Rhizohabitans arisaemae gen. nov., sp. nov., a novel actinomycete of the family Streptosporangiaceae.

An actinomycete strain K14-0274T was isolated from the root of Arisaema thunbergii Blume subsp. urashima (H. Hara) H. Ohashi et J. Murata collected in Japan. The results of phylogenetic analysis based on the 16S rRNA gene sequence indicated thatK14-0274T could be distinguished from the members of all known genera, although it represented a member of the family Streptosporangiaceae. K14-0274T produced sporangium-like spherical vesicles with spores on white aerial mycelia. MK-9 (H4) and MK-9 (H6) were the major menaquinones. The whole-cell hydrolysates contained madurose, glucose, mannose, rhamnose and ribose. The cell-wall amino acids comprise l-alanine, d-alanine, d-glutamic acid and meso-diaminopimelic acid. The N-acyl type of muramic acid was acetyl. Mycolic acids were not detected. Phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside were detected. The predominant fatty acids were iso-C16 : 0, 10-methyl-C18 : 0 and C16 : 0. The G+C content of the genomic DNA was 69.7 mol%. On the basis of morphological, phylogenetic and chemotaxonomic characteristics, strain K14-0427T represents a novel genus in the family Streptosporangiaceae, for which the name Rhizohabitans arisaemae gen. nov., sp. nov. is proposed. The type strain is K14-0247T (=NBRC 114594T =TBRC 12948T).

Amycolatopsis iheyensis sp. nov., isolated from soil on Iheya Island, Japan.

A novel actinomycete, designated strain OK19-0408T, was isolated from soil collected on Iheya island, Okinawa prefecture, Japan. Using the polyphasic taxonomic approach, comparing 16S rRNA gene sequences, the new isolate was found to be most closely related to Amycolatopsis vancoresmycina JCM12675T (98.71 %). Phylogenetic analyses using 16S rRNA sequences indicated that strain OK19-0408T was clustered with Amycolatopsis australiensis JCM15587T. However, digital DNA-DNA hybridization analyses indicated a low relatedness, in the range of 33.9-34.7 %, between strain OK19-0408T and these closely related strains. Strain OK19-0408T contained meso-diaminopimelic acid and whole-cell sugars consisting of arabinose and galactose. The acyl type of the peptidoglycan was acetyl and mycolic acids were absent in strain OK19-0408T. The major menaquinone was MK-9(H4) and hydroxy-phosphatidylethanolamine was detected as the predominant phospholipid. The predominant cellular fatty acid was iso-C16 : 0. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on the polyphasic approach, strain OK19-0408T represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis iheyensis sp. nov. is proposed. The type strain of the type species is OK19-0408T (=NBRC115671T=TBRC16040T).

Mycelial differentiation linked avermectin production in Streptomyces avermitilis studied with Raman imaging.

Streptomyces avermitilis is a gram-positive bacterium that undergoes complex physiological and morphological differentiation during its life cycle, which has implications in secondary metabolite production. Avermectin, produced by S. avermitilis, is widely used as an anthelmintic and insecticidal agent. In this study, we have applied Raman microspectroscopic imaging to elucidate the correlation between production of avermectin and the morphological differentiation in S. avermitilis. We demonstrate distinctive variations in the localization of secondary metabolites at various stages of morphological differentiation. Under solid culture, avermectin was detected in the mycelia formed at the later stages of morphological differentiation (e.g., spore-bearing mycelium and spiral spore chains), but not in the early-stage substrate mycelium. On the contrary, under liquid culture condition, avermectin was found concentrated in the mycelial pellet formed at the early MII stage of differentiation. Furthermore, the chemical profiles of the mycelia were substantially different depending on the culture condition. Raman spectra corresponding to proteins, lipids, and cytochrome were observed in the mycelia irrespective of the stage of morphological differentiation, however, carotenoid was observed under solid culture condition particularly in spore-bearing mycelium and spiral spore chains. KEY POINTS: • Avermectin production is regulated during mycelial differentiation • Liquid and solid culture conditions affects mycelial differentiation • Raman microspectroscopic analysis reveals localization profiles of avermectin.

