Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Sat, 15 Dec 2018 05:15:52 Herbidospora osyris sp. nov., isolated from surface- sterilized tissue of Osyris wightiana Wall. ex Wight Jie Li, 1 Guo-Zhen Zhao, 1 Sheng Qin, 1 Wen-Yong Zhu, 1 Li-Hua Xu 1 and Wen-Jun Li 1,2 Correspondence Wen-Jun Li wjli@ynu.edu.cn or liact@hotmail.com 1 The Key Laboratory for Microbial Resources of the Ministry of Education, PR China, and Laboratory for Conservation and Utilization of Bio-Resources, Yunnan Institute of Microbiology, Yunnan University, Kunming 650091, PR China 2 Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, PR China An endophytic actinomycete, strain YIM 65070 T , was isolated from surface-sterilized tissue of Osyris wightiana Wall. ex Wight collected from Yunnan province, south-west China, and characterized by using a polyphasic approach. Strain YIM 65070 T had morphological and chemotaxonomic markers that were consistent with its classification in the genus Herbidospora. Phylogenetic analysis based on almost complete 16S rRNA gene sequences indicated that strain YIM 65070 T was phylogenetically very closely related to Herbidospora cretacea IFO 15474 T . DNA–DNA hybridization experiments confirmed the separate genomic status of strains YIM 65070 T and H. cretacea DSM 44071 T . Moreover, strain YIM 65070 T could be distinguished from H. cretacea DSM 44071 T by differences in several phenotypic characteristics such as tolerance to NaCl, degradation activity, utilization of sole carbon and nitrogen sources and the cellular fatty acid contents. On the basis of phenotypic and phylogenetic evidence, strain YIM 65070 T was identified as a novel species of the genus Herbidospora, for which the name Herbidospora osyris sp. nov. is proposed, with YIM 65070 T (5CCTCC AA 208019 T 5DSM 45214 T ) as the type strain. The genus Herbidospora of the family Streptosporangiaceae was first described by Kudo et al. (1993). At the time of writing, the genus comprises only one recognized species, Herbidospora cretacea (Kudo et al., 1993). The members of this genus produce branching substrate mycelia, but no distinct aerial hyphae. Short chains of non-motile spores (10–30 spores per chain) are borne on the tips of sporophores, which are derived from the vegetative mycelia in clusters. Cell walls contain meso-diaminopimelic acid and N-acetylated muramic acid, but lack a significant amount of glycine. Whole-cell hydrolysates contain a trace amount of madurose. The phospholipid pattern is the type PIV pattern of Lechevalier et al. (1977). The cellular fatty acid composition is characterized by the presence of iso- hexadecanoic, n-hexadecanoic, n-heptadecenoic, 10- methyl heptadecanoic and 2-hydroxy acids. The predom- inant menaquinone for the genus is MK-10(H 4 ) and MK- 10(H 6 ), MK-10(H 2 ), MK-10(H 0 ) and MK-9(H 4 ) are also present as minor components. The G+C contents of the genomic DNA are 69–71 mol%. Strain YIM 65070 T was isolated as part of a discovery and identification programme for endophytic actinomycetes from plants in Yunnan province, south-west China. Strain YIM 65070 T was isolated from surface-sterilized tissue of Osyris wightiana Wall. ex Wight. The surface sterilization and isolation procedures were performed according to previously described methods (Li et al., 2009). The purified strain was routinely cultured on YIM 38 medium [4 g malt extract, 4 g yeast extract, 4 g glucose, vitamin mixture (0.25 mg biotin and 0.5 mg each of p-aminobenzoic acid, calcium pantothenate, inositol, niacin, pyridoxine-HCl, riboflavin and thiamine-HCl), 20 g agar; pH 7.2] at 28 u C and stored as a glycerol suspension (20 %, v/v) at 270 u C. Genomic DNA extraction, amplification and 16S rRNA gene sequencing were performed as described previously by Li et al. (2007). An almost complete 16S rRNA gene sequence of strain YIM 65070 T , comprising 1425 bp, was obtained and aligned with corresponding sequences of H. cretacea and other members of the family Strepto- sporangiaceae (retrieved from the GenBank/EMBL/DDBJ database) using CLUSTAL_X (Thompson et al., 1997). The resulting alignment was corrected manually and a phylo- genetic tree was constructed using the neighbour-joining (Saitou & Nei, 1987) tree-making algorithm from MEGA The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain YIM 65070 T is FJ214356. International Journal of Systematic and Evolutionary Microbiology (2009), 59, 3123–3127 DOI 10.1099/ijs.0.010579-0 010579 G 2009 IUMS Printed in Great Britain 3123