<Emphasis Type="Italic">Sphingomonas rosea</Emphasis> sp. nov. and <Emphasis Type="Italic">Sphingomonas swuensis</Emphasis> sp. nov., rosy colored <Emphasis Type="Italic">β</Emphasis>-glucosidase-producing bacteria isolated from soil

Two strains PB196^T and PB62^T of Gram-negative, non-motile, and non-spore-forming bacteria, were isolated from soil in South Korea and characterized to determine their taxonomic positions. 16S rRNA gene sequence analysis showed that the two strains belonged to the genus Sphingomonas. The highest degree of sequence similarity of strain PB196^T was found with PB62^T (98.9%), Sphingomonas humi PB323^T (98.9%), Sphingomonas kaistensis PB56^T (98.2%), and Sphingomonas astaxanthinifaciens TDMA-17^T (98.0%). The highest degree of sequence similarity of strain PB62^T was found with Sphingomonas humi PB323^T (98.8%), Sphingomonas astaxanthinifaciens TDMA-17^T (98.2%), and Sphingomonas kaistensis PB56^T (98.1%). Chemotaxonomic data revealed that they possessed ubiquinone-10 (Q-10) as common in the genus Sphingomonas, that the predominant fatty acids were summed feature 7 (C_18:1 ω7c/ω9t/ω12t), summed feature 4 (C_16:1 ω7c/C_15:0 iso 2OH), C_16:0, and C_17:1 ω6c, and that they contained sphingoglycolipid, phosphatidylglycerol (PG), and phosphatidyle-thanolamine (PE) in common but they showed difference for diphosphatidylglycerol (DPG). Based on these data, PB196^T (=KCTC 12339^T =JCM 16604^T) and PB62^T (=KCTC 12336^T =JCM 16605^T =KEMB 9004-005^T) should be classified as type strains of two novel species, for which the names Sphingomonas rosea sp. nov. and Sphingomonas swuensis sp. nov. are proposed, respectively.     

<Emphasis Type="Italic">Luteoamylascus aculeatus</Emphasis> (Pezizomycetes,Pezizaceae): a new genus and species near the <Emphasis Type="Italic">Pachyphlodes</Emphasis>–<Emphasis Type="Italic">Amylascus</Emphasis> lineage

The new taxon Luteoamylascus aculeatus described in this article is proposed to accommodate two collections of a hypogeous ascomycete from central Spain, characterized by a tomentose yellowish peridium, labyrinth-like gleba filled with whitish hyphae, and intensely reacting amyloid asci. ITS, 28S, and RPB2 data suggest that this new taxon is an independent lineage proposed here as the new genus Luteoamylascus. Until now, this lineage was only known from ectomycorrhizal root tips and mitotic spore mats. In phylogenetic analyses, the Luteoamylascus lineage is placed close to the genera Amylascus, Pachyphlodes, and Scabropezia. Morphological data suggest an affinity with Amylascus.     

Two new ascomycetous anamorphic yeast species related to <Emphasis Type="Italic">Candida friedrichii</Emphasis>—<Emphasis Type="Italic">Candida jaroonii</Emphasis> sp. nov., and <Emphasis Type="Italic">Candida songkhlaensis</Emphasis> sp. nov.—isolated in Thailand

In a study of yeast diversity in Thailand, eight strains of hitherto undescribed anamorphic yeasts were isolated: four from insect frass, two from Marasmius sp. fruiting bodies, one from a flower, and one from jackfruit exudates. Phylogenetic analysis of the D1/D2 domain of 26S ribosomal DNA nucleotide sequences indicated that the eight strains represented two new species related to Candida friedrichii. Genetic separation of the two new species was further supported by DNA-DNA hybridization analysis, which resulted in between-species similarity values of less than 48%, and by electrophoretic karyotyping. The two new species are C. jaroonii sp. nov. (type strain, ST-300(T) = NBRC 103209(T) = BCC 11783(T) = CBS 10790(T)) and C. songkhlaensis sp. nov. (type strain, ST-328(T) = NBRC 103214(T) = BCC 11804(T) = CBS 10791(T)).     