Streptomyces mimosae sp. nov., an endophytic actinomycete isolated from the root of Mimosa pudica in Thailand.

An endophytic actinomycete, strain 3MP-10T, isolated from the root of Mimosa pudica was taxonomically studied based upon polyphasic approaches. This strain formed spiral spore chains on aerial mycelia. ll-Diaminopimelic acid, glucose and ribose were found in the whole-cell hydrolysates. It belonged to the genus Streptomyces and was closely related to Streptomyces zhaozhouensis DSM 42101T (98.9 %) and Streptomyces sedi JCM 16909T (98.6 %) based on 16S rRNA gene sequence analysis results. The major menaquinones were MK-10(H8), MK-10(H6) and MK-9(H8). The predominant cellular fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The detected phospholipids were diphosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, phosphatidylethanolamine and phosphatidylglycerol. Strain 3MP-10T had a genome size of 7.2 Mb with a genome G+C content of 73.4 mol%. Results of in silico genome-based similarity analysis revealed ANIb values of 84.94 and 84.77 %, ANIm values of 88.01 and 87.92 %, and dDDH values of 29.9 and 29.6 % when compared with S. zhaozhouensis DSM 42101T and S. sedi JCM 16909T, respectively. Based on the polyphasic approach, digital DNA-DNA relatedness and average nucleotide identity, we propose that the novel actinomycete represents a novel species, Streptomyces mimosae, with type strain 3MP-10T (=JCM 33328T=TISTR 2646T).

Micromonospora pelagivivens sp. nov., a new species of the genus Micromonospora isolated from deep-sea sediment in Japan.

A novel marine actinomycete, designated strain KJ-029T, was isolated from a marine sediment sample (water depth of 226 m) in Kagoshima, Japan. 16S rRNA gene sequence analysis revealed that the new isolate was most closely related to Micromonospora craniellae LHW 63014T (99.3 % similarity). Phylogenetic analyses of the genus Micromonospora based on 16S rRNA gene sequences showed that strain KJ-029T was clustered with Micromonospora craniellae LHW 63014T and Micromonospora endophytica 202201T. However, digital DNA-DNA hybridization analyses presented low levels of relatedness in the range of 24.8-32.9 % between strain KJ-029T and the above closely related strains. The novel strain contained meso-diaminopimelic acid and 3-OH-diaminopimelic acid, d-glutamic acid, glycine and d-alanine in the cell-wall peptidoglycan. The acyl type of the peptidoglycan was glycolyl and mycolic acids were absent. The major menaquinone was MK-9(H4). The whole-cell sugars consisted of glucose, mannose, xylose and ribose. Phosphatidylethanolamine was detected as the major phospholipid and corresponded to phospholipid type II. The predominant cellular fatty acid was iso-C16 : 0. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on the present polyphasic study, strain KJ-029T represents a novel species of the genus Micromonospora, for which the name Micromonospora pelagivivens sp. nov. is proposed. The type strain is KJ-029T (=NBRC 113519T=TBRC 9233T).

Detection of Penicillin G Produced by Penicillium chrysogenum with Raman Microspectroscopy and Multivariate Curve Resolution-Alternating Least-Squares Methods.

Raman microspectroscopy is a minimally invasive technique that can identify molecules without labeling. In this study, we demonstrate the detection of penicillin G inside Penicillium chrysogenum KF425 fungal cells. Raman spectra acquired from the fungal cells had highly overlapped spectroscopic signatures and hence were analyzed with multivariate curve resolution by alternating least-squares (MCR-ALS) to extract the spectra of individual molecular constituents. In addition to detecting spatial distribution of multiple constituents such as proteins and lipids inside the fungal body, we could also observe the subcellular localization of penicillin G. This methodology has the potential to be employed in screening the production of bioactive compounds by microorganisms.

Gordonia oryzae sp. nov., isolated from rice plant stems (Oryza sativa L.).

