A reliable qPCR technique for detecting viable Xanthomonas arboricola pv. pruni cells.
Xanthomonas arboricola pv. pruni
Cut-off Ct
Intercalating dye
LOD
V-qPCR
Viable cell detection
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
25 Sep 2024
25 Sep 2024
Historique:
received:
17
07
2024
accepted:
15
08
2024
revised:
14
08
2024
medline:
25
9
2024
pubmed:
25
9
2024
entrez:
25
9
2024
Statut:
epublish
Résumé
Xanthomonas arboricola pv. pruni (Xap) is the causal agent of bacterial spot of stone fruits and almond (Prunus spp). Detection of Xap is typically carried out using quantitative real-time PCR (qPCR) combined with culture-based isolation. However, qPCR does not differentiate between viable and dead cells, potentially leading to an overestimation of the infective population in a sample. Such overestimation could result in unnecessary phytosanitary measures. The present study aims to develop a specific protocol ideally targeting to detection of only live Xap bacterial cells. To address this challenge, the viable quantitative PCR (v-qPCR) method was evaluated using three nucleic acid-binding dyes: propidium monoazide (PMA), a combination of PMA and ethidium monoazide (EMA), and PMAxx™, an improved version of PMA. PMAxx™ proved to be the most suitable dye for the detection and quantification of living bacterial cells. This methodology was also evaluated in infected plant material over time and can be considered a rapid and reliable alternative to PCR methods for detecting only those putative infective Xap that may pose a risk for Prunus crops. KEY POINTS: • Protocol to detect biofilm and planktonic viable X. arboricola pv. pruni cells. • Host validated protocol. • Benefits, reduction of chemicals in disease control.
Identifiants
pubmed: 39320527
doi: 10.1007/s00253-024-13288-y
pii: 10.1007/s00253-024-13288-y
doi:
Substances chimiques
Propidium
36015-30-2
Azides
0
propidium monoazide
0
8-azidoethidium
69498-50-6
Types de publication
Journal Article
Evaluation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
472Informations de copyright
© 2024. The Author(s).
Références
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