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High-Resolution Melting assays development for discrimination of fungal pathogens causing Grapevine Trunk Diseases

Summary

Grapevine Trunk Diseases cause major losses in vineyards worldwide but are difficult to diagnose early because symptoms may not appear for years. This study developed fast and affordable DNA-based tests called HRM assays that can identify ten different disease-causing fungi in grapevines. These tests work by analyzing DNA melting patterns and can even detect infections before symptoms appear, helping vineyard managers make better decisions about plant treatment and removal.

Background

Grapevine Trunk Diseases (GTDs) are a complex set of fungal diseases affecting wood tissues of grapevine, causing significant economic losses in the wine industry. Current diagnostic methods based on symptomatology are unreliable due to irregular symptom onset and quiescent infections that can remain dormant for years. Molecular methods are needed for early and accurate identification of GTD-causing pathogens.

Objective

To develop High-Resolution Melting (HRM) assays based on the beta-tubulin gene (Tub2) that can detect and identify the most common fungal species responsible for four major Grapevine Trunk Diseases. The assays aim to provide a fast, cost-effective tool for disease detection and management in vineyards.

Results

The three HRM assays successfully distinguished ten common GTD-causing fungal species based on melting curve profiles against Neofusicoccum parvum reference. Multi-assay analysis allowed clear identification of species causing Esca, Phomopsis dieback, and Eutypa dieback, and enabled both identification and relative quantification of pathogens in binary mixtures. Closely related species could be differentiated through combined analysis of curve profiles across all three assays.

Conclusion

A set of HRM-based assays was successfully developed for rapid identification of GTD-causing fungi, providing a quick, cost-effective diagnostic tool for vineyard management. This methodology enables early detection of quiescent infections and assessment of fungal load, allowing producers to implement timely management practices and reduce viticulture production costs.
  • Published in:PLoS One,
  • Study Type:Method Development Study,
  • Source: PMID: 41325399, DOI: 10.1371/journal.pone.0331101
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