A review of the species of the genus <Emphasis Type="Italic">Adoxomyia</Emphasis> Kertész, 1907 (Diptera,Stratiomyidae) with bicolorous antennae and a description of a new species, <Emphasis Type="Italic">Adoxomyia subruficornis</Emphasis> sp. n. from Middle Asia

Morphological features of the species of the genus Adoxomyia Kertész, 1907 with bicolorous antennae in females were studied for the first time. Significant diagnostic characters are discussed, e.g., the structure of the head, the type of pubescence on the frons and face, the shape and coloration of the scutellar spines, and the morphology of the genitalia (the synstemum, the basal part of the epandrium, and the aedeagus). A new species, Adoxomyia subruficornis sp. n., is described from Uzbekistan and Tajikistan; it differs from A. ruficornis (Loew, 1873) in a dense pale pubescence on the frons, pale scutellar spines, elongate median projection of the synstemum, and rounded epandrial processes.     

<Emphasis Type="Italic">Acinetobacter kyonggiensis</Emphasis> sp. nov., a <Emphasis Type="Italic">β</Emphasis>-glucosidase-producing bacterium,isolated from sewage treatment plant

A Gram-negative, non-motile bacterium, designated KSL5401-037^T, was isolated from a sewage treatment plant in Gwangju in the Republic of Korea and was characterized using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis showed that strain KSL5401-037^T belonged to the genus Acinetobacter in the family Moraxellaceae of the Gammaproteobacteria (Brisou and Prevot, 1954). According to a 16S rRNA gene sequence analysis, it was closely related to Acinetobacter johnsonii ATCC 17909^T (97.3%), A. bouvetii 4B02^T (97.2%), and A. beijerinckii 58a^T (96.8%). Chemotaxonomic data revealed that strain KSL5401-037^T possesses an ubiquinone system with Q-8 as the predominant compound and C_16:0 (19.2%), C_18:1 ω9c (19.5%), and summed feature 3 (C_16:1 ω6c / C_16:1 ω7c, 34.1%) as the predominant cellular fatty acids. The major polar lipids detected in strain KSL5401-037^T were diphosphatidylglycerol (DPG) and, phosphatidylethanolamine (PE), followed by phosphatidylglycerol (PG) and moderate amounts of phosphatidylcholine and phosphatidylserine. The G+C content of the genomic DNA was 41.2–42.1 mol%. Strain KSL5401-037^T exhibited relatively low levels of DNA-DNA relatedness with respect to A. johnsonii DSM 6963^T (17.7%) and A. bouvetii 4B02^T (9.3%). The DNA-DNA relatedness values, biochemical, and physiological characteristics of strain KSL5401-037^T strongly support its genotypic and phenotypic differentiation from other recognized type strains of the genus Acinetobacter. Based on these data, strain KSL5401-037^T (JCM 17071^T =KEMC 5401-037^T) should be classified in the genus Acinetobacter as a type strain of novel species, for which the name Acinetobacter kyonggiensis sp. nov. is proposed.     

<Emphasis Type="Italic">Condyloderes kurilensis</Emphasis> sp. nov. (Kinorhyncha: Cyclorhagida)—a new deep water species from the abyssal plain near the Kuril-Kamchatka Trench

A new species of centroderid kinorhynchs, Condyloderes kurilensis sp. nov., collected from muddy sediment at the abyssal plain near the Kuril-Kamchatka Trench, North-West Pacific, is described and illustrated using light and electron microscopy. Previously, members of the genus Condyloderes were found only in nearly shallow water enviroments—C. multispinosus (North Sea), C. paradoxus (Bengal Bay), C. setoensis (Tanabe Bay, Japan), C. storchi (Argentina coast), and C. megastigma (Korea Strait). The new species is characterized by the presence of lateroventral cuspidate spines only on trunk segment 8 in both sexes, paired ventromedial appendages (tubules) on trunk segments 7 and 8 in females only, and dorsolateral spines on trunk segment 10 in males only. Condyloderes kurilensis sp. nov. constitutes the sixth species of the genus Condyloderes described so far and the only second abyssal species of the Kinorhyncha described from the North-West Pacific. A key to the six species of the genus Condyloderes is proposed.     