A novel endophytic actinomycete, designated strain RS15-1ST, was isolated from surface-sterilized stems of Oryza sativa L. collected from Sisaket province, Thailand. The colony of strain was strong orange, catalase-positive and oxidase-negative. Growth occurred at a temperature range of 17-37 °C, at pH 4.0-9.0 and in the presence of 0-13 % (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA sequences showed that strain RS15-1ST belonged to the genus Gordonia and was closely related to Gordonia polyisoprenivorans DSM 44302T (98.8 %) and Gordonia rhizosphera DSM 44383T (98.4 %). The major cellular fatty acids were C16 : 0, C18 : 0 10-methyl (tbsa), C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω9c. The menaquinones were MK-9(H2) and MK-8(H2). Strain RS15-1ST contained meso-diaminopimelic acid, arabinose, galactose, mannose and ribose in whole-cell hydrolysates. The polar lipids of the strain were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, an unidentified polar lipid and two unidentified phospholipids. The DNA G+C content was 66.3 mol%. In silico DNA-DNA hybridization of strain RS15-1ST showed 48.3 and 20.5 % relatedness to its closest neighbours, Gordonia polyisoprenivorans DSM 44302T and Gordonia rhizosphera DSM 44383T, respectively. Based on data of genotypic, phenotypic, phylogenetic and chemotaxonomic analysis, strain RS15-1ST represents a novel species of the genus Gordonia, for which the name Gordonia oryzae sp. nov. is proposed. The type strain is RS15-1ST (=TBRC 8485T=NBRC 113446T).

Amycolatopsis suaedae sp. nov., an endophytic actinomycete isolated from Suaeda maritima roots.

An actinomycete strain, designated 8-3EHSuT, was isolated from surface-sterilised roots of Suaeda maritima, collected from Petchburi province, Thailand. Taxonomic position of strain 8-3EHSuT was studied using a polyphasic approach. Phylogenetic determination based on the 16S rRNA gene sequence similarity showed that strain 8-3EHSuT belongs to the genus Amycolatopsis, with the highest sequence similarity to Amycolatopsis jiangsuensis KLBMP 1262T (96.9 %). The colony of strain 8-3EHSuT was yellowish white. Long straight mycelium breaking down into fragments were observed. Growth occurred at temperatures 15-45 °C and at pH 6.0-10.0. The strain contained meso-diaminopimelic acid, arabinose, galactose and mannose in whole-organism hydrolysates. The predominant menaquinone was MK-9(H4). The major fatty acids were C16 : 0, iso-C16 : 0 and iso-C15 : 0. Polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, two unknown amino lipids and an unknown lipid. The G+C content of genomic DNA was 71.8 mol%. A phylogenetic tree based on 16S rRNA sequences showed that the strain 8-3EHSuT formed a distinct evolutionary linage within the genus Amycolatopsis. Based on analysis results of physiological, biochemical and chemotaxonomic data, strain 8-3EHSuT represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis suaedae sp. nov. is proposed. The type strain is 8-3EHSuT (=TBRC 8488T=NBRC 113449T).

Amycolatopsis eburnea sp. nov., an actinomycete associated with arbuscular mycorrhizal fungal spores.

A novel actinomycete, designated strain GLM-1T, was isolated from arbuscular mycorrhizal fungal spores from Funneliformis mosseae RYA08, collected from Aquilaria crassna Pierre ex Lec. rhizosphere soil in Klaeng, Rayong Province, Thailand. Morphological characteristics of this strain included long chains of rod-like cells and squarish elements. The cell-wall composition of this novel isolate contained meso-diaminopimelic acid. The whole-cell diagnostic sugars were arabinose and galactose. The predominant menaquinone was MK-9(H4). The major fatty acids were iso-C16 : 0 and iso-C15 : 0. Only phosphatidylethanolamine was detected as a polar lipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GLM-1T was closely related to Amycolatopsis rhabdoformis SB026T (99.11 %) with a low DNA-DNA hybridization value of 22.6-34.7 %. Genome sequencing revealed a genome size of 10 Mbp. There were obvious distinctions in the average nucleotide identity values between stain GLM-1T and its closely related strains at around 86-93 % (ANIb) and 89-94 % (ANIm). The digital DNA-DNA hybridization values between strain GLM-1T and type strains of phylogenetically related species were 34-55 %. The G+C content of the genomic DNA was 71.8 mol%. Based on these data, strain GLM-1T is considered to represent a novel species of the genus Amycolatopsis, for which the name Amycolatopsiseburnea sp. nov. is proposed. The type strain is GLM-1T (=TBRC 9315T=NBRC 113658T).

