<Emphasis Type=Italic>Septoria oleandriicola</Emphasis> sp. nov., a new species from <Emphasis Type=Italic>Nerium oleander</Emphasis> in Turkey

Septoria oleandriicola sp. nov. is described and illustrated from leaves of Nerium oleander, collected in Turkey.     

<Emphasis Type=Italic>Natronincola ferrireducens</Emphasis> sp. nov., and <Emphasis Type=Italic>Natronincola peptidovorans</Emphasis> sp. nov., new anaerobic alkaliphilic peptolytic iron-reducing bacteria isolated from soda lakes

Two novel strains of obligately alkaliphilic (pH 7.5–10.2, optimum pH 8.4–8.8) anaerobic spore-forming rod-shaped bacteria, Z-0511 and Z-7031, were isolated from enrichment cultures obtained from the iron-reducing (Lake Khadyn, Tyva) and cellulolytic (Lake Verkhnee Beloe, Buryatia) bacterial communities, respectively. The organisms ferment peptides and do not ferment proteins and amino acids, with the exception of histidine and glutamate utilized by strain Z-0511. The major fermentation products were acetate and propionate for strain Z-0511 and formate and acetate for strain Z-7031, respectively. Carbohydrates and fermentable organic acids could not serve as substrates, except for pyruvate in the case of strain Z-7031. Nitrogen and sulfur compounds were not utilized as electron acceptors by the strains grown on medium with yeast extract. Strain Z-0511 utilized fumarate, crotonate, and EDTA-Fe(III) as electron acceptors. Anthraquinone-2,6-disulfonate (quinone) and Mn(IV) were utilized by both strains, as well as amorphous ferric hydroxide (AFH), which was reduced to iron sesquioxides and magnetite. The presence of AFH stimulated growth; it enhanced the yield of the fermentation products and changed the quantitative ratios of these products. According to a phylogenetic analysis of the 16S rRNA gene sequences and the phenotypic characteristics of the new strains, they were classified as new species of the genus Natronincola, Natronincola ferrireducens sp. nov. Z-0511^T (= VKM B-2402, = DSM 18346) and Natronincola peptidovorans sp. nov. Z-7031^T (= VKM B-2503, = DSM 18979).     

<Emphasis Type=Italic>Septobasidium parviflorae</Emphasis> sp. nov. on <Emphasis Type=Italic>Pinus parviflora</Emphasis> from Japan

Septobasidium parviflorae sp. nov. on Pinus parviflora is described and illustrated. This species is characterized by its whitish-gray, gray to dark gray-colored fungus body with an indeterminate margin, hyphal strands, and cylindrical basidia with long sterigmata. This is the first report of Septobasidium occurring on a member of the genus Pinus in Japan.     

<Emphasis Type="Italic">Arhodomonas recens</Emphasis> sp. nov., a halophilic alkane-utilizing hydrogen-oxidizing bacterium from the brines of flotation enrichment of potassium minerals

A halophilic nonpigmented rod-shaped (0.8–1.0 × 2.0–2.5 μm), gram-negative bacterium with a single polar flagellum (strain RS91) was isolated from acidic brines of flotation enrichment of potassium minerals (Silvinit Co., Solikamsk, Russia). The strain grew in the media with 2 to 25% NaCl (optimum at 10–12%), 20–45°C (optimum at 37°C), and pH 5.5–8.5 (optimum 6.5–7.5). It was an aerobe or facultative anaerobe incapable of fermentation. The strain was characterized by the absence of growth on glucose, fructose, and citrate, extensive aerobic growth on n-hexadecane and in the mineral medium with H₂ + O₂ + CO₂ in the gas phase, anaerobic nitrate reduction with acetate or hydrogen (under H₂ + CO₂ + N₂), and variable fatty acid composition. The DNA G+C content was 68.2 mol %. Phylogenetic analysis based on 16S rRNA gene sequencing revealed that while strain RS91 was most closely related to Arhodomonas aquaeolei HA-1^T (98.3%) and Nitrococcus mobilis (98.1%), it was only remotely related to the halophilic phototroph Halorhodospira halophila (90.6%). Based on the combination of its phenotypic and genotypic characteristics, the organism was classified as a new species of the genus Arhodomonas, family Ectothiorhodospiraceae with the proposed name Arhodomonas recens sp. nov. The type strain is RS91^T (= IEGM 796^T = VKPM B-11280^T).     

