Background and aims <p>Eucalyptus has unique wood properties and great economic value, but traditional yield-increasing methods rely on chemicals and harm the environment. Plant growth-promoting rhizobacteria (PGPR) are a good solution to this problem, and <i>Pseudomonas</i> is a typical example. The study aims to explore sustainable, eco-friendly alternatives to minimize chemical pollution of the environment by screening and characterizing <i>Pseudomonas</i> with the potential to promote the growth of eucalyptus.</p> Methods <p>The Salkowski colorimetric method and various selective media were used to screen for bacteria with growth-promoting functions. Genes associated with growth-promoting functions were identified by genome annotation. A combination of phylogenetic, comparative genomic, morphological, physiological and biochemical methods was used to classify and identify the strains.</p> Results <p>Genomic annotation identified genes associated with growth-promoting functions (e.g., <i>ppk</i> for phosphate solubilization, <i>trpA</i>/<i>B</i>/<i>C</i> for IAA synthesis), consistent with the results of plate experiments, further confirming their growth-promoting potential. The two strains were classified as new species within the <i>Pseudomonas</i> genus, and named <i>Pseudomonas eucalyptica</i> sp. nov. and <i>Pseudomonas eucalyptoterra</i> sp. nov.</p> Conclusion <p>This study provides new ideas for the development of green, non-polluting biofertilizers and adds to our knowledge of <i>Pseudomonas</i> diversity.</p>