Synthesis of disaccharides using β-glucosidases from <Emphasis Type="Italic">Aspergillus niger</Emphasis>, <Emphasis Type="Italic">A. awamori</Emphasis> and <Emphasis Type="Italic">Prunus dulcis</Emphasis>

Objective

Glucose conversion into disaccharides was performed with β-glucosidases from Prunus dulcis (β-Pd), Aspergillus niger (β-An) and A. awamori (β-Aa), in reactions containing initial glucose of 700 and 900 g l^?1.

Results

The reactions’ time courses were followed regarding glucose and product concentrations. In all cases, there was a predominant formation of gentiobiose over cellobiose and also of oligosaccharides with a higher molecular mass. For reactions containing 700 g glucose l^?1, the final substrate conversions were 33, 38, and 23.5% for β-An, β-Aa, and β-Pd, respectively. The use of β-An yielded 103 g gentiobiose l^?1 (15.5% yield), which is the highest reported for a fungal β-glucosidase. The increase in glucose concentration to 900 g l^?1 resulted in a significant increase in disaccharide synthesis by β-Pd, reaching 128 g gentiobiose l^?1 (15% yield), while for β-An and β-Aa, there was a shift toward the synthesis of higher oligosaccharides.

Conclusion

β-Pd and the fungal β-An and β-Aa β-glucosidases present quite dissimilar kinetics and selective properties regarding the synthesis of disaccharides; while β-Pd showed the highest productivity for gentiobiose synthesis, β-An presented the highest specificity.

     

Retiolitid graptolites from the collection of Hermann Jaeger II: <Emphasis Type="Italic">Cometograptus</Emphasis>, <Emphasis Type="Italic">Spinograptus</Emphasis> and <Emphasis Type="Italic">Plectograptus</Emphasis>

Graptolites from the Jaeger collection at the Museum für Naturkunde (Berlin, Germany) provide important information on structural details of Silurian (Wenlock–Ludlow) retiolitids as well as for the biostratigraphic and biogeographic distribution of these magnificent graptolites. Species of the genera Cometograptus, Spinograptus and Plectograptus are described from isolated glacial boulder material, collected in northern Germany and from shale specimens found in the Lower Graptolite Shale of Thuringia. The biostratigraphic placement of material derived from glacial erratic boulders, however, is far from being precise. The fauna associated with the neotype of Plectograptus macilentus in the ‘Unterer Graptolithenschiefer’ of Thuringia is discussed and illustrated. Cometograptus alfeisenacki from the Cyrtograptus lundgreni Biozone is recognized as a new species. The genus is discovered for the first time in North German glacial erratic boulders.     

Sequence Analysis of the <Emphasis Type="Italic">Lactoferrin</Emphasis> Gene and Variation of <Emphasis Type="Italic">g</Emphasis>.<Emphasis Type="Italic">7605C</Emphasis>→<Emphasis Type="Italic">T</Emphasis> in 10 Chinese Indigenous Goat Breeds

Much attention has been focused on the study of lactoferrin at the protein or nucleotide level in mice, humans, and cattle, but little is known about it in goats. The goat LF gene from 5' UTR to exon 17 was amplified, and the variation of g.7605C→T in 10 Chinese indigenous goat breeds was analyzed. Among the three ruminant species (cattle, sheep, and goats), the intron-exon distribution pattern was similar, and all the exons had the same length, but the length of introns varied greatly due to insertions or deletions. The frequency of allele T at g.7605C→T (50.12%) was a little higher than that of allele C (49.88%), and the genotype distribution differed greatly between goat populations. The g.7605C→T site showed higher genetic diversity in goat populations. The genetic differentiation was 0.0783, and gene flow was 2.9433 among the 10 Chinese indigenous goat populations.     

