During the course of our research on new actinobacterial sources, a novel actinomycete strain YIM 63101(T) was isolated from the surface-sterilized roots of Artemisia annua L. collected from Xishuangbanna, Yunnan province, south-west China and characterized by using a polyphasic approach. The strain formed well-differentiated aerial and substrate mycelia. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 63101(T) belongs to the genus Pseudonocardia, with highest similarity to "Pseudonocardia artemisiae YIM 63587(T)" (99.4%). Sequence similarities between strain YIM 63101(T) and the other Pseudonocardia species ranged from 97.0 (Pseudonocardia saturnea IMSNU 20052(T)) to 94.0% (Pseudonocardia compacta IMSNU 20111(T)). The chemotaxonomic characteristics, such as cell wall diaminopimelic acid, whole-cell sugars, fatty acid components and the major menaquinones suggested that the organism belonged to the genus Pseudonocardia. The G + C content of the genomic DNA was 69.4 mol%. Based on comparative analysis of physiological, biochemical and chemotaxonomic data, including low DNA-DNA hybridization results, it is proposed that strain YIM 63101(T) represents a novel species of the genus Pseudonocardia, named Pseudonocardia bannaensis sp. nov. The type strain is YIM 63101(T) (= CCTCC AA 208077 (T) = DSM 45300(T)).