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Temporal wheat proteome remodeling by deoxynivalenol reveals novel detoxification signatures across cultivars

ABSTRACT: Fusarium head blight is a globally devastating fungal disease resulting in reduced grain yield and quality, along with contamination of grains with dangerous mycotoxins. Consumption of such mycotoxins by humans through processed food or livestock through feed has downstream implications of human and animal health. This interconnectivity across the environment, animal, and human health defines the One Health problem of threatened food safety and security. In this study, we explore remodeling of the wheat proteome upon exposure to a common mycotoxin, deoxynivalenol. We investigate cultivar specific responses to DON exposure in FHB-susceptible (Norwell) and -resistant (Sumai#3) cultivars across a continuum of exposure (i.e., 24 and 120 hours post inoculation), and upon low (i.e., 0.1 mg/mL) and high (1.0 mg/mL) levels of the mycotoxin. This complex experimental design enables us to tease apart the dynamic relationship between cultivar and DON tolerance. Specifically, we define specific proteins and broad categories of remodeling that are common (i.e., reduction in photosynthesis) and unique (i.e., glycosyltransferase) to the cultivars and align with anticipated protective mechanisms. Moreover, we applied an in vitro DON tolerance expression system and determined that induction of ubiquinol oxidase provides heightened protection for yeast growth relative to the negative control, as well as increased protection compared to a well-defined DON detoxifying protein. Our study suggests a new avenue for identification and characterization of novel DON detoxifying proteins as putative biomarkers for selected breeding strategies. Such strategies support the production of wheat varieties with increased tolerance to DON for improved global food safety and security.

INSTRUMENT(S):

ORGANISM(S): Triticum Aestivum (wheat)

TISSUE(S): Spikelet

SUBMITTER: Jennifer Geddes-McAlister

LAB HEAD: Jennifer Geddes-McAlister

PROVIDER: PXD059826 | Pride | 2025-06-08

REPOSITORIES: Pride

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			Action	DRS
	Data_guide.txt	Txt
	Nov_26_2021_RB_Plant_TMT_1.mzML	Mzml
	Nov_26_2021_RB_Plant_TMT_1.raw	Raw
	Nov_26_2021_RB_Plant_TMT_2.mzML	Mzml
	Nov_26_2021_RB_Plant_TMT_2.raw	Raw

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Project description:Mycotoxins are secondary metabolites produced by a variety of fungi that contaminate animal food and feeds and are capable of inducing a wide range of toxicities. Predictive in vitro models represent valuable substitutes for animal experiments to assess the toxicity of mycotoxins. The complexities of the interactions between epithelial and innate immune cells, vital for upholding barrier integrity and averting infections, remain inadequately understood. In the current study, a co-culture model of bovine epithelial cells (MAC-T) and macrophages (BoMac) was used to investigate the impact of exposure toFusariummycotoxins, namely deoxynivalenol (DON), zearalenone (ZEN), enniatin B (ENB), and beauvericin (BEA), on the inflammatory response elicited by the bacterial lipopolysaccharide (LPS) endotoxin. The MAC-T cells and BoMac were seeded on the apical side of a Transwell membrane and in the lower chamber, respectively, and mycotoxin exposure on the apical side of the membrane was carried out with the different mycotoxins (LC20; concentrations that elicited 20% cytotoxicity) for 48 h followed by an LPS immunity challenge for 24 h. The culture supernatants were collected from the basolateral compartment and these samples were submitted for cytokine/chemokine multiplex analysis. RNA-Seq analysis was performed using total RNA extracted from the MAC-T cells to acquire a more detailed insight into their cellular functions. The multiplex analysis indicated that IFN-γ, IL-1α, IL-8, and MCP-1 were significantly induced post-DON treatment when compared to control cells, and levels of IL-1α and IL-8 were enhanced significantly in all mycotoxin-treated groups post-LPS challenge. Analysis of the sequencing results showed that there were 341, 357, and 318 differentially expressed MAC-T cell genes that were up-regulated in the DON, ENB, and BEA groups, respectively. Gene ontology and pathway analysis revealed that these DEGs were significantly enriched in various biological processes and pathways related to inflammation, apoptosis signaling, and Wnt signaling. These results provide a comprehensive analysis of the co-culture cytokine/chemokine production and MAC-T cells’ gene expression profiles elicited byFusariummycotoxins, which further contributes to the understanding of early endotoxemia post-mycotoxin exposure.

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Temporal wheat proteome remodeling by deoxynivalenol reveals novel detoxification signatures across cultivars

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