<Emphasis Type="Italic">Ceratonykus oculatus</Emphasis> gen. et sp. nov., a new dinosaur (?Theropoda,Alvarezsauria) from the Late Cretaceous of Mongolia

A new dinosaur, Ceratonykus oculatus gen. et sp. nov. (Parvicursoridae, Alvarezsauria), from the Upper Cretaceous (Baruungoyot Formation) of Mongolia is described based on a fragmentary skeleton. It differs from other alvarezsaurians in many characters, including the short femora, long tarsometatarsals, and considerably reduced third metatarsals. The carpometacarpals contain spikelike bones. The natural endocast of the new taxon shows large acoustic tubercles, the ventral position of the optic lobes in the midbrain, and the absence of a vertical flexure at the brain floor. The data obtained cast doubt on the taxonomic position of alvarezsaurians in the Theropoda.     

Characteristics of β-lactamases and their genes (<Emphasis Type="Italic">blaA</Emphasis> and <Emphasis Type="Italic">blaB</Emphasis>) in <Emphasis Type="Italic">Yersinia intermedia</Emphasis> and <Emphasis Type="Italic">Y. frederiksenii</Emphasis>

Background  

The presence of β-lactamases in Y. enterocolitica has been reported to vary with serovars, biovars and geographical origin of the isolates. An understanding of the β-lactamases in other related species is important for an overall perception of antibiotic resistance in yersiniae. The objective of this work was to study the characteristics of β-lactamases and their genes in strains of Y. intermedia and Y. frederiksenii, isolated from clinical and non-clinical sources in India.     

Cryopreservation of cell suspension cultures of <Emphasis Type="Italic">Taxus</Emphasis> × <Emphasis Type="Italic">media</Emphasis> and <Emphasis Type="Italic">Taxus floridana</Emphasis>

Different lines of cell suspension cultures of Taxus × media Rehd. and Taxus floridana Nutt. were cryopreserved with a two-step freezing method using a simple and inexpensive freezing container instead of a programmable freezer. Four to seven days old suspension cell cultures were precultured in growth medium supplemented with 0.5 M mannitol for 2 d. The medium was then replaced with cryoprotectant solution (1 M sucrose, 0.5 M glycerol and 0.5 M dimethylsulfoxide) and the cells incubated on ice for 1 h. Before being plunged into liquid nitrogen, cells were frozen with a cooling rate of approximately −1 °C per min to −80 °C. The highest post-thaw cell viability was 90 %. The recovery was line dependent. The cryopreservation procedure did not alter the nuclear DNA content of the cell lines. The results indicate that cryopreservation of Taxus cell suspension cultures using inexpensive freezing container is possible.     

Conjugal transfer using the bacteriophage ϕC31 <Emphasis Type="Italic">att</Emphasis>/<Emphasis Type="Italic">int</Emphasis> system and properties of the <Emphasis Type="Italic">attB</Emphasis> site in <Emphasis Type="Italic">Streptomyces ambofaciens</Emphasis>

To facilitate molecular genetic studies of Streptomyces ambofaciens that produces spiramycin, a commercially important macrolide antibiotic used in human medicine against Gram-positive pathogenic bacteria, the conditions for the conjugal transfer of DNA from E. coli to S. ambofaciens were established using a bacteriophage ϕC31 att/int system. The transconjugation efficiency of S. ambofaciens varied with the medium used; the highest frequency was obtained on AS-1 medium containing 10 mM MgCl₂ without heat treatment of the spores. In addition, by cloning and sequencing the attB site, we identified that S. ambofaciens contains a single attB site within an ORF coding for a pirin homolog, and its attB site sequence shows 100% nt identity to the sequence of S. coelicolor and S. lividans, which have the highest efficiency in transconjugation using the ϕC31 att/int system.     

