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Abd Ellatif, S.; Elshafie, H.; , .; Hafez, E.; Ibrahim, A. Antifungal Activity, Gene Expression of Mentha Essential Oils. Encyclopedia. Available online: https://encyclopedia.pub/entry/22019 (accessed on 07 February 2026).
Abd Ellatif S, Elshafie H,  , Hafez E, Ibrahim A. Antifungal Activity, Gene Expression of Mentha Essential Oils. Encyclopedia. Available at: https://encyclopedia.pub/entry/22019. Accessed February 07, 2026.
Abd Ellatif, Sawsan, Hazem Elshafie,  , Elsayed Hafez, Amira Ibrahim. "Antifungal Activity, Gene Expression of Mentha Essential Oils" Encyclopedia, https://encyclopedia.pub/entry/22019 (accessed February 07, 2026).
Abd Ellatif, S., Elshafie, H., , ., Hafez, E., & Ibrahim, A. (2022, April 20). Antifungal Activity, Gene Expression of Mentha Essential Oils. In Encyclopedia. https://encyclopedia.pub/entry/22019
Abd Ellatif, Sawsan, et al. "Antifungal Activity, Gene Expression of Mentha Essential Oils." Encyclopedia. Web. 20 April, 2022.
Antifungal Activity, Gene Expression of Mentha Essential Oils
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Tomato (Lycopersicon esculentum Mill.) is important food in daily human diets. Root rot disease by Fusarium oxysporum caused huge losses in tomato quality and yield annually. The extensive use of synthetic and chemical fungicides has environmental risks and health problems. Recent studies have pointed out the use of medicinal plant essential oils (EOs) and extracts for controlling fungal diseases. In the current research, Mentha spicata and Mentha longifolia EOs were used in different concentrations to control F. oxysporum. Many active compounds are present in these two EOs such as: thymol, adapic acid, menthol and menthyl acetate. These compounds possess antifungal effect through malformation and degradation of the fungal cell wall. The relative expression levels of distinctly upregulated defense-related WRKY genes (WRKY1, WRKY4, WRKY33 and WRKY53) in seedling root were evaluated as a plant-specific transcription factor (TF) group in different response pathways of abiotic stress. Results showed significant expression levels of WRKY, WRKY53, WRKY33, WRKY1 and WRKY4 genes. An upregulation was observed in defense-related genes such as chitinase and defensin in roots by application EOs under pathogen condition. In conclusion, M. spicata and M. longifolia EOs can be used effectively to control this plant pathogen as sustainable and eco-friendly botanical fungicides

essential oils Fusarium root rot Mentha spicata Mentha longifolia GC–MS antioxidant enzymes , antifungal activity

1. Introduction

The tomato plant (Lycopersicon esculentum Mill.) is considered one of the third most important vegetable plants worldwide. Moreover, it is one of the most widespread vegetable crops grown across the globe. The tomato plant is highly sensitive to various biotic and abiotic stresses, which results in high economic losses [1][2]. The biotic stress which affects tomato plant growth and production is Fusarium oxysporum f. sp. lycopersici (Fol) [3]Fusarium oxysporum is a soil-borne pathogen that targets the plant by attacking the tomato roots, resulting in wilt disease [4]. Wilt disease frequency in tomato crops is very high in some countries where it reaches up to 25 ± 5% [5][6].
Additionally, in the presence of suitable conditions for the fungus, especially in developing countries, economic losses may increase up to 80% [7]. Consequently, the fungus gains its capability in tomato infection through secretion of mycotoxins [8], which have hazardous effects on animal and human health [9]. Mint essential oil (EO) has been reported to have a strong antimicrobial activity against several pathogenic microorganisms [10][11]. Many researchers have studied the biological activity of different EOs from Mentha against different pathogenic fungi, especially Fusarium species [11][12][13][14][15]. In particular, the two studied Mentha EOs have illustrated a strong antifungal activity against potato pathogens in addition to different soil-borne diseases in tomatoes [16][17][18].
The most active chemical compounds in EOs of Mentha species are piperitone oxide, pulegone, and 3-cyclopenten-1-one, 2-hydroxy-3-(3-methyl-2-butenyl) [19][20]. These compounds play an important role in defense against pests, pathogens, and fungi [21][22][23]. Sharma et al. [24] studied the effect of mint, clove, lemongrass, and eucalyptus EOs on wilt-causing fungus F. oxysporum. Plants’ EOs have a vital role in enhancing plant defense systems by increasing the production of phytochemicals as phenolic compounds and peroxidases enzymes which lead to strengthening of the cell wall and increasing lignification against phytopathogens [25].
When any pathogen infects plants, it is well known that they induce a plant’s defense system which works to resist both the pathogen attack and development of disease [26]. The plant defense system works once the plant is exposed to any stress; plant transcription factors belonging to multiple families play a critical role in stress mitigation or other adjustment mechanisms by modulating the gene expression patterns [27]. There is a large gene family, “WRKY”, which is considered the transcriptional factors distributed in all plant parts [28]. In addition, the WRKY genes were previously discovered in non-photosynthetic eukaryotes [29], and consequently have been identified and characterized in different plant species [30][31].
The main role of WRKY genes is defense; these genes work in the plant acquired resistance by using different pathways, including different enzymes [32]. Researchers have reported that they play a role in the defense mechanism of the Arabidopsis plant infected with necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola [33]. Several studies revealed that WRKY genes might bind with the promoter of phytoalexin deficient 3 and 1-aminocyclopropane-1-carboxylic acid synthase 2 when the plant is attacked by Botrytis cinerea [34].