Saccharopolyspora rhizosphaerae sp. nov., an actinomycete isolated from rhizosphere soil in Thailand.

A novel actinomycete, designated as strain H219T, was isolated from rhizosphere soil collected under an Elephant ear plant (Colocasiaesculenta) in Bangkok, Thailand. Strain H219T was characterised using a polyphasic approach. Phylogenetic analysis of the 16S rRNA gene sequences revealed that this isolate was most closely related to Saccharopolyspora tripterygii JCM 32123T (97.6 %), Saccharopolyspora dendranthemae NBRC 108675T (97.5 %) and Saccharopolyspora flava NBRC 16345T (97.5 %). However, DNA-DNA hybridization analyses showed a low relatedness in the range of 39-48 % between the novel isolate and the above closely related strains. The cell-wall peptidoglycan of strain H219T contained meso-diaminopimelic acid. The diagnostic whole-cell sugars consisted of arabinose and galactose. The cellular fatty acid profile mainly comprised iso-C16 : 0, anteiso-C17 : 0, iso-C15 : 0, and 10-methyl C17 : 0. The major menaquinone was MK-9(H4). The detected phospholipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylethanolamine-containing hydroxylated fatty acids and an unknown phospholipid. The DNA G+C content was 70.6 mol%. Strain H219T represented chemotaxonomic and morphological characteristics that were consistent with members of the genus Saccharopolyspora. However, strain H219T could be distinguished from closely related strains by several phenotypic properties. Based on the data from the polyphasic studies, we propose that strain H219T is a novel species within the genus Saccharopolyspora, Saccharopolysporarhizosphaerae sp. nov. The type strain is H219T (=TBRC 8564T=NBRC 113388T).

Streptomyces boninensis sp. nov., isolated from soil from a limestone cave in the Ogasawara Islands.

Actinomycete strain K11-0400T was isolated from a soil sample collected in the Ogasawara Islands (also known as the Bonin Islands), Tokyo, Japan. Mature spore chains of strain K11-0400T had more than 20 spores per chain. The strain contained ll-diaminopimelic acid as the diamino acid in whole-cell hydrolysates, and MK-9(H6) and MK-9(H4) were the predominant menaquinones. The polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol, and no diagnostic whole-cell sugar was detected. The G+C content of the genomic DNA was 72 mol%. These morphological and chemical features of strain K11-0400T indicated that it belonged to the genus Streptomyces. Strain K11-0400T showed the highest 16S rRNA gene sequence similarity to Streptomyces naganishii NBRC 12892T (97.58 %). However, the DNA-DNA relatedness value between strain K11-0400T and the related strain was below 70 %. Based on morphological, cultural and physiological characteristics, and DNA-DNA relatedness data, strain K11-0400T should be classified as a new species of the genus Streptomyces, for which the name Streptomyces boninensis sp. nov. is proposed. The type strain of S. boninensis is K11-0400T (=NBRC 113073T, TBRC 7755T).

Actinomadura barringtoniae sp. nov., an endophytic actinomycete isolated from the roots of Barringtonia acutangula (L.) Gaertn.

A novel actinomycete strain, designated GKU 128T, isolated from the roots of an Indian oak tree [Barringtonia acutangula (L.) Gaertn.] at Khao Khitchakut district, Chantaburi province, Thailand, was characterized by using a polyphasic approach. The strain formed a branched substrate and aerial mycelia which differentiated into straight to flexuous chains of smooth-ornamented spores. Analysis of the cell wall revealed the presence of meso-diaminopimelic acid and N-acetylmuramic acid in the peptidoglycan. The whole-cell sugars were glucose, madurose, mannose, rhamnose and ribose. Mycolic acids were absent. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside. The predominant menaquinones were MK-9(H6), MK-9(H8), MK-9(H0) and MK-9(H4). The major fatty acids were C16 : 0, C18 : 1ω9c and 10-methyl C18 : 0 (tuberculostearic acid). The genomic DNA G+C content was 70.5 mol%. Based on 16S rRNA gene sequence analysis, strain GKU 128T was closely related to the type strains of Actinomadura nitritigenes NBRC 15918T (99.2 % sequence similarity) and Actinomadura fibrosa JCM 9371T (98.7 %). The levels of DNA-DNA relatedness between strain GKU 128T and the closely related type species were less than 19 %. On the basis of phenotypic and genotypic characteristics, strain GKU 128T could be distinguished from its closely related type strains and represents a novel species of the genus Actinomadura, for which the name Actinomadura barringtoniae sp. nov. (=TBRC 7225T=NBRC 113074T) is proposed.

