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Real-Time PCR Assays for the Detection of Puccinia psidii

J Baskarathevan1, R K Taylor1, W Ho1

  • 1Plant Health and Environment Laboratory, Ministry for Primary Industries, Auckland 1140, New Zealand.

Plant Disease
|January 29, 2019
PubMed

Insights

Novel qPCR assays detect Myrtle rust (Puccinia psidii) in plants. These highly specific and sensitive methods enable early detection, crucial for managing this significant global threat to the Myrtaceae family.

Area of Science:

  • Plant Pathology
  • Molecular Biology
  • Genetics

Background:

  • Myrtle rust (Puccinia psidii) is an emerging pathogen threatening the global Myrtaceae family.
  • Increasing geographic range and broad host susceptibility pose significant risks worldwide.

Purpose of the Study:

  • Develop and validate novel real-time quantitative PCR (qPCR) assays for P. psidii detection.
  • Enhance early and high-throughput detection capabilities for effective disease management.

Main Methods:

  • Designed three qPCR assays targeting ribosomal DNA and β-tubulin gene sequences.
  • Validated assays using DNA from urediniospores and infected plant tissues, including asymptomatic samples.
  • Developed a duplex assay with an internal host control (cytochrome oxidase gene).

Main Results:

  • All qPCR assays successfully detected P. psidii DNA, with sensitivity down to 0.011 pg.
  • The optimal assay demonstrated high specificity, repeatability, and reproducibility across platforms and labs.
  • Duplex assay provided an internal control for reliable results.

Conclusions:

  • Developed rapid, sensitive, and specific qPCR assays for P. psidii detection.
  • These assays facilitate early infection identification before sporulation, aiding disease surveillance.
  • Optimized qPCR offers a superior alternative to previous methods for high-throughput testing.

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