<Emphasis Type=Italic>Stenotrophomonas panacihumi</Emphasis> sp. nov., isolated from soil of a ginseng field

The study isolated a Gram-negative, rod-shaped, non-motile bacterium from the soil of a ginseng field in Daejeon, South Korea and characterized it to determine its taxonomic position. Phylogenetic analysis, based on the 16S rRNA gene sequence, revealed that strain MK06^T belongs to the family Xanthomonadacea, and showed the highest degree of sequence similarity to Stenotrophomonas rhizophila e-p10^T (98.6%), Xanthomonas campestris LMG 568T (98.0%), Stenotrophomonas maltophilia ATCC 1d3637^T (97.3%), and Stenotrophomonas humi R-32729^T (96.9%). Chemotaxonomic data revealed that strain MK06^T possesses ubiquinone Q-8 as the predominant respiratory lipoquinone, which is common in the genus Stenotrophomonas, and that the predominant fatty acids were 15:0 iso (41.1%), 15:0 anteiso (12.6%), and 17:1 iso ω9c (8.6%). The results of physiological and biochemical tests clearly demonstrated that strain MK06^T represents a distinct species and supported its affiliation with the genus Stenotrophomonas. Based on these data, MK06^T (KCTC, 22893^T; JCM, 16536^T; KEMB, 9004-002^T) should be classified as the type strain for a novel species, for which we propose the name Stenotrophomonas panacihumi sp. nov.     

<Emphasis Type=Italic>Spirosoma luteolum</Emphasis> sp. nov. isolated from water

A novel Gram-negative and rod-shaped bacterial strain, designated as 16F6E^T, was isolated from a water sample. Cells were yellowish in color and catalase- and oxidase-positive. The strain grew at 10–37°C (optimum at 25°C) but not at 4 and 42°C, and pH 5–7 (optimum at pH 7). It showed moderate resistance to gamma-ray irradiation. Comparative phylogenetic analysis showed that strain 16F6E^T belonged to the family Cytophagaceae of the class Cytophagia. Furthermore, this isolate showed relatively low 16S rRNA gene sequence similarities (90.7–93.1%) to the members of the genus Spirosoma. The major fatty acids were summed feature 3 (C_16:1 ω7c/C_16:1 ω6c), C_16:1 ω5c, C_16:0 N alcohol, and C_16:0. The polar lipid profile indicated presence of phosphatidylethanolamine, unknown aminophospholipids, an unknown amino lipid, unknown phospholipids, and unknown polar lipids. The predominant isoprenoid quinone was MK-7. The genomic DNA G+C content of strain 16F6E^T was 56.5 mol%. Phenotypic, phylogenetic, and chemotaxonomic properties indicated that isolate 16F6E^T represents a novel species within the genus Spirosoma, for which the name Spirosoma luteolum sp. nov. is proposed. The type strain is 16F6E^T (=KCTC 52199^T =JCM 31411^T).     

<Emphasis Type=Italic>Limnobacter humi</Emphasis> sp. nov., a thiosulfate-oxidizing,heterotrophic bacterium isolated from humus soil,and emended description of the genus <Emphasis Type=Italic>Limnobacter</Emphasis> Spring <Emphasis Type=Italic>et al.</Emphasis> 2001

Three Gram-negative, strictly aerobic, chemolithoheterotrophic bacterial strains, designated UCM-30, UCM-33, and UCM-39^T, were isolated in South Korea. Based on their 16S rRNA gene sequences, the three isolated strains were found to be similar to Limnobacter thiooxidans CS-K2^T (97.41–97.68%), Limnobacter litoralis KP1-19^T (95.55–95.76%), and various genera belonging to the class Betaproteobacteria (90.34–93.34%). DNA-DNA hybridization showed 79.3–83.9% similarity between the genomic DNA of UCM-39^T, UCM-30, and UCM-33, while the sequence similarity between UCM-39^T and L. thiooxidans KACC 13837T or L. litoralis LMG 24869T was 23.7% and 18.6%, respectively. The DNA G+C content of UCM 39T was 59.7 mol%, the major ubiquinone was Q-8, and the optimal oxidation rate was observed at 10 mM thiosulfate. The major fatty acids (≥ 10%) were summed features 3 (C_16:1 ω7c and/or C_16:1 ω6c) and 8 (C_18:1 ω7c and/or C_18:1 ω6c), and C_16:0. The major polar lipids (diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol) were found in all members of genus Limnobacter. Based on phenotypic, physiological, and phylogenetic analyses, the UCM-39T strain was found to be significantly distinct to represent a novel species affiliated to the genus Limnobacter. We propose to name it Limnobacter humi sp. nov. with the type strain UCM-39^T (=KACC 18574^T =NBRC 111650^T).     

