Strain HUAS CB01(T) was a novel actinobacterium which was isolated from the rhizosphere soil of Cathaya argyrophylla, Chengbu Miao Autonomous County of Hunan Province, China. The strain formed well-growing substrate mycelium, diffusible pigments, and aerial mycelium, and differentiated into spiral-type spore chains composed of smooth-surface rod-shaped spores. Phylogenetic analysis on account of 16 S rRNA gene sequence demonstrated the strain HUAS CB01(T) was a member of the genus Streptomyces and had a close relationship with Streptomyces wuyuanensis CGMCC 4.7042 (T) (100%) and Streptomyces marianii ICN19(T) (99.86%). Genome-based comparisons indicated that strain HUAS CB01(T) could be distinctly different from its closest species, Streptomyces wuyuanensis CGMCC 4.7042 (T), Streptomyces marianii ICN19(T), with ANIm and dDDH results of 92.78% and 45.90%, 92.22% and 43.30%, respectively, far less than 96.7 and 70% cut-off points recommended for delineating species. The main cellular fatty acids concluded anteiso-C-15:0, iso-C-14:0, iso-C-16:0, C-16:0 and C-16:1 2OH. The menaquinones were MK-9(H-4), MK-9(H-6) and MK-9(H-8) and the whole-cell sugars consisted of ribose and mannose. The polar lipids included phosphatidyl ethanolamine, diphosphatidylglycerol, phosphatidylglycerol, mannosides and unidentified phospholipids. According to these genotypic and phenotypic characteristics, strain HUAS CB01(T) can be distinguished and representative to be a novel species of the genus Streptomyces, for which the name Streptomyces chengbuensis is proposed. The type strain is HUAS CB01(T) ( = MCCC 1K08666(T) = JCM 36277 (T)).