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Detection and Identification of Six Monilinia spp. Causing Brown Rot Using TaqMan Real-Time PCR from Pure Cultures

Jing-Ru Wang1, Li-Yun Guo1, Chang-Lin Xiao2

  • 1Department of Plant Pathology and MOA Key Lab of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, 100193, Beijing, China.

Plant Disease
|January 24, 2019
PubMed

Insights

New real-time PCR assays accurately detect and identify six brown rot species affecting fruit crops. These sensitive molecular tools offer rapid and specific pathogen identification, crucial for disease management and quarantine efforts.

Area of Science:

  • Plant Pathology
  • Molecular Biology
  • Agricultural Science

Background:

  • Brown rot is a significant disease impacting stone and pome fruits globally.
  • Six closely related species (Monilinia fructicola, M. laxa, M. fructigena, Monilia polystroma, M. mumecola, and M. yunnanensis) are confirmed causal agents.
  • Geographic distribution differences make some species critical quarantine pathogens.

Purpose of the Study:

  • To develop and validate TaqMan real-time polymerase chain reaction (PCR) assays for the precise detection and identification of six brown rot Monilinia species.
  • To establish sensitive and specific molecular diagnostic tools for these important plant pathogens.

Main Methods:

  • Design of species-specific primer pairs and probes targeting laccase-2 genes for M. fructicola, M. fructigena, M. laxa, and M. polystroma.
  • Design of primer pairs and probes targeting elongation factor genes for M. mumecola and M. yunnanensis.
  • Validation of real-time PCR assays using purified DNA, mixed samples, and infected apple fruit.

Main Results:

  • Developed highly specific TaqMan real-time PCR assays capable of identifying the six target Monilinia species.
  • Achieved high sensitivity with detection limits ranging from 10 to 100 fg of DNA (equivalent to 1–7 conidia).
  • Successfully detected target pathogens in mixed DNA samples and infected apple fruit, with results consistent with morphological identification.

Conclusions:

  • The developed real-time PCR assays provide a rapid, sensitive, and specific method for identifying key brown rot pathogens.
  • These molecular tools are valuable for accurate diagnosis, disease management, and enforcing quarantine regulations in agriculture.
  • The assays offer an improvement over traditional methods for brown rot species identification in affected plant tissues.

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