An actinobacterial strain, 2SbT, isolated from lichen was characterized taxonomically using a polyphasic approach. Strain 2SbT was Gram-positive, strictly aerobic, rod-shaped and non-motile. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain 2SbT was located in the genus Leifsonia. Level of 16S rRNA gene sequence similarity between the isolated strains and the type strains of Leifsonia species were 95.0-99.2%. The value of DNA-DNA relatedness between strain 2SbT and Leifsonia poae, a phylogenetically related and phenotypically similar species, was 39.9/25.2%. The DNA G+C content of the strain 2SbT was 61.7 mol%. Cell wall peptidoglycan type (2,4-diaminobutyric acid), major cellular fatty acids (anteiso-C(15:0), anteiso-C(17:0) and iso-C(16:0)) and quinone type (MK-11 and MK-10) of the isolate support their affiliation to the genus Leifsonia. On the basis of phylogenetic position, physiological and chemotaxonomic properties, strain 2SbT represent a novel species of the genus Leifsonia, for which the name Leifsonia lichenia sp. nov. is proposed. The type strain is 2SbT (=IAM 15426T =JCM 23226T =KCTC 13122T).