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One-Step Detection of Monilinia fructicola, M. fructigena, and M. laxa on Prunus and Malus by a Multiplex Real-Time

C Guinet1, C Fourrier-Jeandel1, I Cerf-Wendling1

  • 1Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail (ANSES) Laboratoire de la Santé des Végétaux, Unité de Mycologie, Domaine de Pixérécourt, 54220 Malzéville, France.

Plant Disease
|January 29, 2019
PubMed

Insights

A new multiplex real-time PCR assay effectively detects three key brown rot fungi (Monilinia spp.) in fruit and orchard samples. This sensitive and specific tool aids in accurate disease diagnosis for regulatory control.

Area of Science:

  • Plant Pathology
  • Molecular Biology
  • Agricultural Science

Background:

  • Brown rot is a significant fungal disease impacting fruit production and trade.
  • Monilinia fructicola, M. laxa, and M. fructigena are the primary causal agents, with varying regulatory statuses.
  • Accurate detection is crucial for disease management and regulatory compliance.

Purpose of the Study:

  • To develop a novel multiplex real-time polymerase chain reaction (PCR) assay.
  • To enable simultaneous detection of three Monilinia species in a single reaction.
  • To provide a reliable diagnostic tool for fruit and plant tissues.

Main Methods:

  • Development of species-specific primer-hydrolysis probe combinations for Monilinia spp.
  • Utilizing a MO368 characterized amplified region marker.
  • Incorporation of a universal 18S Uni primer-probe test for DNA quality control.
  • Quadruplex real-time PCR format for multiplex detection.

Main Results:

  • The developed multiplex assay demonstrated high sensitivity, specificity, reproducibility, and robustness.
  • The assay successfully detected the three target Monilinia species in fruit, twigs, and flowers of Prunus and Malus spp.
  • Implemented quality controls minimized the risk of false-positive and false-negative results.

Conclusions:

  • The new multiplex real-time PCR assay is a high-performance tool for brown rot detection.
  • The assay is suitable for serial analyses and reliable for official controls in fruit production.
  • This method enhances diagnostic capabilities for managing economically important fungal diseases.

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