Cryptosporangium eucalypti sp. nov., an actinomycete isolated from Eucalyptus camaldulensis roots.

A novel actinomycete, designated strain EURKPP3H10T, was isolated from surface-sterilized roots of Eucalyptus camaldulensis Dehnh., collected from Kamphaengphet Silvicultural Research Station, Kamphaengphet province, Thailand. The taxonomic position of strain EURKPP3H10T was studied using a polyphasic approach. Phylogenetic evaluation based on 16S rRNA gene sequence analysis showed that strain EURKPP3H10T belongs to the genus Cryptosporangium, with the highest sequence similarity to Cryptosporangium cibodasense LIPI11-2-Ac046T (99.2 %). Colonies of strain EURKPP3H10T were orange yellow. Spherical sporangia with motile spores were observed. The strain contained meso-diaminopimelic acid and acofriose, arabinose, galactose, glucose, mannose, xylose and ribose in whole-cell hydrolysates. The predominant menaquinones were MK-9(H8) and MK-9(H6). The major fatty acids were iso-C16 : 0, C17 : 1ω8c, C18 : 1ω9c and C17 : 0. The polar lipids of the strain were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and unknown lipids. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on comparative analysis of physiological, biochemical and chemotaxonomic data, including DNA-DNA hybridization, strain EURKPP3H10T represents a novel species of the genus Cryptosporangium, for which the name Cryptosporangium eucalypti sp. nov. is proposed. The type strain is EURKPP3H10T (=BCC 77605T=NBRC 111482T).

Three novel species of the genus Kibdelosporangium; Kibdelosporangium kanagawaense sp. nov., Kibdelosporangiumrhizosphaerae sp. nov. and Kibdelosporangium rhizovicinum sp. nov.

The taxonomic positions of three actinomycete isolates, K08-0175T, K10-0543T and K12-0791T, which were isolated from plant root and rhizospheric soil samples were subjected to a polyphasic taxonomic study. On the basis of the results of phylogenetic analysis and morphological and chemotaxonomic characteristics, these three strains were classified as representing members of the genus Kibdelosporangium. These strains were observed to produce both long chains of rod-shaped spores and sporangium-like structures with well-defined walls on aerial hyphae. Phylogenetic position, DNA-DNA hybridization and comparison of the phylogenetically closest relatives revealed that these three strains were clearly distinguishable from each other and from their closest phylogenetic relatives. Therefore, three novel species are proposed as Kibdelosporangium kanagawaense sp. nov. [type strain K08-0175T (=NBRC 112388T=TBRC 6786T)], Kibdelosporangiumrhizosphaerae sp. nov. [type strain K10-0543T (=NBRC 112389T=TBRC 6787T)] and Kibdelosporangium rhizovicinum sp. nov. [type strain K12-0791T (=NBRC 112390T=TBRC 6788T)].

Saccharomonospora colocasiae sp. nov., an actinomycete isolated from the rhizosphere of Colocasia esculenta.

