(1), 715–724.

Al-Jibouri, A. M. J., Abd, A. S., Majeed, D. M., & Ismail, E. N. (2012). Influence of abiotic elicitors on accumulation of thymol in callus cultures of Origanum vulgare L. Journal of Life Sciences, 6(10), 1094.

Asadollahei, M. V., Yousefifard, M., Tabatabaeian, J., Nekonam, M. S., & Mahdavi, S. M. E. (2022). Effect of elicitors on secondary metabolites biosynthesis in Zataria multiflora Boiss. Industrial Crops and Products, 181, 114789.

Barbasz, A., Kreczmer, B., & Oćwieja, M. (2016). Effects of exposure of callus cells of two wheat varieties to silver nanoparticles and silver salt (AgNO3). Acta Physiologiae Plantarum, 38(3), 1–11.

Castro, A. H. F., Braga, K. de Q., de Sousa, F. M., Coimbra, M. C., & Chagas, R. C. R. (2016). Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae). Revista Ciencia Agronomica, 47(1), 143–151.

Dykman, L. A., & Shchyogolev, S. Y. (2017). Interactions of plants with noble metal nanoparticles. Agricultural Biology, 52(1), 13.

Elumalai, E. K., Prasad, T. N. V. K. V., Hemachandran, J., Viviyan Therasa, S., Thirumalai, T., & David, E. (2010). Extracellular synthesis of silver nanoparticles using leaves of Euphorbia hirta and their antibacterial activities. Journal of Pharmaceutical Sciences and Research, 2(9), 549–554.

Ewais, E. ., Desouky, S., & Elshazly, E. . (2015). Evaluation of callus responses of Solanum nigrum L. exposed to biologically synthesized silver nanoparticles. Nanoscience and Nanotechnology, 5(3), 45–56.

Fazal, H., Abbasi, B. H., Ahmad, N., & Ali, M. (2016). Elicitation of medicinally important antioxidant secondary metabolites with silver and gold nanoparticles in callus cultures of Prunella vulgaris L. Applied Biochemistry and Biotechnology, 180(6), 1076–1092.

Fazal, H., Abbasi, B. H., Ahmad, N., Ali, M., Shujait Ali, S., Khan, A., & Wei, D. Q. (2019). Sustainable production of biomass and industrially important secondary metabolites in cell cultures of selfheal (Prunella vulgaris L.) elicited by silver and gold nanoparticles. Artificial Cells, Nanomedicine and Biotechnology, 47(1), 2553–2561.

Golkar, P., Bakhtiari, M.A. & Bazarganipour, M., (2021). The effects of nanographene oxide on the morpho-biochemical traits and antioxidant activity of Lepidium sativum L. under in vitro salinity stress. Scientia Horticulturae, 288, p.110301.

Goswami, L., Kim, K. H., Deep, A., Das, P., Bhattacharya, S. S., Kumar, S., & Adelodun, A. A. (2017). Engineered nano particles: Nature, behavior, and effect on the environment. Journal of Environmental Management, 196, 297–315.

Jasim, B., Thomas, R., Mathew, J., & Radhakrishnan, E. K. (2017). Plant growth and diosgenin enhancement effect of silver nanoparticles in Fenugreek (Trigonella foenum-graecum L.). Saudi Pharmaceutical Journal, 25(3), 443–447.

Javed, R., Ahmed, M., Haq, I. ul, Nisa, S., & Zia, M. (2017). PVP and PEG doped CuO nanoparticles are more biologically active: Antibacterial, antioxidant, antidiabetic and cytotoxic perspective. Materials Science and Engineering C, 79, 108–115.

Javed, R., Usman, M., Tabassum, S., & Zia, M. (2016). Effect of capping agents: Structural, optical and biological properties of ZnO nanoparticles. Applied Surface Science, 386, 319–326.

Javed, R., Yucesan, B., Zia, M., & Gurel, E. (2018). Elicitation of secondary metabolites in callus cltures of Stevia rebaudiana Bertoni grown under ZnO and CuO nanoparticles stress. Sugar Tech, 20(2), 194–201.

Kianersi, F., Pour-Aboughadareh, A., Majdi, M., & Poczai, P. (2021). Effect of methyl jasmonate on thymol, carvacrol, phytochemical accumulation, and expression of key genes involved in thymol/carvacrol biosynthetic pathway in some Iranian Thyme Species. International jJournal of Molecular Sciences, 22(20), 11124.

Kim, D. H., Gopal, J., & Sivanesan, I. (2017). Nanomaterials in plant tissue culture: the disclosed and undisclosed. RSC advances, 7(58), 36492-36505.

Kim, D., Jeong, S., & Moon, J. (2006). Synthesis of silver nanoparticles using the polyol process and the influence of precursor injection. Nanotechnology, 17(16), 4019–4024.

Kokina, I., Gerbreders, V., Sledevskis, E., & Bulanovs, A. (2013). Penetration of nanoparticles in flax (Linum usitatissimum L.) calli and regenerants. Journal of Biotechnology, 165(2), 127–132.