2. Essential Oils Controlling Fusarium oxysporum

F. oxysporum f. sp. radicis-lycopersici is a wide-spread fungus in the plant rhizosphere which causes Fusarium crown and root rot (FCRR) disease and leads to losses of tomato production even in greenhouses and systems of soil production [35]. There are different management methods for root rot disease of tomato crop using chemical and biological controls [36]. Biological control for pathogenic fungi is the new management trend for reducing the harmful effects of chemicals (fungicides) [37][38]. There are four types of biocontrol management: microorganisms, semi-chemical products, plant-based natural products, or living microorganisms [39][40][41][42][43]. Furthermore, plant EOs are effective biocontrol agents against a variety of pathogenic fungi and bacteria [44][45][46][47].
The effect of Mentha spicata and M. longifolia on EOs on root rot disease of tomato infected with F. oxysporum both in vitro and in vivo. M. spicata and M. longifolia EOs had potentiality against Fusarium. The order of efficient EOs against Fusarium pathogen was: M. longifolia > M. spicata. The highest antifungal activity was observed in the case of all concentrations of M. longifolia EO in agreement with previous studies [48]. The capability of two Mentha EOs against fusarium is due to the ability of bioactive chemical molecules to penetrate the fungal cell wall and cytoplasmic membrane and destroy mitochondrial membranes [49]. Plant EOs contributed to loss of rigidity of the hyphal cell wall as well as damaging the cellular enzyme system, resulting in cell death [50][51]. Many other studies reported that the antifungal activity of M. spicata EO against F. oxysporum and Aspergillus niger depends on the chemical constituents: menthol, thymol, and piperitone, individually or in synergic effect [52][53]. Thymol compound activity presented in the malformation of the cellular membrane in addition to inhibition for ATPase activity [43][54]; regarding the effect of thymol and eugenol, they correlated to the ability of thymol compounds’ lysis of the external membrane of microorganisms which facilitated the entrance of eugenol to cytoplasm and interacted with protein [44][53]. Krishna Kishore et al. [55] demonstrated the antifungal activity of carvacrol, -terpineol, terpinen-4-ol, and linalool against Rhizoctonia solani, F. oxysporum, Penicillium digitatum, A. niger, Alternaria alternate, and A. flavus; these produce an effect against different microbial cells due to the ability of these compounds to penetrate the cell membrane, inactivate the enzyme pathway, and disturb their active transport [56][57].
The biological effect of the studied EOs on physiological parameters of tomato seedling is due to the presence of terpenes, alcohols, and phenolic compounds [55][58][59][60]. Moreover, the infected plants treated with M. longifolia EO showed the highest values of plant height (19.86 cm), shoot fresh weight (13.64 g), shoot dry weight (1.85 g), root fresh weight (1.41 g), and root dry weight (0.14 g). The main components in M. spicata, adipic acid 25.82% and piperitone 24.76%, are different than their percentage in M. spicata EO as reported by Bayan and Küsek [61]. Chemical constituents for M. longifolia EO were d-limonene, menthol, menthyl acetate, linalool, and eugenol, with percentages that differ from those reported from GC–MS analysis conducted by Desam et al. [62]. This difference in the percentage of single constituents for Mentha EOs may be due to differences in extraction methods or genetic diversity of these plants [63].
Menthol, menthyl acetate, linalool, and eugenol constituents alter cell permeability for Fusarium fungi and cause plasmolysis and cell death [64]. This study recorded collapsing of mycelium hyphae for F. oxysporum f. sp. lycopersici treated with EOs of M. spicata and M. longifolia with potential activity as biological control and therapeutic effect against root rot disease.