A non-Streptomyces actinomycete, designated as strain S265T, was isolated from rhizosphere collected under an elephant ear plant (Colocasia esculenta) in Bangkok, Thailand. The taxonomic position of this strain was determined by a polyphasic approach. Strain S265T formed single globose spores on long, branching, aerial hyphae. It produced abundant aerial mycelium with green colour. The cell wall contained meso-diaminopimelic acid, and diagnostic whole-cell sugars were arabinose and galactose. Phosphatidylethanolamine and diphosphatidylglycerol were detected predominantly as polar lipids, whereas mycolic acids were not found. The major menaquinone was MK-9(H4), and principal cellular fatty acids were C15 : 1 B, iso-C16 : 1 H, anteiso-C15 : 0 and C15 : 0 2-OH. The DNA G+C content was 69 mol%. According to phylogenetic analysis, strain S265T was clustered with Saccharomonospora glauca K62T (98.1 %) and Saccharomonosporaviridis DSM 43017T (97.1 %) despite its 16S rRNA gene sequence showing the highest similarity value to that of Saccharomonosporaazurea NA-128T (98.6 %). DNA-DNA relatedness values between strain S265T and the closely related strains were in the range of 7-50 %, thus strengthening the evidence derived from the polyphasic study that strain S265T represents a novel species within the genus Saccharomonospora, for which the name Saccharomonosporacolocasiae sp. nov. is proposed. The type strain is S265T (=TBRC 7235T=NBRC 112945T).

Dictyobacter aurantiacus gen. nov., sp. nov., a member of the family Ktedonobacteraceae, isolated from soil, and emended description of the genus Thermosporothrix.

A mesophilic, Gram-stain-positive, spore-forming bacterium that formed branched mycelia was isolated from paddy soil in Gunung Salak (Mount Salak), West Java, Indonesia. This strain, designated S-27T, grew at temperatures between 20 and 37 °C; the optimum growth temperature was 25 to 30 °C, and no growth was observed at 15 or 45 °C. The pH range for growth was pH 3.5 to 8.6; the optimum pH was 6.0, and no growth was observed at pH 3.0 or 9.2. Strain S-27T was able to hydrolyse polysaccharides such as starch, cellulose and xylan. The G+C content of the DNA of strain S-27T was 55.7 mol%. The major fatty acids were iso-C17 : 0 and C16 : 1 2-OH, and the major menaquinone was MK-9 (H2). The cell wall of strain S-27T contained d-glutamic acid, glycine, l-alanine, d-alanine, l-ornithine and ß-alanine in a molar ratio of 1.0 : 1.6 : 1.4 : 0.6 : 0.9 : 1.1. The polar lipids consisted of phosphatidylglycerol, phosphatidylinositol and two glycolipids. The major cell-wall sugar was arabinose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S-27T belongs to the order Ktedonobacterales and is most closely related to Ktedonobacter racemifer SOSP1-21T (89.6 % sequence identity). On the basis of its chemotaxonomic and phenotypic features and phylogenetic position, we concluded that strain S-27T represents a novel genus and species, for which we propose the name Dictyobacter aurantiacus gen. nov., sp. nov. The type strain of Dictyobacter aurantiacus is strain S-27T (=NBRC 109595T=InaCC B312T). Emendation of the description of the genus Thermosporothrix is also provided.

Effect of amifostine, a radiation-protecting drug, on oxygen concentration in tissue measured by EPR oximetry and imaging.

Effect of amifostine, a radiation-protecting drug, on muscle tissue partial pressure of oxygen was investigated by electron paramagnetic resonance spectroscopy and imaging. When amifostine was administered intraperitoneally or intravenously to mice, the linewidth of the electron paramagnetic resonance spectra of the lithium octa-n-butoxy-substituted naphthalocyanine implanted in the mouse leg muscle decreased. Electron paramagnetic resonance oximetry using a lithium octa-n-butoxy-substituted naphthalocyanine probe and electron paramagnetic resonance oxygen mapping using a triarylmethyl radical paramagnetic probe was useful to quantify pressure of oxygen in the tissues of living mice. The result of electron paramagnetic resonance oximetric imaging showed that administration of amifostine could decrease pressure of oxygen in the muscle and also tumor tissues. This finding suggests that lowering pressure of oxygen in tissues might contribute in part to the radioprotection of amifostine.

Actinorhabdospora filicis gen. nov., sp. nov., a new member of the family Micromonosporaceae.