<Emphasis Type=Italic>Nocardiopsis terrae</Emphasis> sp. nov., a halophilic actinomycete isolated from saline soil

A Gram-positive, moderately halophilic, facultatively alkaliphilic, catalase- and oxidase-positive, obligately aerobic, filamentous actinomycete strain, designated YIM 90022^T, was isolated from saline soil collected from the Qaidam Basin, north-west China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the new isolate was a member of the genus Nocardiopsis and the sequence similarities between the isolate and the type strains of members of the genus Nocardiopsis were in the range of 95.1–98.7%. Phenotypic and chemotaxonomic properties of this organism also indicated that strain YIM 90022^T was a member of the genus Nocardiopsis. The strain grew well on most of the media tested, producing yellow-white to deep brown substrate mycelium and white aerial mycelium. Light gray to deep brown diffusible pigments were produced. The substrate mycelium was well developed and fragmented with age; the aerial mycelium produced long, straight to flexuous spore chains with non-motile, smooth-surfaced, rod-shaped spores on them. The strain grew in the presence of 1–15% (w/v) total salts (optimum, 3–5%) and at pH 6.0–10.5 (optimum, pH 8.5) and 10–45°C (optimum, 30°C). Whole-cell hydrolysates of strain YIM 90022^T contained meso-diaminopimelic acid and no diagnostic sugars. The predominant menaquinones were MK-10(H₄), MK-9(H₈), MK-10(H₆) and MK-10(H₈). Polar lipids comprised diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylmethylethanolamine. The major cellular fatty acids were iso-C_16:0, anteiso-C_17:0, 10-methyl-C_18:0 and 10-methyl-C_17:0. The DNA G + C content of strain YIM 90022^T was 71.5 mol%. The combination of phylogenetic analysis, DNA–DNA relatedness data, phenotypic characteristics and chemotaxonomic data supported the suggestion that strain YIM 90022^T represents a new species of the genus Nocardiopsis, for which the name Nocardiopsis terrae sp. nov. is proposed. The type strain is YIM 90022^T (=CCTCC AA 208011^T =KCTC 19431^T).     

<Emphasis Type=Italic>Streptomyces nanningensis</Emphasis> sp. nov. a novel Streptomycete from forest soil

A strain YIM 33098^T (= CCTCC AA001027^T = DSM 41831^T) was isolated from a forest soil sample collected from Nanning in Guangxi Province, China, in the course of screening for producers of new drug lead compounds. This strain was identified by using a polyphasic approach. The results showed that it should be assigned to the genus Streptomyces. An almost complete 16S rRNA gene sequence of the strain was determined and compared with those of representative Streptomyces species. Strain YIM 33098^T was clustered in the same subclade with Streptomyces tendae ATCC19812^T and Streptomyces eurythermus ATCC14975^T. Similarities of strain YIM 33098^T with the two strains were 97.35% and 97.42%, respectively. Based on the phenotypic and genotypic evidence, it is therefore proposed that strain YIM 33098^T should be classified in the genus Streptomyces as a new species under the name of Streptomyces nanningensis sp. nov.     

<Emphasis Type=Italic>Hirsutella proturicola</Emphasis> sp. nov. isolated from a proturan, <Emphasis Type=Italic>Baculentulus</Emphasis> densus (Protura,Hexapoda)

A new species of Hirsutella, H. proturicola, isolated from a subterranean proturan (Baculentulus densus; Protura, Hexapoda), is described and illustrated. Hirsutella proturicola is characterized by producing monoblastic phialides of 24–51.5 × 2.5–5 μm with a slightly roughened neck, fusiform and curved conidia of 9–18 × 2.5–4 μm that have a truncate base and a papillate projection often capped with sheath-like mucilage, and pluricellular, globose to subglobose chlamydospores of 21–48 × 21–41.5 μm. This species is morphologically and phylogenetically close to H. rostrata, an acaropathogenic species, but can be distinguished from the size of the phialides and the size and shape of the conidia.     

