DNA microarray to detect and identify trichothecene- and moniliformin-producing Fusarium species

Area of Science:

• Mycology
• Molecular Biology
• Food Safety

Background:

• Fusarium species are significant plant pathogens and producers of harmful mycotoxins like trichothecenes and moniliformin.
• Accurate and rapid detection of these fungi is crucial for agricultural and food safety applications.

Purpose of the Study:

• To develop a DNA microarray for the easy and fast detection and identification of trichothecene- and moniliformin-producing Fusarium species.
• To create a tool for differentiating between Fusarium species groups based on their mycotoxigenic potential.

Main Methods:

• Design of capture probes based on phylogenetic analysis of translation elongation factor-1 alpha (TEF-1alpha) gene sequences.
• A consensus PCR amplification of a TEF-1alpha fragment followed by hybridization to the developed Fusarium chip.
• Visualization of results using a colorimetric Silverquant detection method.

Main Results:

• Development of a DNA microarray capable of detecting and identifying 14 Fusarium species, with differentiation of four species groups.
• Validation against naturally infected cereal samples, demonstrating high sensitivity with a limit of detection below 16 genomic DNA copies.
• Successful differentiation of species based on their mycotoxigenic abilities.

Conclusions:

• The developed Fusarium chip is a highly sensitive and rapid tool for Fusarium species detection and identification.
• The method shows potential for (semi-)quantification of Fusarium DNA.
• The Fusarium chip is a valuable tool for screening cereal samples in the food and feed production chain, aiding in the detection of known and novel Fusarium species.