Cytochalasins in Diaporthe longicolla from Soybean ( Glycine max ): Insights from Untargeted Metabolomics and Implications for Animal Health

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Abstract
Species of Diaporthe cause soybean ( Glycine max L.) diseases in the United States, including stem canker, pod and stem blight, and seed decay. Among different species of Diaporthe , Diaporthe longicolla , the causal agent of pod and stem blight and seed decay, is associated with the production of cytochalasin compounds. In this study, untargeted metabolomic profiling was conducted on two isolates of D. longicolla (16-OP-SB-DIA-52 and 14-OP-PHO-SD-19) recovered from soybean plants affected by pod and stem blight to identify both known and previously uncharacterized metabolites. Axenic cultures were established in potato dextrose broth for metabolite characterization. Among the top 150 significantly upregulated metabolites compared with the potato dextrose broth control, nitro-phenoxy cytochalasin ( P = 0.009) and ortho-phenoxy cytochalasin ( P = 0.002) were putatively identified (Level 3 detection based on mass spectrometric parameters only). For 14-OP-PHO-SD-19, cytochalasin B ( P = 0.021) and nitro-phenoxy cytochalasin ( P = 0.002) were putatively detected (Level 3). In addition, metabolomic analysis putatively identified (Level 3) fumonisin B2 (16-OP-SB-DIA-52 [ P = 0.001]; 14-OP-PHO-SD-19 [ P = 0.009]) and zearalenone (16-OP-SB-DIA-52 [ P = 0.0002]; 14-OP-PHO-SD-19 [ P = 0.01]), which were significantly upregulated and among the top 400 metabolites in the two isolates. Interestingly, cytochalasin compounds, fumonisins, and zearalenone have been previously implicated in detrimental effects on livestock health (e.g., sheep, horses, and pigs). Although incorporating soybean meal into livestock diets underscores the potential relevance of D. longicolla -associated metabolites to feed safety, the present study was designed as a proof-of-concept metabolomics investigation. Consequently, any implications for animal health should be considered preliminary and require confirmation through metabolite quantification, feed analyses, and toxicity studies. Copyright © 2026 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .
Funding Information
  • United Soybean Board (25-209-S-B-1-A)
  • USDA National Institute of Food and Agriculture (2025-70006-45308)
  • USDA National Institute of Food and Agriculture (ND02250)

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