Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR

Delgado Santander, Ricardo; Gašić, Katarina; Aćimović, Srđan

doi:10.1007/978-1-0716-2517-0_14

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Delgado Santander, Ricardo
Gašić, Katarina

Aćimović, Srđan

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Abstract

The accurate assessment of Erwinia amylovora live cell populations in fire blight cankers by classic microbiology methods has major limitations. Some of them are the presence of competitive microbiota in samples that inhibit E. amylovora’s growth and the release of toxic compounds by plant material during sample processing, which may hamper the pathogen’s ability to form colonies on solid media. Digital PCR (dPCR) combined with the photo-reactive DNA-binding dye propidium monoazide (PMA) allows selective detection and quantification of live E. amylovora cells in woody samples while overcoming the constraints of culture-dependent methods. This work describes a reliable viability dPCR procedure to determine E. amylovora live cell concentrations in fire blight cankers from pome fruit trees. This protocol can be adapted for the analysis of other types of plant material and enables investigation of ecological, epidemiological, and management significance of cankers as a relatively underexpl...

Keywords:

Fire blight / Cankers / Bacterial survival / Bacterial viability / Propidium monoazide / dPCR / Molecular detection methods / Apple / Pear / Asian pear

Source:
Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536), 2022, 231-249

Publisher:

Springer

Funding / projects:

National Institute of Food and Agriculture, U.S. Department of Agriculture, Hatch/Multistate Research Program projects NYG-625835 and NYG-625857 under 1009897 and 1014444
NY State Farm Viability Institute grant number 81927/A001-FVI 17 006
NY State Specialty Crop Block Grant Program grant number SCG 82535/A001-SCG 17 005 to SGA
Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200010 (Institute for Plant Protection and Environment, Belgrade) (RS-200010)

DOI: 10.1007/978-1-0716-2517-0_14

ISBN: 9781071625163

ISSN: 1064-3745

Scopus: 2-s2.0-85134427071

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	1

TY - CHAP
AU - Delgado Santander, Ricardo
AU - Gašić, Katarina
AU - Aćimović, Srđan
PY - 2022
UR - https://plantarum.izbis.bg.ac.rs/handle/123456789/765
AB - The accurate assessment of Erwinia amylovora live cell populations in fire blight cankers by classic microbiology methods has major limitations. Some of them are the presence of competitive microbiota in samples that inhibit E. amylovora’s growth and the release of toxic compounds by plant material during sample processing, which may hamper the pathogen’s ability to form colonies on solid media. Digital PCR (dPCR) combined with the photo-reactive DNA-binding dye propidium monoazide (PMA) allows selective detection and quantification of live E. amylovora cells in woody samples while overcoming the constraints of culture-dependent methods. This work describes a reliable viability dPCR procedure to determine E. amylovora live cell concentrations in fire blight cankers from pome fruit trees. This protocol can be adapted for the analysis of other types of plant material and enables investigation of ecological, epidemiological, and management significance of cankers as a relatively underexplored part of the fire blight disease cycle.
PB - Springer
T2 - Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536)
T1 - Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR
EP - 249
SP - 231
DO - 10.1007/978-1-0716-2517-0_14
ER -

@inbook{
author = "Delgado Santander, Ricardo and Gašić, Katarina and Aćimović, Srđan",
year = "2022",
abstract = "The accurate assessment of Erwinia amylovora live cell populations in fire blight cankers by classic microbiology methods has major limitations. Some of them are the presence of competitive microbiota in samples that inhibit E. amylovora’s growth and the release of toxic compounds by plant material during sample processing, which may hamper the pathogen’s ability to form colonies on solid media. Digital PCR (dPCR) combined with the photo-reactive DNA-binding dye propidium monoazide (PMA) allows selective detection and quantification of live E. amylovora cells in woody samples while overcoming the constraints of culture-dependent methods. This work describes a reliable viability dPCR procedure to determine E. amylovora live cell concentrations in fire blight cankers from pome fruit trees. This protocol can be adapted for the analysis of other types of plant material and enables investigation of ecological, epidemiological, and management significance of cankers as a relatively underexplored part of the fire blight disease cycle.",
publisher = "Springer",
journal = "Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536)",
booktitle = "Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR",
pages = "249-231",
doi = "10.1007/978-1-0716-2517-0_14"
}

Delgado Santander, R., Gašić, K.,& Aćimović, S.. (2022). Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR. in Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536)
Springer., 231-249.
https://doi.org/10.1007/978-1-0716-2517-0_14

Delgado Santander R, Gašić K, Aćimović S. Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR. in Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536). 2022;:231-249.
doi:10.1007/978-1-0716-2517-0_14 .

Delgado Santander, Ricardo, Gašić, Katarina, Aćimović, Srđan, "Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR" in Plant Pathology, Part of the Methods in Molecular Biology book series (MIMB,volume 2536) (2022):231-249,
https://doi.org/10.1007/978-1-0716-2517-0_14 . .