Two Gram-negative, non-motile, rod-shaped strains, BQ4-1 T and NHI3-2, isolated respectively from the healthy and diseased part of Populus ¾euramericana canker bark, were characterized using a polyphasic approach. Chemotaxonomic characterization supported the inclusion of the two strains in the genus Acinetobacter, with genomic DNA G+C contents (42.5-43 mol%) within the range observed for this genus (38-47 mol%) and 9-octadecenoic acid (C 18 : 1 v9c, 39.87 %), hexadecanoic acid (C 16 : 0 , 11.26 %) and summed feature 3 (comprising C 16 : 1 v7c/ C 16 : 1 v6c, 18.90 %) as major fatty acids. Phylogenetic analysis based on 16S rRNA, rpoB and gyrB gene sequences revealed that strains BQ4-1 T and NHI3 did not cluster with any species with validly published names, and formed a distinct cluster with 99-100 % bootstrap support on three phylogenetic trees within the genus Acinetobacter. Acid was not produced from D-glucose, and haemolysis was not observed on agar media supplemented with sheep erythrocytes. On the basis of phenotypic, genotypic and phylogenetic characteristics, the two strains are considered to represent a novel species of the genus Acinetobacter, for which the name Acinetobacter puyangensis sp. nov. is proposed. The type strain is BQ4-1 T (5CFCC 10780 T 5JCM 18011 T ).