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Journal of Plant Diseases and Protection

, Volume 126, Issue 6, pp 517–528 | Cite as

Quantitative real-time PCR assay for rapid detection of Pseudomonas amygdali pv. lachrymans in cucumber leaf rinse

  • Filip Gazdik
  • Eliska Penazova
  • Jana Cechova
  • Miroslav Baranek
  • Ales EichmeierEmail author
Original Article
  • 104 Downloads

Abstract

Pseudomonas amygdali pv. lachrymans (Pal) is a bacterium that causes angular leaf spot disease of cucumbers. There is no direct protection against this pathogen, only prevention. Previously published detection systems are labour-intensive and time-consuming. The aim of this study was to create a rapid and reliable diagnostic assay for the detection of Pal in cucumber plants without the need to destroy or damage the plant. The created method is based on washing the bacteria off the leaf surface with saline solution and using the sample directly in a quantitative real-time PCR assay. This detection system is based on the specific amplification of a small coding region of the bacteriophage N4 adsorption protein B gene. The assay was tested on 120 plant samples inoculated with seven different Pal strains. The detection limit of the assay was 2.60 × 102 CFU ml−1, and the concentration of Pal in the leaf rinse ranged from 2.71 × 102 to 6.94 × 107 CFU ml−1 depending on the particular strain. This assay is a reliable and rapid diagnostic method for the detection of Pal from cucumber leaves and is valuable for contributing to angular leaf spot management.

Keywords

Pseudomonas amygdali pv. lachrymans Detection nfrB gene Leaf rinse Real-time PCR 

Notes

Acknowledgements

The work was supported by the Project EFRR “Multidisciplinary research to increase application potential of nanomaterials in agricultural practice” (No. CZ.02.1.01/0.0/0.0/16_025/0007314). This research was also supported by the Project Nos. TJ01000274 and LTC18009.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

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Copyright information

© Deutsche Phytomedizinische Gesellschaft 2019

Authors and Affiliations

  • Filip Gazdik
    • 1
  • Eliska Penazova
    • 1
  • Jana Cechova
    • 1
  • Miroslav Baranek
    • 1
  • Ales Eichmeier
    • 1
    Email author
  1. 1.Mendeleum - Institute of GeneticsMendel University in BrnoLedniceCzech Republic

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