Six Multiplex TaqMan-qPCR Assays for Quantitative Diagnostics of Pseudomonas Species Causative of Bacterial Blotch Diseases of Mushrooms

Author: mycolabadmin
2020-05-25
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Summary

This research developed new molecular testing methods to detect and measure harmful bacteria that cause diseases in cultivated mushrooms. The tests are more accurate and sensitive than previous methods, allowing early detection of pathogens before they can cause significant crop damage. Impact on everyday life: – More reliable supply and quality of mushrooms for consumers – Reduced food waste from diseased mushroom crops – Lower production costs that could lead to more affordable mushrooms – Better food safety through early pathogen detection – More sustainable mushroom farming practices through targeted disease control

Background

Bacterial blotch is a group of economically important diseases of the common button mushroom (Agaricus bisporus). The disease can reduce crop yields by up to 50% in Europe alone. Once pathogens are introduced to a farm, the mesophilic growing conditions optimal for mushroom production result in severe and widespread secondary infections. Efficient detection of pathogens is critical for disease control.

Objective

To develop real-time TaqMan assays for molecular diagnosis of three currently prevalent bacterial blotch pathogens: Pseudomonas gingeri, Pseudomonas tolaasii and uncharacterized Pseudomonas strains belonging to P. salomonii and P. edaphica. The assays aim to enable quantitative detection of these pathogens in bacterial cultures, symptomatic tissue, infected casing soil and water samples from mushroom farms.

Results

All six developed assays demonstrated high diagnostic specificity and sensitivity when tested against the bacterial panel. The assays showed good analytical performance with linearity across calibration curves (>0.95), amplification efficiency (>90%) and strong amplification signals (>2.1). The detection limits were optimized for efficient quantification in various sample types. The multiplexed assays successfully detected and quantified pathogen populations in environmental samples from mushroom farms.

Conclusion

The developed TaqMan assays provide reliable tools for quick and quantitative detection of three aggressive bacterial blotch pathogens in different substrates associated with mushroom cultivation. The assays enable efficient diagnosis of secondary infections and can assess the effectiveness of disinfectants and biocontrol agents. They represent the first opportunity to accurately quantify pathogen populations in environmental samples and study their microbial ecology and population dynamics.

Published in:Frontiers in Microbiology,
Study Type:Methods Development,
Source: 10.3389/fmicb.2020.00989