The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Development of the Droplet Digital PCR Method for the Detection and Quantification of Erwinia pyrifoliae

  • Lin, He (Department of Applied Biology, Chungnam National University) ;
  • Seong Hwan, Kim (Department of Microbiology, Dankook University) ;
  • Jun Myoung, Yu (Department of Applied Biology, Chungnam National University)
  • Received : 2022.08.23
  • Accepted : 2023.01.10
  • Published : 2023.02.01
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Abstract

Black shoot blight disease caused by Erwinia pyrifoliae has serious impacts on quality and yield in pear production in Korea; therefore, rapid and accurate methods for its detection are needed. However, traditional detection methods require a great deal of time and fail to achieve absolute quantification. In the present study, we developed a droplet digital polymerase chain reaction (ddPCR) method for the detection and absolute quantification of E. pyrifoliae using a pair of species-specific primers. The detection range was 103-107 copies/ml (DNA templates) and cfu/ml (cell culture templates). This new method exhibited good linearity and repeatability and was validated by absolute quantification of E. pyrifoliae DNA copies from samples of artificially inoculated immature pear fruits. Here, we present the first study of ddPCR assay for the detection and quantification of E. pyrifoliae. This method has potential applications in epidemiology and for the early prediction of black shoot blight outbreaks.

Keywords

Acknowledgement

We thank Dr. Sunghoon Jung for the use of laboratory equipment, and Dr. Mi-Hyun Lee for providing E. amylovora genomic DNA. This project was supported by the Animal and Plant Quarantine Agency (PQ20204B026), Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

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