Alternaria alternata is a critical phytopathogen that causes foodborne spoilage and produces a polyketide mycotoxin, alternariol (AOH), and its derivative, alternariol monomethyl ether (AME). In this study, the inhibitory effects of the essential oil citral on the fungal growth and mycotoxin production of A. alternata were evaluated. Our findings indicated that 0.25 μL/mL (222.5 μg/mL) of citral completely suppressed mycelial growth as the minimum inhibitory concentration (MIC). Moreover, the 1/2MIC of citral could inhibit more than 97% of the mycotoxin amount. Transcriptomic profiling was performed by comparative RNA-Seq analysis of A. alternata with or without citral treatment. Out of a total of 1334 differentially expressed genes (DEGs), 621 up-regulated and 713 down-regulated genes were identified under citral stress conditions. Numerous DEGs for cell survival, involved in ribosome and nucleolus biogenesis, RNA processing and metabolic processes, and protein processing, were highly expressed in response to citral. However, a number of DEGs responsible for the metabolism of several carbohydrates and amino acids, sulfate and glutathione metabolism, the metabolism of xenobiotics and transporter activity were significantly more likely to be down-regulated. Citral induced the disturbance of cell integrity through the disorder of gene expression, which was further confirmed by the fact that exposure to citral caused irreversibly deleterious disruption of fungal spores and the inhibition of ergosterol biosynthesis. Citral perturbed the balance of oxidative stress, which was likewise verified by a reduction of total antioxidative capacity. In addition, citral was able to modulate the down-regulation of mycotoxin biosynthetic genes, including pksI and omtI. The results provide new insights for exploring inhibitory mechanisms and indicate citral as a potential antifungal and antimytoxigenic alternative for cereal storage.

Alternaria alternata is an important phytopathogen causing fruit black rot and also producing a variety of mycotoxins, such as alternariol (AOH) and alternariol monomethyl ether (AME) as two main contaminants. This could lead to economic losses of agricultural products as well as human health risks. In this study, magnolol extracted from the traditional Chinese herb, Mangnolia officinalis, exhibited an obvious antifungal property and could completely suppress the mycelial growth at 100 μM. Morphological differences of A. alternata were observed to be significantly shrunk and wrinkled after the exposure to magnolol. Furthermore, AOH and AME were no longer produced in response to 50 μM of magnolol. To uncover the antifungal and antimycotoxigenic mechanisms, the transcriptomic profiles of A. alternata—treated with or without magnolol—were evaluated. The clustered genes responsible for AOH and AME biosynthesis were obviously less transcribed under magnolol stress and this was further confirmed by qRT-PCR. The global regulators of carbon and nitrogen utilization, such as CreA and NmrA, were significantly down-regulated and this possibly caused the reduction in mycotoxins. In addition, fatty acid β-oxidation was regarded to contribute to polyketide mycotoxin production for the supply of precursor acetyl-CoA while the expression of these related genes was inhibited. The response to magnolol led to the marked alteration of oxidative stress and the down-expression of the mitogen-activated protein kinase (MAPK) signaling pathway from the transcriptome data and the determination of peroxidase (POD), superoxide dismutase (SOD) and glutathione (GSH) assays. This above might be the very reason for the growth supression and mycotoxin production of A. alternata by magnolol. This study provides new insights into its potential as an important active ingredient for the control of A. alternata and its mycotoxins in fruits and their products.

One-step solid-phase extraction (SPE) using a multiwalled carbon nanotube (MWCNT) for simultaneous analysis of 21 mycotoxins, including nine trichothecenes, zearalenone (ZEN) and its derivatives, four aflatoxins, and two ochratoxins, in corn and wheat was developed. Several key parameters affecting the performance of the one-step SPE procedure—types of MWCNT, combinations with five sorbents (octadecylsilyl (C₁₈), hydrophilic–lipophilic balance (HLB), mixed-mode cationic exchange (MCX), silica gel, and amino-propyl (NH₂)), and filling amounts of the MWCNTs—were thoroughly investigated. The combination of 20 mg carboxylic MWCNT and 200 mg C₁₈ was proven to be the most effective, allowing the quantification of all analyzed mycotoxins in corn and wheat. Under the optimized cleanup procedure prior to ultraperformance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) analysis, the method was validated by analyzing samples spiked at the limit of quantification (LOQ), two-times LOQ, and 10-times LOQ. Satisfactory linearity (r² ≥ 0.9910), high sensitivity (LOQ in different ranges of 0.5–25 μg L⁻¹), good recovery (75.6–110.3%), and acceptable precision (relative standard deviation (RSD), 0.3–10.7%) were obtained. The applicability of the method was further confirmed using raw samples of corn and wheat. In conclusion, the established method was rapid, simple and reliable for simultaneous analysis of 21 mycotoxins in corn and wheat.

Fumonisins are polyketide-derived mycotoxins posing significant health threats to humans and animals. Among these, fumonisin B₁ (FB₁) is the most prevalent mycotoxin, primarily produced by Fusarium verticillioides, especially in maize and its derived products. Tangeretin, a polymethoxyflavonoid, has been identified as having potential medicinal properties, particularly as an antioxidant. To evaluate the antifungal and anti-mycotoxigenic properties of tangeretin and to elucidate the mechanisms underlying its inhibitory effects, assessments of fungal growth, FB₁ production, conidial germination, and cellulase activity, antioxidant capacity and enzyme activities, transcriptomic analysis and gene deletion experiments were conducted. Consequently, tangeretin significantly curtailed fungal growth and FB₁ production and provided protection against pathogenic infection on corn. It affected genes associated with fungal growth, conidial development, and antioxidant response. Furthermore, tangeretin interfered with the supply of biosynthetic substrate necessary for fumonisin production, particularly impacting pathways involved in alanine metabolism, pyruvate metabolism, fatty acid degradation, and sphingolipid metabolism. Notably, tangeretin downregulated four biosynthetic genes (Fum2, Fum3, Fum10 and Fum11) that are involved in the final steps of fumonisin formation. It likely disrupted the MAPK signaling pathway and targeted a putative NmrA- and HSCARG-like protein Fv_Tan1, which was identified as having positive effects on fungal growth and mycotoxin biosynthesis. This study presents a promising approach for controlling fumonisin contamination in agricultural settings.

Ochratoxin A (OTA) is a major mycotoxin contaminant in grapes and their products, and Aspergillus carbonarius is its main producer. Controlling the growth of A. carbonarius is therefore critical for mitigating OTA contamination. Plant-derived perillaldehyde, with good antifungal activity and safety, has garnered growing attention. However, current understanding of how perillaldehyde affects A. carbonarius growth and OTA production remains poorly characterized. In this study, we systematically investigated the antifungal and antimycotoxigenic effects of perillaldehyde against A. carbonarius and explored the underlying mechanisms. The results showed that perillaldehyde could alter the mycelial morphology and damage the cell integrity of A. carbonarius. Additionally, perillaldehyde could diminish the total antioxidant capacity and impair the energy metabolism of A. carbonarius. Transcriptome analysis showed that the expressions of all the known conserved OTA biosynthetic genes and two OTA transport-related genes were significantly down-regulated, indicating that perillaldehyde could directly inhibit their expression. In conclusion, perillaldehyde can significantly inhibit OTA production by directly disrupting OTA biosynthesis and transport and inhibiting the growth of A. carbonarius. Thus, perillaldehyde has the potential to be used as a natural fungicide or alternative food preservative in grapes and their products, owing to its strong antifungal and antimycotoxigenic effects on A. carbonarius.