Phylogenomic reassessment of Pseudomonas moraviensis reveals multiple misclassified strains

The taxonomic boundaries of Pseudomonas moraviensis remain unclear despite the increasing availability of genomic data. In this study, we performed a comprehensive polyphasic taxonomic reassessment of 30 publicly available strains, together with 2 newly isolated wheat rhizosphere strains, S9433 and S9602, which exhibited genomic and phenotypic traits indicative of plant growth-promoting potential. Phylogenetic and phylogenomic analysis consistently revealed that strains currently assigned to P. moraviensis are distributed across several distinct lineages. Overall genomic relatedness indices, including average nucleotide identity (ANIb) and digital DNA-DNA hybridization (dDDH), supported the reclassification of multiple strains into different Pseudomonas species. Twenty-two strains including S9433 and S9602 were confirmed as P. moraviensis, whereas five were reclassified as Pseudomonas atacamensis, two as Pseudomonas parakoreensis and one as Pseudomonas iranensis. Strain #71, formerly designated as the type strain of P. moraviensis subsp. stanleyae, formed a distinct phylogenetic lineage with ANIb and dDDH values below species delineation thresholds and a divergent fatty acid profile, collectively indicating that it represents a novel species. Strain VanB exhibited ambiguous genomic relatedness to Pseudomonas aphyarum, Pseudomonas paraglycinae and Pseudomonas fitomaticsae, with ANIb values approaching but dDDH values well below species delineation thresholds, leaving its taxonomic status unresolved pending further investigation. This study clarifies and resolves the taxonomic status of currently assigned P. moraviensis strains and lays a foundation for the functional characterization and applied utilization of this species complex.