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Lookup NU author(s): Dr Emanuela Torelli
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2024 The Author(s). Plant Pathology published by John Wiley & Sons Ltd on behalf of British Society for Plant Pathology.Xylophilus ampelinus is a xylematic bacterium causing bacterial blight of grapevine, a disease regarded as a potential threat for viticulture in several countries. Currently, PCR detection is pivotal in diagnostic protocols due to the bacterium's infrequent occurrence in the field and the technical advantages of PCR. Recent metagenomic studies have unveiled diversity in its taxonomic domain, unknown when the most widely used assays for the detection of X. ampelinus infections were developed. In particular, PCR assays relying on highly conserved sequence regions, such as those surrounding ribosomal RNA genes, may be substituted with more specific PCR assays. In this study, we first investigated the diversity of detectable grapevine endophytes related to but different from X. ampelinus and delineated the genotaxonomic boundaries of the species in relation to the known (meta)genomes of closely related bacteria. Then, by exploiting the wealth of genomes now available for bacteria classified in the Burkholderiales, we devised several sets of primers targeting only X. ampelinus and its closest relatives. These primers were employed to (i) genotaxonomically circumscribe the grapevine endophyte species related to but distinguishable from X. ampelinus and (ii) develop a robust multiplex PCR assay expected to be specific for the species X. ampelinus based on in vitro and in silico evidence. The adoption of the multiplex PCR assay presented here is expected to reduce the risk of false positives in the diagnosis of bacterial blight of grapevine.
Author(s): Carminati G, Bianchi GL, DeAmicis F, Cannistraci I, Sadallah A, Ermacora P, Torelli E, Martini M, Firrao G
Publication type: Article
Publication status: Published
Journal: Plant Pathology
Year: 2024
Pages: epub ahead of print
Online publication date: 03/10/2024
Acceptance date: 06/09/2024
Date deposited: 14/10/2024
ISSN (print): 0032-0862
ISSN (electronic): 1365-3059
Publisher: John Wiley and Sons Inc
URL: https://doi.org/10.1111/ppa.14013
DOI: 10.1111/ppa.14013
Data Access Statement: Data generated in this study are available in part as article supplementary material and in part in the European Nucleotide Archive at www.ebi.ac.uk/ena/ as project PRJEB75643.
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