Lala, S. (2021). Nanoparticles as elicitors and harvesters of economically important secondary metabolites in higher plants: A review. IET Nanobiotechnology, 15(1), 28-57.

Mandeh, M., Omidi, M., & Rahaie, M. (2012). In Vitro influences of TiO2 nanoparticles on barley (Hordeum vulgare L.) tissue culture. Biological Trace Element Research, 150(1–3), 376–380.

Marslin, G., Sheeba, C. J., & Franklin, G. (2017). Nanoparticles alter secondary metabolism in plants via ROS burst. Frontiers in Plant Science, 8, 832.

Mathela, C. S., Singh, K. K., & Gupta, V. K. (2010). Synthesis and in vitro antibacterial activity of thymol and carvacrol derivatives. Acta Poloniae Pharmaceutica - Drug Research, 67(4), 375–380.

Miraj, S., & Kiani, S. (2016). Study of pharmacological effect of Ocimum basilicum: A review. Der Pharmacia Lettre, 8(9), 315–320.

Mosavat, N., Golkar, P., Yousefifard, M., & Javed, R. (2019). Modulation of callus growth and secondary metabolites in different Thymus species and Zataria multiflora micropropagated under ZnO nanoparticles stress. Biotechnology and Applied Biochemistry, 66(3), 316–322.

Murashige, T., & Skoog, F. (1962). A Revised Medium for Rapid Growth and Bio Agsays with Tohaoco Tissue Cultures. Physiologia Plantarum, 15(3), 474–497.

Ramakrishna, A., & Ravishankar, G. A. (2011). Influence of abiotic stress signals on secondary metabolites in plants. Plant Signaling and Behavior, 6(11), 1720–1731.

Rastogi, A., Zivcak, M., Sytar, O., Kalaji, H. M., He, X., Mbarki, S., & Brestic, M. (2017). Impact of metal and metal oxide nanoparticles on plant: A critical review. Frontiers in Chemistry, 5, 78.

Rechinger, K. (1982). Flora Iranica, Vol. 150. IC Hedge - Graz: Akademische Druck Und Verlagsanstalt, 150, 543–544.

Sadak, M. S. (2019). Impact of silver nanoparticles on plant growth, some biochemical aspects, and yield of fenugreek plant (Trigonella foenum-graecum). Bulletin of the National Research Centre, 43(1), 1-6.

Sajed, H., Sahebkar, A., & Iranshahi, M. (2013). Zataria multiflora Boiss. (Shirazi thyme) - An ancient condiment with modern pharmaceutical uses. Journal of Ethnopharmacology, 145(3), 686–698.

Sanzari, I., Leone, A., & Ambrosone, A. (2019). Nanotechnology in plant science: to make a long story short. Frontiers in Bioengineering and Biotechnology, 7, 120.

Shakya, P., Marslin, G., Siram, K., Beerhues, L., & Franklin, G. (2019). Elicitation as a tool to improve the profiles of high-value secondary metabolites and pharmacological properties of Hypericum perforatum. Journal of Pharmacy and Pharmacology, 71(1), 70–82.

Sharafi, E., Fotokian, M. H., & Loo, H. (2013). Improvement of hypericin and hyperforin production using zinc and iron nano-oxides as elicitors in cell suspension culture of John’swort (Hypericum perforatum L). Journal of Medicinal Plants and By-products, 2(2).

Sharma, P., Bhatt, D., Zaidi, M. G. H., Saradhi, P. P., Khanna, P. K., & Arora, S. (2012). Silver nanoparticle-mediated enhancement in growth and antioxidant status of Brassica juncea. Applied Biochemistry and Biotechnology, 167(8), 2225–2233.

Syu, Y. yu, Hung, J. H., Chen, J. C., & Chuang, H. wen. (2014). Impacts of size and shape of silver nanoparticles on Arabidopsis plant growth and gene expression. Plant Physiology and Biochemistry, 83, 57–64.

Zafar, H., Ali, A., Ali, J. S., Haq, I. U., & Zia, M. (2016). Effect of ZnO nanoparticles on Brassica nigra seedlings and stem explants: Growth dynamics and antioxidative response. Frontiers in Plant Science, 7, 535.

Zaka, M., Abbasi, B. H., Rahman, L.-U., Shah, A., & Zia, M. (2016). Synthesis and characterisation of metal nanoparticles and their effects on seed germination and seedling growth in commercially important Eruca sativa. IET Nanobiotechnology, 10(3), 1–7.

Zarshenas, M. M., & Krenn, L. (2015). A critical overview on Thymus daenensis Celak.: phytochemical and pharmacological investigations. Journal of Integrative Medicine, 13(2), 91–98.

Zhao, J., Davis, L., & Verpoorte, R. (2005). Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnology Advances, 23(4), 283–333.

Zuverza-Mena, N., Martínez-Fernández, D., Du, W., Hernandez-Viezcas, J. A., Bonilla-Bird, N., López-Moreno, M. L., Gardea-Torresdey, J. L. (2017). Exposure of engineered nanomaterials to plants: Insights into the physiological and biochemical responses-A review. Plant Physiology and Biochemistry, 110, 236–264.



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