Investigation of the effects of carvacrol and nanocarvacrol on the expression of aflatoxin biosynthesis pathway genes in Aspergillus parasiticus using RT-qPCR

Mohamadnezhad , Sara; Nikaein , Donya; Khosravi , Alireza; Sharifzadeh , Aghil

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Volume 34, Issue 1 (3-2026)

Journal of Ilam University of Medical Sciences 2026, 34(1): 90-105

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Investigation of the effects of carvacrol and nanocarvacrol on the expression of aflatoxin biosynthesis pathway genes in Aspergillus parasiticus using RT-qPCR

Sara Mohamadnezhad ^*¹

, Donya Nikaein²

, Alireza Khosravi²

, Aghil Sharifzadeh²

1- Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran , mohamadnejad_s@yahoo.com
2- Dept of Microbiology and Immunology, Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

Abstract: (448 Views)

Introduction: Carvacrol is an important medicinal compound from the marjoram plant, whose antimicrobial properties on pathogenic and toxin-producing fungi have received little attention so far. Access to medicinal plants with nanocomposites provides the possibility of improving the properties and increasing the efficiency of medicinal plants in preventing the production of fungal toxins such as aflatoxin, given the limited availability of antifungal drugs and their resistance in the food and pharmaceutical industries. This study aimed to evaluate the effect of the antifungal properties of carvacrol and nanocarvacrol on the expression of aflatoxin biosynthesis genes, aflS, aflQ, aflD, and aflR, of Aspergillus parasiticus using RT-qPCR.
Materials & Methods: The standard strain of toxin-producing Aspergillus parasiticus (PTCC 5018) was used. SDA (Saboro Dextrose Agar) medium was used to evaluate the minimum inhibitory concentration (MIC). PDB (Potato Dextrose Agar) medium was used to perform HPLC and aflatoxin production. Dynamic light scattering (DLS) and Zeta analysis were used to examine the size and shape of particles with a scanning electron microscope to confirm the carvacrol nanoemulsion. To measure the expression of aflR, aflS, aflQ, and aflD genes, RNA extraction was performed using the guanidine isothiocyanate method. For cDNA synthesis, primers for afl genes and the 18S rRNA internal control gene were designed, and after cDNA synthesis, the RNA of the samples was examined using the RT-qPCR technique.
Results: In the presence of carvacrol and nanocarvacrol, the highest inhibition of fungal growth was observed at concentrations of 0.97 and 97 μg/μl, respectively. Using HPLC, a significant reduction in toxin production was determined by nanocarvacrol compared to carvacrol. The results of RT-qPCR also proved that nanocarvacrol inhibited aflatoxin-producing genes more than carvacrol at the molecular level.
Conclusion: The results showed that nanocarvacrol can inhibit fungal growth more than carvacrol and significantly reduced the expression of genes involved in aflatoxin biosynthesis in Aspergillus parasiticus.

Keywords: Nanocarvacrol, Aspergillus parasiticus, Gene expression, Aflatoxin biosynthesis, Zeta analysis

Full-Text [PDF 825 kb] (236 Downloads)

Editorial Note: Research | Subject: mycology
Received: 2025/04/28 | Accepted: 2026/01/20 | Published: 2026/03/25

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Mohamadnezhad S, Nikaein D, Khosravi A, Sharifzadeh A. Investigation of the effects of carvacrol and nanocarvacrol on the expression of aflatoxin biosynthesis pathway genes in Aspergillus parasiticus using RT-qPCR. J. Ilam Uni. Med. Sci. 2026; 34 (1) :90-105
URL: http://sjimu.medilam.ac.ir/article-1-8607-en.html

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