<Emphasis Type="Italic">Paracoccus pueri</Emphasis> sp. nov., isolated from Pu’er tea

A Gram-stain negative, aerobic, short rod-shaped, motile by flagella bacterial strain (THG-N2.35^T), was isolated from Pu’er tea. Growth occurred at 10–40 °C (optimum 28 °C), at pH 4–7 (optimum 7) and at 0–5% NaCl (optimum 1%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-N2.35^T were identified as Paracoccus hibisci KACC 18632^T (99.0%), Paracoccus tibetensis CGMCC 1.8925^T (98.7%), Paracoccus beibuensis CGMCC 1.7295^T (98.2%), Paracoccus aestuarii KCTC 22049^T (98.2%), Paracoccus rhizosphaerae LMG 26205^T (98.1%), Paracoccus zeaxanthinifaciens ATCC 21588^T (97.1%), Paracoccus marcusii DSM 11574^T (97.0%). Levels of similarity between strain THG-N2.35^T and other Paracoccus species were lower than 97.0%. DNA-DNA hybridization values between strain THG-N2.35^T and P. hibisci KACC 18632^T, P. tibetensis CGMCC 1.8925^T, P. beibuensis CGMCC 1.7295^T, P. aestuarii KCTC 22049^T, P. rhizosphaerae LMG 26205^T, P. zeaxanthinifaciens ATCC 21588^T, P.marcusii DSM 11574^T were 47.5% (42.3%, reciprocal analysis), 36.1% (32.3%), 24.7% (22.1%), 19.2% (16.3%), 11.3% (8.8%), 11.1% (10.8%), 6.1% (5.8%), respectively. The DNA G+C content of strain THG-N2.35^T was 62.3 mol%. The polar lipids were diphosphatidylglycerol, phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. The quinone was ubiquinone-10 (Q-10). The major fatty acids were C_10:0 3OH, C_16:0, C_18:0 and C_18:1 ω7?. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-N2.35^T represent a novel species of the genus Paracoccus, for which the name Paracoccus pueri sp. nov. is proposed. The type strain is THG-N2.35^T (= KACC 18934^T = CCTCC AB 2016177^T).     

<Emphasis Type="Italic">Adelobotrys atlantica</Emphasis> (<Emphasis Type="Italic">Melastomataceae: Merianieae</Emphasis>): the first species of <Emphasis Type="Italic">Adelobotrys</Emphasis> from Brazil’s Mata Atlântica

Summary  A new species of Adelobotrys DC., A. atlantica Schulman, is described and illustrated. This is the first record of the genus in the Atlantic rainforests of Brazil. The only known specimen of the new species was collected in 1943 close to the sizeable city of Ilhéus in an area that has probably now been converted to managed land. It is therefore possible that A. atlantica is already extinct.     

Biochemical characterization of a raw starch degrading <Emphasis Type="Italic">α</Emphasis>-amylase from the Indonesian marine bacterium <Emphasis Type="Italic">Bacillus</Emphasis> sp. ALSHL3

An Indonesian marine bacterial isolate, which belongs to genus of Bacillus sp. based on 16S rDNA analysis and was identified as Bacillus filicolonicus according to its morphology and physiology, produced a raw starch degrading α-amylase. The partially purified α-amylase using a maize starch affinity method exhibited an optimum pH and temperature of 6.0 and 60°C, respectively. The enzyme retained 72% of its activity in the presence of 1.5 M NaCl. Scanning electron micrographs showed that the α-amylase was capable of degrading starch granules of rice and maize. This α-amylase from Bacillus sp. ALSHL3 was classified as a saccharifying enzyme since its major final degradation product was glucose, maltose, and maltotriose.     

Regeneration of transformed verbena (<Emphasis Type="Italic">Verbena </Emphasis>× <Emphasis Type="Italic">hybrida</Emphasis>) by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

Verbena (Verbena x hybrida), an important floricultural species, was successfully regenerated from stem segments on Murashige and Skoog's basal medium supplemented with thidiazuron and indole-3-acetic acid. A transformation system was developed using cvs. Temari Scarlet, Temari Sakura, Tapien Rose and TP-P2. Agrobacterium tumefaciens strain Agl0 harboring the sGFP gene was infected into stem segments. Transformation efficiency was improved by evaluating and manipulating the age of the plant material, the concentration of kanamycin in the medium during selection, and the length of the culture period in the dark. After 2-3 months of culture on the selection medium, GFP-positive shoots were obtained in all four of the cultivars tested. These shoots were successfully acclimated and set flowers within 2-3 months in a greenhouse. GFP was expressed in all of the organs including the floral parts. Stable genomic transformation was confirmed by Southern blot analysis. No morphological differences were observed between the transformed plants and their host plants.     