To analyze the role of chitinase, defensin, WRKY1, WRKY4, WRKY33, and WRKY53 transcripts in L. esculentum plant defense against F. oxysporum fungal pathogen, the researchers analyzed their expression after pathogen infection, and pathogen infection and application of 1.25% M. spicata and 1.0% M. longifolia EOs treatments. The expression results showed over-expression by 57.24 fold and changes in the level of WRKY33 transcription factor were recorded in infected plants treated with 1.0% M. longfolia, followed with 57.16 and 56.4 fold changes in pathogen, and 1.25% M. spicata treatments, respectively, while minimum expression of WRKY33 2.43 fold was observed with 1.25% M. spicata EO compared with control. The expression profile of WRKY35 TF revealed a significant upregulation of 39.23 and 38.12 fold changes at the pathogen-infected plant under 1.0% M. longfolia and pathogen treatments, respectively, and the minimum fold change of 3.52 was at 1.25% M. spicata EO compared with control. Data obtained in this study showed upregulation in the WRKY4 TF expression of 37.65 and 32.95 change folds at the pathogen-infected plant under 1.0% M. longfolia and pathogen treatments, respectively. In comparison, WRKY1 TF expression patterns were 26.35 and 25.17 change folds at the pathogen-infected plant under 1.0% M. longfolia and pathogen treatments, respectively, as compared with minimum expression level of 4.23 folds with 1.0% M. longfolia. The results were in agreement with previous studies which reported that WRKY3 and WRKY4 encode two structurally similar WRKY proteins, and their expression was responsive to stress conditions. Stress-induced expression of WRKY4 but not WRKY3 was further enhanced by pathogen infection. These results strongly suggest that WRKY4 regulates crosstalk between SA and JA/ET-mediated signaling pathways and, as a result, plays opposite roles in resistance to the two different types of microbial pathogens. Interestingly, WRKY proteins such as WRKY4, WRKY33, and redundant WRKY18, WRKY40, and WRKY60 play a positive role in plant resistance to necrotrophic pathogens.
The expression of defense-related genes showed over-expression under pathogen infection conditions and with the pathogen under 1.0% of M. longfolia and 1.25% of M. spicata. In contrast, chitinase gene was upregulated with 31.25, 29.6, and 27.83 fold changes at pathogen with 1.25% M. spicata, pathogenated plants, and then pathogenated with 1.0% M. longfolia treatments, respectively. In addition, defensin gene expressed as 18.65, 16.16, and 15.76 fold changes at pathogen with 1.0% M. longfolia, pathogen with 1.25% M. spicata, and pathogen treatments compared with minimum 2.44 fold expression was recorded at 1.25% M. spicata EO treatment.
These results collectively indicate that overexpression of chitinase, defensin, and WRKY transcripts play a positive role in inducing plant resistance against F. oxysporum and working toward reducing disease severity. Additionally, WRKY transcripts and PR3, PR12 genes were further enhanced by pathogen infection, and they are already considered as a marker for the plant–microbe interaction.
WRKY family members have diverse regulatory mechanisms; their protein can be effectively combined with W-box elements and bind to acting elements to activate or inhibit the transcription of downstream target genes through the cis-acting mechanism [65]. Thus, WRKY as a transcription factor plays an important role in plant defense in response to attacks by several pathogens. The response works by activating the expression of resistance genes directly or indirectly. It has been reported that WRKY DNA binding proteins bind to the promoter region of Arabidopsis natriuretic peptide receptor 1 (NPR1), which activated the plant defense system [66]. Moreover, WRKY33 activates the plant resistance system against necrotrophic fungi Alternaria brassicicola and Botrytis cinerea [29], and it can regulate the SAR system in the infected plants and also the PR genes [67][68]. Moreover, the high expression of such transcription factors could regulate the plant pathogen sensitivity to mutants of AtWRKY4, AtWRKY3, and AtWRKY3 WRKY4, increasing the plant susceptibility toward the fungus B. cinerea. In contrast, the high expression of the non-mutated AtWRKY4 enhanced the plant’s resistance toward the Pseudomonas syringae [69]. Many plant WRKY genes are induced by biotrophic and necrotrophic pathogens, including fungi and viruses, through the induction of SA-dependent SAR and PR genes [70][71].

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