The new genus Actinobispora is described. The organisms of this genus are gram positive and aerobic and produce spores in longitudinal pairs on vegetative mycelium and in longitudinal pairs or singly on aerial mycelium. The cell wall contains meso-diaminopimelic acid, and whole-cell hydrolysates contain galactose, arabinose, and xylose (an unusual type IV cell wall); the phospholipid pattern is of type PIV; the major menaquinones are MK-7(H2) and MK-9(H2). No mycolic acids are present. The guanine-plus-cytosine of the DNA is 71.0 mol%. The type species of this genus is Actinobispora yunnunensis. The type strain of this new species is strain Y-11981 (=CCTCC M 90959).During the course of study of the actinomycete population and resources, one actinomycete, strain Y-11981, was isolated from a soil sample from northeast Yunnan, China. Strain Y-11981 belongs to a new genus because of its morphological features and cell composition. In this paper, we describe the morphological, biochemical, and physiological characteristics of the strain, for which we propose the name Actinobispora yunnanensis.
MATERIALS AND METHODSStrain. Strain Y-11981 was isolated from a soil sample collected in Weixin, Yunnan, People's Republic of China. This soil sample had been plated on glycerol-asparagine agar (13) and incubated for 2 weeks at 28°C.Morphology. The media used for micromorphological studies were yeast extract-malt extract agar (ISP 2) and oatmeal agar (ISP 3) (13) incubated for 14 to 21 days at 28°C. Spore morphological observations were made with a light microscope and a model EPMA-8705 electron microscope.Cultural and physiological characteristics. The media and procedures used to determine the cultural and physiological characteristics and carbon and nitrogen source utilization of the strain were those described by Shirling and Gottlieb (13), Locci (9), Furumai et al. (3), Tuite (16), Waksman (17), Gordon (4), and Gordon and Horan (5). Color determinations were made by comparing the cultures with color chips from the ISCC-NBS Color Charts standard sample no. 2106.Whole-cell analysis. The procedures of Becker et al.(1) and Lechevalier (7) were used for whole-cell analysis.Cell wall preparation. Cell walls were purified and analyzed by the method of Lechevalier and Lechevalier (8).Phospholipid analysis. The phospholipid analysis was carried out by the method of Lechevalier and Lechevalier (8).
Menaquinones analysis. Menaquinones were analyzed by the procedures of Collins (2).Guanine-plus-cytosine content. DNA extraction and purification were performed by the methods of Marmur (10) and Zhou (18). To facilitate lysis of the cells by lysozyme, the cells were suspended in IX SSC ( l x SSC is 0.15 M NaCl plus 0.015 M sodium &rate)-1 M sucrose with 10 to 20 mg of lysozyme per ml and incubated for 30 min at 37°C. Three thermal melts were performed with a model DU-7 spectrophotometer. DNA from Escherichia coli K-12 was used as a reference.
RESULTSMicromorphology. Strain Y-11981 was gram positive and not acid fast. The vegetative hyphae...