An endophytic <Emphasis Type=Italic>Neurospora</Emphasis> sp. from <Emphasis Type=Italic>Nothapodytes foetida</Emphasis> producing camptothecin

The medicinal plant, Nothapodytes foetida, contains a number of important alkaloids like camptothecin (an anticancer drug molecule), but its concentration is less to meet the existing demand of this important molecule, in an effort for accessible availability of camptothecin. An endophyte (designated ZP5SE) was isolated from the seed of Nothapodytes foetida and was examined as a potential source of anticancer drug lead compound, i.e., camptothecin, when grown in Sabouraud liquid culture media under shake flask conditions. The presence of an anticancer compound (camptothecin) in this fungus was confirmed by chromatographic and spectroscopic methods in comparison with authentic camptothecin. Isolated endophyte (Neurospora crassa) producing camptothecin may become an easily accessible source for the production of a precursor anticancer drug molecule in the future at a large scale. Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2008, Vol. 44, No. 2, pp. 225–231. The text was submitted by the authors in English.     

<Emphasis Type=Italic>Ceratocystis ficicola</Emphasis> sp. nov., a causal fungus of fig canker in Japan

The causal fungus of Ceratocystis canker of fig in Japan is described as Ceratocystis ficicola sp. nov. This species is characterized by galeated ascospores and is similar to Ceratocystis fimbriata sensu lato. However, the perithecia of the new species are much larger than those of C. fimbriata. The fungus grows more rapidly than C. fimbriata sensu stricto at 25°C but more slowly at 30°C. Molecular analysis of the nucleotide sequences of rDNA ITS regions showed that C. ficicola is phylogenetically placed in the clade of C. fimbriata s. l. but is easily distinguishable from other species of C. fimbriata s. l.     

The presence of <Emphasis Type=Italic>Zygina</Emphasis> sp. and <Emphasis Type=Italic>Puccinia myrsiphylli</Emphasis> reduces survival and influences oviposition of <Emphasis Type=Italic>Crioceris</Emphasis> sp.

A leaf beetle, Crioceris sp. (Coleoptera: Chrysomelidae), was introduced into Australia as a biological control agent of bridal creeper (Asparagus asparagoides L. Druce) during October 2002. Rearing of Crioceris sp. is labour intensive therefore all releases of Crioceris sp. have been under 1000 individuals, which may be too low to ensure establishment if high mortality and high competition with other agents occurs. The aim of this study is to understand how the presence of two well-established biocontrol agents, a rust fungus (Puccinia myrsiphylli (Thuem) Wint [Basidiomycota: Uredinales]) and a leafhopper (Zygina sp. [Hemiptera: Cicadellidae]), might influence Crioceris sp. establishment. Crioceris sp. neonate larvae were placed on bridal creeper plants with or without the leafhopper and/or rust. The number of larvae that pupated was reduced by 38 and 65% in the presence of the rust fungus and leafhopper, respectively and by 45% in the presence of both agents. As the area infected by the rust increased the area damaged by the leafhopper decreased. The rust appeared to be negatively impacted by the presence of the leafhopper. In a second experiment, female Crioceris sp. adults were given a choice between uninfested bridal creeper plants and those infested with the rust or the leafhopper. The females preferred to lay their eggs on plants without leafhoppers but did not seem to be deterred by the presence of the rust. Consequently, the performance and impact of Crioceris sp. on bridal creeper may be reduced if populations overlap with the other biocontrol agents in the field.     

