This paper describes two new species of actinomycetes in the genus Catellatospora, isolated from woodland soils in Japan. Catellatospora matsumotoense and C. tsunoense were identified based on physiological, morphological, and chemotaxonomic characteristics that distinguished them from previously described Catellatospora species. The two new species differ in their menaquinone composition (MK-10 versus MK-9), presence or absence of 3-O-methylrhamnose in cell walls, novobiocin susceptibility, vitamin requirements, and carbohydrate utilization patterns. DNA-DNA hybridization confirmed their status as distinct species with low homology to other Catellatospora species. C. matsumotoense contains MK-10, is novobiocin-resistant, does not require thiamine, and utilizes fructose and raffinose but not mannitol. C. tsunoense contains MK-9, is novobiocin-susceptible, requires thiamine, and utilizes fructose, raffinose, and rhamnose. Both species have similar DNA G+C contents (around 70-71 mol%) and were recovered exclusively from woodland soils across Japan, suggesting the genus is widespread in temperate forest environments.

Key findings

• Two new Catellatospora species were identified and formally described: C. matsumotoense (type strain 6393-C) and C. tsunoense (type strain 6420-P), distinguished by menaquinone type, cell wall composition, and metabolic profiles
• C. matsumotoense (MK-10) is novobiocin-resistant and does not require thiamine, while C. tsunoense (MK-9) is novobiocin-susceptible and requires thiamine for growth
• The two species show differential utilization of specific carbohydrates including fructose, raffinose, mannitol, and rhamnose, providing useful diagnostic characteristics
• DNA-DNA hybridization demonstrated low homology between the new species and previously described Catellatospora species, confirming their distinct taxonomic status
• All Catellatospora isolates were recovered exclusively from woodland soils across Japan, suggesting ecological preference for temperate forest environments