The actinomycete strains K12-0408T and K12-0792 were isolated on CM-cellulose agar from rhizosphere soil of a pteridophytic plant collected in Tokyo prefecture, Japan. Their taxonomic positions were determined using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains K12-0408T and K12-0792 were positioned within the family Micromonosporaceae. The strains formed extensively branched aerial and substrate mycelia. Long chains of cylindrical spores with smooth surfaces were formed on aerial hyphae. The cell wall contained meso-diaminopimelic acid, and galactose, glucose, mannose and ribose were detected in whole-cell hydrolysates. The predominant menaquinones were MK-10(H4) and MK-10(H6). The major cellular fatty acids were anteiso-C17 : 0 and anteiso-C17 : 0 2-OH. The DNA G+C contents of strains K12-0408T and K12-0792 were 69.6 and 69.7 mol%, respectively. Based on data from the present polyphasic taxonomic study, strains K12-0408T and K12-0792 represent a novel genus, for which the name Actinorhabdospora gen. nov. is proposed, with strain K12-0408T (=NBRC 111897T=TBRC 5327T) as the type strain of the type species, Actinorhabdosporafilicis sp. nov.

Rhizobium puerariae sp. nov., an endophytic bacterium from the root nodules of the medicinal plant Pueraria candollei var. candollei.

A Gram-stain-negative, rod-shaped, motile bacterium, PC004T, was isolated from root nodules of the Thai medicinal plant Pueraria candollei var. candollei. 16S rRNA gene sequence analysis indicated that the strain is phylogenetically related to species in the genus Rhizobium, showing highest similarity (96.6 %) with Rhizobium mesosinicum HAMBI 3194T. The phylogenetic tree reconstructed based on 16S rRNA gene sequences showed that strain PC004T forms a cluster with Rhizobium petrolearium KCTC 23288T. Based on atpD, gyrB and recA gene sequences, strain PC004T also showed low similarity ( < 90 %) to reference strains. These phylogenetic data indicate that PC004T may represent a novel species. Strain PC004T also exhibited low DNA-DNA relatedness with R. mesosinicum HAMBI 3194T (8.2 %) and R. petrolearium KCTC 23288T (26.3 %). The DNA G+C content of strain PC004T was 64 mol%, which is within the range reported for the genus Rhizobium. The major fatty acid of PC004T was C18 : 1ω7c with minor amounts of C16 : 0, C16 : 0 3-OH, C18 : 0 3-OH, C18 : 1 2-OH, C19 : 0 cyclo ω8c and summed feature 2. The strain was able to grow at pH 12 and with up to 2 % (w/v) NaCl. Strain PC004T did not nodulate five tested legumes and the nifH and nodC genes were not detected by PCR. Based on the physiological, chemotaxonomic and phenotypic data from this study, strain PC004T represents a novel species of the genus Rhizobium, for which the name Rhizobium puerariae sp. nov. is proposed; the type strain is PC004T ( = BCC 73740T = NBRC 110722T).

Nocardiopsissediminis sp. nov., isolated from mangrove sediment.

A filamentous actinomycete, designated strain 1SS5-02T, was isolated from mangrove sediment collected from Ranong province, Thailand. The strain formed aerial and substrate mycelia composed of long, branched hyphae. Aerial mycelia differentiated into non-motile, rod-shaped spores. The organism contained meso-diaminopimelic acid and no diagnostic sugars in whole-cell hydrolysates. The predominant menaquinones were MK-11(H4), MK-11(H6) and MK-11(H8). Polar lipids comprised phosphatidylcholine, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unidentified phospholipids and four unidentified lipids. The major fatty acids were iso-C16 : 0, C18 : 1ω9c, 10-methyl C18 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 73.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 1SS5-02T belonged to the genus Nocardiopsis. The strain showed the highest degree of 16S rRNA gene sequence similarity with 'Nocardiopsis mangrovei' HA11166 (97.9 %) and Nocardiopsis trehalosi VKM Ac-942T (97.8 %). However, strain 1SS5-02T could be distinguished from its nearest phylogenetic relatives in the genus Nocardiopsis on the basis of DNA-DNA relatedness values and the combination of phenotypic properties. On the basis of polyphasic taxonomy, strain 1SS5-02T is considered to represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis sediminis sp. nov. is proposed. The type strain is 1SS5-02T (=BCC 75410T=NBRC 110934T).