<Emphasis Type=Italic>Ectothiorhodospira magna</Emphasis> sp. nov., a new large alkaliphilic purple sulfur bacterium

Two strains of purple sulfur bacteria of the family Ectothiorhodospiraceae were isolated from moderately saline steppe lakes (with pH above 9.0) of the Transbaikal region (strain B7-7) and Mongolia (strain M10). The cells of the novel strains were spiral-shaped, 2.0–3.2 × 9.6–20.0 μm, motile due to a polar tuft of flagella. Photosynthetic pigments were represented by bacteriochlorophyll a and carotenoids of the spirilloxanthin series. Photosynthetic membranes were represented by long strands of lamellae distributed throughout the whole cell; unlike most Ectothiorhodospiraceae species, the membranes were not packed into regular stacks. Bacteria were capable of weak growth on sulfide and slow grow on hydrogen under photoautotrophic conditions. The best growth was noted on sulfide in the presence of acetate and bicarbonate. Thiosulfate did not stimulate phototrophic growth, even in the presence of organic substrates. The new isolates were alkaliphiles growing at a pH optimum of 9–10. Growth was possible within a salinity range of 0–80 g/l NaCl, with an optimum at 5–15 g/l NaCl. The morphology, the structure of the photosynthetic apparatus (strands of lamellae), and the physiology of the new strains were similar to those of Thiorhodospira sibirica. However, analysis of the 16S rRNA gene sequences demonstrated that the studied isolates were closely related to the type strain Ectothiorhodospira shaposhnikovii (99% similarity) of the family Ectothiorhodospiraceae, whereas the level of similarity between the new strains and Thiorhodospira sibirica was only 94–95%. According to the results of DNA-DNA hybridization, the DNA-DNA homology level between the tested strains was almost 100%; the similarity between the new isolates and the type strain Ectothiorhodospira shaposhnikovii was only 58%. The isolates differed from other representatives of the genus Ectothiorhodospira in the structure of the gene encoding the key enzyme of autotrophic CO₂ fixation, ribulose-1,5-bisphosphate carboxylase (RuBisCo), which was similar to the RuBisCo genes of members of another family of sulfur bacteria, Chromatiaceae. The new isolates of purple bacteria were described as a new species of the genus Ectothiorhodospira, Ect. magna sp. nov. with the type strain B7-7^T (= VKM B-2537 = DSM 22250).     

Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Cryopreservation of <Emphasis Type=Italic>Robinia</Emphasis><Emphasis Type=Italic>pseudoacacia</Emphasis>

Cryopreservation of Robinia pseudoacacia explants by vitrification achieved 78% survival following the stepwise preculture of shoot tips in (0.3 + 0.5 + 0.7 M) sucrose with a 80 min incubation in PVS2; compared to 87% survival after desiccation of explants to 30% water content, following 3 days alginate bead (with glycerol and sucrose treatments) preculture in 0.7 M sucrose.     

<Emphasis Type="Italic">Hoeflea siderophila</Emphasis> sp. nov., a new neutrophilic iron-oxidizing bacterium

A new representative of neutrophilic iron-oxidizing bacteria was isolated from the iron-containing sediments of the brackish low-temperature iron-rich spring of the Staraya Russa Resort (Novgorod region, Russia). The cells of strain Hf1 were thin, slightly curved rods, motile by means of a single polar flagellum. The bacterium reproduced by binary division and was capable of producing rosettes. Optimal growth was observed within the pH range of 6.2–8.5 (with an optimum at 7.5), at 9–38°C (with an optimum at 30°C), and in the salinity range of 0.1–8.5% NaCl (with an optimum at 1%). The organism was a facultative anaerobe. The strain was capable of mixotrophic and organoheterotrophic growth. Fe(II) oxidation occurred under anaerobic conditions via reduction of NO₃⁻ and N₂O, or under microaerobic conditions with oxygen as an electron acceptor. According to phylogenetic analysis based on the comparison of the 16S rRNA gene sequences, the strain was closest to the organotrophic marine bacterium Hoeflea phototrophica (98.5% similarity). The level of DNA-DNA homology with the type species of the genus Hoeflea was 19%. The DNA G + C base content was 57.5 mol %. According to its phenotypic and chemotaxonomic properties, as well as to the results of phylogenetic analysis, strain Hf1 was classified into the genus Hoeflea of the family Phyllobacteriaceae, order Rhizobiales of the phylum Alphaproteobacteria as a novel species, Hoeflea siderophila sp. nov. The type strain is Hf1^T (=DSM 21587 = VKM A7094). The GenBank accession number for the 16S rRNA gene sequences of strain Hf1^T is EU670237.