Effects of Green-Synthesized ZnO Nanoparticles on Secondary Metabolite Accumulation in Alfalfa under Fusarium oxysporum Infection
Abstract
Root rot caused by several Fusarium species is a common and serious plant disease that threatens global agricultural products. To combat this type of disease, various methods are used in agriculture, such as pesticides, field sanitation, and the selection of resistant varieties. It is known that nanoparticles can elicit plant defense mechanisms. This study aimed to investigate the role of green-synthesized zinc oxide nanoparticles (ZnO NPs) in modulating the secondary metabolism of Medicago sativa (alfalfa) after Fusarium oxysporum infection. In our study, the total phenolic, flavonoid, and saponin contents of M. sativa plants treated with ZnO NPs were determined under F. oxysporum infection by using a spectrophotometer. According to results, while F. oxysporum inoculation increased total phenolic content (TPC) and total saponin content (TSC), the exogenous application of ZnO NPs significantly altered these responses during the infection. Notably, 1 ppm of ZnO NPs causes a dramatic increase in TSC, reaching 58 mg AE/g, suggesting a targeted stimulation of the isoprenoid pathway and triterpenoid saponin biosynthesis in M. sativa. In contrast, both 1 ppm and 2 ppm ZnO treatments result in a reduction in TSC compared to infected and non-infected controls. Similar to TSC, the total flavonoid content decreased dramatically in 1 ppm ZnO treatment, but no change was observed at 2 ppm ZnO concentration. These findings showed the potential of low-dose ZnO NPs as an effective strategy to enhance the innate chemical defense of alfalfa against F. oxysporum.
Keywords
green-synthesized, nanoparticle, ZnO, Medicago sativa, Fusarium oxysporum
Ethical Statement
References
- Akçay, M., Geyik, M. S., Yazicilar, B., Boke, F., Nadaroglu, H., Atıcı, O., & Bezirganoğlu, İ. (2025). Assessment of the effects of newly fabricated CaO, CuO, ZnO nanoparticles on callus formation maintenance of Alfalfa (Medicago sativa L.) under in vitro salt stress. Agronomy, 15(1), 180.
- Al-Darwesh, M. Y., Ibrahim, S. S., & Mohammed, M. A. (2024). A review on plant extract mediated green synthesis of zinc oxide nanoparticles and their biomedical applications. Results in Chemistry, 7, 101368.
- Arif, T., Bhosale, J. D., Kumar, N., Mandal, T. K., Bendre, R. S., Lavekar, G. S., & Dabur, R. (2009). Natural products–antifungal agents derived from plants. Journal of Asian Natural Products Research, 11(7), 621-638.
- Awuchi, C. G., Ondari, E. N., Ogbonna, C. U., Upadhyay, A. K., Baran, K., Okpala, C. O. R., ... & Guiné, R. P. (2021). Mycotoxins affecting animals, foods, humans, and plants: Types, occurrence, toxicities, action mechanisms, prevention, and detoxification strategies—A revisit. Foods, 10(6), 1279.
- Holghoomi, R., & Colagar, A. H. (2024). Applications of biocompatible nanoparticles in plant biotechnology for enhanced secondary metabolite biosynthesis. Inorganic Chemistry Communications, 167, 112753.
- Hussain, F., Hadi, F., & Akbar, F. (2019). Magnesium oxide nanoparticles and thidiazuron enhance lead phytoaccumulation and antioxidative response in Raphanus sativus L. Environmental Science and Pollution Research, 26(29), 30333-30347.
- Ibrahim, N. A., Rathore, D., Janiyani, K., Gupta, A., Sulieman, A. M. E., Tahir, H. E., ... & Surti, M. (2025). A comprehensive review on plant-derived bioactive saponins as promising antimicrobial agents: from bioavailability challenges, molecular mechanistic insights to therapeutic applications. Naunyn-Schmiedeberg's Archives of Pharmacology, 399(2), 1657-1687.
- Jarecka, A., Saniewska, A., Bialy, Z., & Jurzysta, M. (2008). The effect of Medicago arabica, M. hybrida, and M. sativa saponins on the growth and development of Fusarium oxysporum Schlecht f. sp. tulipae apt. Acta Agrobotanica, 61(2), 135-141.
- Karunarathna, S. C., Maharachchikumbura, S. S., Ariyawansa, H. A., Shenoy, B. D., & Jeewon, R. (2021). Emerging fungal plant pathogens. Frontiers in Cellular and Infection Microbiology, 11, 765549.
- Le, A. V., Parks, E. S., Nguyen, M. H., & Roach, P. D. (2018). Improving the vanillin-sulphuric acid method for quantifying total saponins. Technologies, 6(3), 84.