g., [11, 18–24]), which have revealed the diversity and complexity of the genus [23, HSP inhibitor drugs 24], while showing the limitations of single locus analyses [25]. However,

during the last decade the taxonomy of this genus has still been subject to considerable debate. Genus-wide reclassifications have been proposed [26, 27], and frequent sub-specific reclassifications and proposals for new species have been published [19–21, 28–30]. A remarkable example of these conflicts is the classification of X. fuscans aurantifolii [26, 27], also known as X. axonopodis pv. “”aurantifolii”" [2, 6, 18, 31]. This taxon was originally identified as part of the DNA hybridization homology group “”X. axonopodis”" [6], but after its differentiation from other xanthomonads by DNA sequence-based molecular techniques, production of water-soluble brown pigment and host range, it was designated as X. fuscans [26]. However, when these traits/methods were examined, none of them could individually differentiate X. fuscans from other pathovars within X. axonopodis [18, 31]. DNA-DNA reassociation assays, in turn, have differentiated X. fuscans from X. axonopodis, X. campestris and X. citri [2, 26, 27]. Additional host-range evidence has also been used to support the designation X. fuscans, separated from X. axonopodis and X. citri. Phaseolus vulgaris ASK inhibitor and Citrus spp. are infected by X. fuscans pvs. fuscans and aurantifolii,

respectively, but are not infected by either X. axonopodis or X. campestris. Citrus spp., on the other hand, is also infected by X. citri [1]. However, host range is usually a criterion to separate pathovars and not

species. This example underscores the importance of a solid taxonomic classification with a phylogenetic basis. Molecular phylogenetics has this website played an important role in the classification of the genus. Single locus Cyclin-dependent kinase 3 analyses, including the use of 16S-23S rDNA spacers, the 16S rRNA gene and the DNA gyrase gyrB [32–35], generally agree with standing nomenclature but with low resolution below the species level. MLSA including sequences of protein-coding genes dnaK, fyuA and rpoD [31], has significantly extended previous results. In general, MLSA results suggest that X. citri and X. fuscans are closely related species and should be considered as a single species based on their 98.34% similarity in the proteins encoded by dnaK, fyuA, gyrB and rpoD [31]. Recently, a phylogenomic approach was applied to resolve the phylogenetic relationships within the genus [11], although this work did not explore the phylogenetic distances between strains, and did not include sequences from X. axonopodis species. The general structure of the genus agreed with the standing nomenclature. The use of genomic sequences as the basis for species delimitation has been explored as a new standard in bacteria in replacement of DNA-DNA hybridization [36, 37], particularly based on metrics such as the ANI (Average Nucleotide Identity) [38].