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Quinoa is an expanding crop in southern Spain. Downy mildew, caused by Peronospora variabilis, is the most important quinoa disease in Spain and worldwide. In Spain, this disease has also been observed on the weed Chenopodium album. The objectives of this study were to unravel the origin of the P. variabilis isolates currently infecting quinoa in southern Spain and to study their genetic diversity. We hypothesized that P. variabilis isolates infecting quinoa in Spain could have been introduced through the seeds of the quinoa varieties currently grown in the country or, alternatively, that these isolates are endemic isolates, originally infecting C. album, that jumped to quinoa. In order to test these hypotheses, we sequenced the internal transcribed spacer (ITS), cytochrome c oxidase subunit 1 (cox1), and cox2 regions of 33 P. variabilis isolates infecting C. quinoa and C. album in southern Spain and analyzed their phylogenetic relationship with isolates present in other countries infecting Chenopodium spp. cox1 gene sequences from all of the Spanish P. variabilis isolates were identical and exhibited nine single-nucleotide polymorphisms (SNPs) compared with a single P. variabilis cox1 sequence found at GenBank. Phylogenetic analyses based on the ITS ribosomal DNA region were not suitable to differentiate isolates according to their geographical origin or host. The cox2 sequences from P. variabilis Spanish isolates collected from C. quinoa and C. album were all identical and had a distinctive SNP in the last of four polymorphic sites that distinguished Spanish isolates from isolates from other countries. These results suggest that P. variabilis infecting quinoa in southern Spain could be native isolates that originally infected C. album. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

This work was funded by the United States Department of Agriculture–Agricultural Research Service appropriated project 2034-22000-012-00D and the Xylella fastidiosa Active Containment Through a Multidisciplinary-Oriented Research Strategy (XF-ACTORS) project. Illumina sequencing for BB08-1 was carried out at the University of California, Davis, Genome Center DNA Technologies and Expression Analysis Cores, supported by National Institute of Health Shared Instrumentation Grant 1S10OD010786-01.

The plant pathogenic bacterium Xylella fastidiosa (Xf) is a major threat to agriculture and the environment worldwide. Recent devastating outbreaks in Europe highlight the potential of this pathogen to cause emergent diseases. Xf subsp. multiplex ESVL and IVIA5901 strains that belong to sequence type (ST) 6 were isolated from almond orchards within the outbreak area in Alicante province (Spain). Both strains share more than 99% of the chromosomal sequences), but the ESVL strain harbours two plasmids (pXF64-Hb_ESVL and pUCLA-ESVL). Here, virulence phenotypes and genome content were compared between both strains, using three strains from USA as a reference for the phenotypic analyses. Experiments in microfluidic chambers, used as simulation of xylem vessels, showed that twitching motility was absent in the IVIA5901 strain, while the ESVL strain had reduced twitching motility. Both Spanish strains had less biofilm formation, cell aggregation and lower virulence in tobacco compared with USA reference strains. Genome analysis of the two plasmids from ESVL revealed 51 unique coding sequences (CDS) that were absent in the chromosome of IVIA5901. Comparison of the chromosomes of both strains showed some unique CDS and SNPs in each strain with potential deleterious mutations. Genomic differences found in genes previously associated with adhesion and motility might explain the differences in the phenotypic traits studied. Although additional studies are necessary to infer the potential role of Xf plasmids, our results indicate that the presence of plasmids should be considered in the study of the mechanisms of pathogenicity and adaptation in Xf to new environments.

The plant pathogenic bacterium Xylella fastidiosa (Xf) is a major threat to agriculture and the environment worldwide. Recent devastating outbreaks in Europe highlight the potential of this pathogen to cause emergent diseases. Xf subsp. multiplex ESVL and IVIA5901 strains that belong to sequence type (ST) 6 were isolated from almond orchards within the outbreak area in Alicante province (Spain). Both strains share more than 99% of the chromosomal sequences), but the ESVL strain harbours two plasmids (pXF64-Hb_ESVL and pUCLA-ESVL). Here, virulence phenotypes and genome content were compared between both strains, using three strains from USA as a reference for the phenotypic analyses. Experiments in microfluidic chambers, used as simulation of xylem vessels, showed that twitching motility was absent in the IVIA5901 strain, while the ESVL strain had reduced twitching motility. Both Spanish strains had less biofilm formation, cell aggregation and lower virulence in tobacco compared with USA reference strains. Genome analysis of the two plasmids from ESVL revealed 51 unique coding sequences (CDS) that were absent in the chromosome of IVIA5901. Comparison of the chromosomes of both strains showed some unique CDS and SNPs in each strain with potential deleterious mutations. Genomic differences found in genes previously associated with adhesion and motility might explain the differences in the phenotypic traits studied. Although additional studies are necessary to infer the potential role of Xf plasmids, our results indicate that the presence of plasmids should be considered in the study of the mechanisms of pathogenicity and adaptation in Xf to new environments.