Cloning,expression, and characterization of <Emphasis Type="Italic">β</Emphasis>-glucosidase from <Emphasis Type="Italic">Exiguobacterium</Emphasis> sp. DAU5 and transglycosylation activity

A gram-negative bacterium, designated strain DAU5, was isolated from shrimp shell samples because it demonstrated high β-glucosidase activity. Through 16S rDNA gene sequence analysis the strain was identified as belonging to the genus Exiguobacterium. The β-glucosidase gene of Exiguobacterium sp. DAU5 was successfully cloned by the shotgun method. Nucleotide sequence determination by sodium dodecyl sulfate-ployacrylamide gel electrophoresis indicated that the gene for the enzyme contained 1,350 bp, was coded by 450 amino acids, and was 52 kDa. The polypeptide exhibits significant homology with other bacterial β-glucosidases and belongs to the Glycoside Hydrolase Family 1. The β-glucosidase was purified by a His-fusion purification system. The optimal pH and temperature of the enzyme were 7.0 and 45°C, respectively. The enzyme activity was strongly inhibited by Ca²⁺, and Li⁺, K⁺, Zn²⁺, Mg²⁺, Na²⁺, Ni²⁺, and EDTA partially inhibited the enzyme activity. The BglA showed the highest activity with p-NPG and MUG. However, strain DAU5 β-glucosidase, which is for degradation of oligosaccharides, is expected to be useful for the fermentation of cellulose degradation and the transglycosylation of saccharides.     

Mating system and seed variation of <Emphasis Type="Italic">Acacia</Emphasis> hybrid (<Emphasis Type="Italic">A. mangium</Emphasis> × <Emphasis Type="Italic">A. auriculiformis</Emphasis>)

The mating system and seed variation of Acacia hybrid (A. mangium × A. auriculiformis) were studied using allozymes and random amplified polymorphic DNA (RAPD) markers, respectively. Multi-locus outcrossing rate estimations indicated that the hybrid was predominantly outcrossed (mean±s.e. t _m = 0.86±0.01). Seed variation was investigated using 35 polymorphic RAPD fragments. An analysis of molecular variance (AMOVA) revealed the highest genetic variation among seeds within a pod (66%–70%), followed by among pods within inflorescence (29%–37%), and the least variation among inflorescences within tree (<1%). In addition, two to four RAPD profiles could be detected among seeds within pod. Therefore, the results suggest that a maximum of four seeds per pod could be sampled for the establishment of a mapping population for further studies.     

Isolation and functional characterization of <Emphasis Type="Italic">Lycopene β-cyclase</Emphasis> (<Emphasis Type="Italic">CYC-B</Emphasis>) promoter from <Emphasis Type="Italic">Solanum habrochaites</Emphasis>

Background  

Carotenoids are a group of C40 isoprenoid molecules that play diverse biological and ecological roles in plants. Tomato is an important vegetable in human diet and provides the vitamin A precursor β-carotene. Genes encoding enzymes involved in carotenoid biosynthetic pathway have been cloned. However, regulation of genes involved in carotenoid biosynthetic pathway and accumulation of specific carotenoid in chromoplasts are not well understood. One of the approaches to understand regulation of carotenoid metabolism is to characterize the promoters of genes encoding proteins involved in carotenoid metabolism. Lycopene β-cyclase is one of the crucial enzymes in carotenoid biosynthesis pathway in plants. Its activity is required for synthesis of both α-and β-carotenes that are further converted into other carotenoids such as lutein, zeaxanthin, etc. This study describes the isolation and characterization of chromoplast-specific Lycopene β-cyclase (CYC-B) promoter from a green fruited S. habrochaites genotype EC520061.