Effects of maternal vitamin B12 deficiency from end of gestation to weaning on the growth and haematological and immunological parameters in mouse dams and offspring

Vitamin B₁₂-deficiency may induce specific symptoms as neurological alterations and unspecific symptoms such as anaemia and growth retardation. In this study, maternal vitamin B₁₂ deficiency from end of gestation to weaning was evaluated in mouse dams, which was provoked by feeding a vitamin B₁₂-deficient diet. The animals were divided into two groups (control and deficient). The control group received the vitamin B₁₂-deficient diet supplemented with commercial vitamin B₁₂. Compared to the control, the vitamin B₁₂-deficient dams and their offspring showed a significant decrease of body weight (by 20 and 39%, respectively), serum vitamin B₁₂ concentration (by 61 and 67%, respectively), haematological values as haematocrit (25 and 26%, respectively), and IgA producer cells (by 36 and 54%, respectively). In both, vitamin B₁₂-deficient mouse dams and their offspring, histological alterations of small intestine were observed, whereas growth retardation occurred in the offspring only. This experimental murine model allows assessing the incidence of maternal cobalamin deficiency in offspring and would be useful for evaluating novel adjuncts such as functional foods to prevent vitamin B₁₂ deficiency.     

Partial purification of gibberellin oxidases from spinach leaves

Four enzyme activities catalyzing the following oxidative steps in the gibberellin (GA) biosynthetic pathway have been extracted from spinach (Spinacia oleracea L.) leaves after exposure to 8 long days: GA₁₂ → GA₅₃ → GA₄₄ → GA₁₉ → GA₂₀. Two of these, GA₅₃ oxidase and GA₁₉ oxidase, were separable from the other two, GA₄₄ oxidase and GA₁₂ 13-hydroxylase, by anion exchange high performance liquid chromatography (HPLC). Apparent molecular weights of the four enzymes as determined by gel filtration HPLC are: GA₁₂ 13-hydroxylase, 28,400; GA₅₃ oxidase, 42,500; GA₄₄ oxidase, 38,100; GA₁₉ oxidase, 39,500. GA₄₄ oxidase was purified approximately 100-fold in 0.3% yield by a combination of ammonium sulfate fractionation, anion exchange HPLC, phenyl-Sepharose chromatography and gel filtration HPLC.     

Lytic Enzyme from Streptomyces globisporus 1829 Strain

The maximum yield of lytic enzyme was obtained from shake flask cultures of Streptomyces globisporus 1829 which were grown at 30°DC for 48 hr in a medium containing 2% dextrin, 0.5% soybean meal, 0.2% polypeptone, 0.5% Na₂HP0₄. 12H₂0, 0.1% KH₂P0₄, 0.1% MgS0₄·7H₂0, 1.0% NaCI and 0.02% CaCl₂, pH 7.5. The activity of successively transferred substrains of St. globisporus 1829 gradually decreased. However, a mutant strain obtained by ultra-violet irradiation has been shown not to have lost any lytic activity for 2 years. The enzyme exhibited maximum activity at 60°C in the pH range of 6 to 6.5 and was lytic against the intact cells of Streptococci, Lactobacilli and Bacilli but inert against the intact cells of Staphylococcus aureus and Micrococcus lysodeikticus.     

Epicuticular waxes from Agropyron dasystachyum,agropyron riparium and Agropyron elongatum

Epicuticular waxes from whole plants of Agropyron dasystachyum var. psammophylum, A. riparium and A. elongatum contain hydrocarbons (5–8 %), long chain esters (12–15%) and free acids (2–5%). The major esters are C₃₄C₅₆ esters derived from C₁₆C₃₀ acids and alcohols (1-hexacosanol is the major alcohol) but C₃₁, C₃₃ and C₃₅ esters (3–11%) are also present. The latter esters are C₁₈ and C₂₀ acid esters of C₁₃ and C₁₅ 2-alkanols. A. dasystachyum wax contains 2% free alcohols, that of A. riparium contains 17% and that of A. elongatum 11% (1-hexacosanol is the major alcohol in each). Diesters (2%), C₈C₁₂ diols esterified by (E)-2-alkenoic acids, are present in A. riparium wax. Hentriacontane-14,16-dione is present: 29% in A. dasystachyum wax and 32% in A. riparium wax, but only 5% in A. elongatum wax. 25-Oxohentriacontane-14,16-dione forms 14% of A. dasystachyum wax and 27% of A. elongatum wax but the oxo β-diketones of A. riparium wax (5%) consist of both 10-oxo- and 25-oxohentriacontane-14,16-diones in the ratio 4:1. Hydroxy β-diketones of the waxes are 25- and 26-hydroxyhentriacontane-14,16-diones; in A. dasystachyum (20%) the ratio is 3:1, in A. elongatum (20%) the ratio is 9:1 but in A. riparium (5%) it is ca 1:2. The configuration of the hydroxyl group in the 26-hydroxy β-diketone is opposite to that in the 25-hydroxy derivative. The unusual composition of the oxygenated β-diketones of A. riparium confirms that this species should be regarded as separate from A. dasystachyum. Wax from A. elongatum also contains 4-hydroxy-25-oxohentriacontane-14,16-dione (4%) and an unusual oxo-β-ketol, 18-hydroxy-7,16-hentriacontanedione (2%), both these components are probably derived biosynthetically from the 25-oxo β-diketone which is the major component of this wax. Syntheses of racemic 18-hydroxy-7,16-hentriacontanedione and of a model β-ketol, 12-hydroxy-10-pentacosanone, are described.     

Photosynthesis controls of CO2 efflux from maize rhizosphere

The effects of different shading periods of maize plants on rhizosphere respiration and soil organic matter decomposition were investigated by using a ¹³C natural abundance and ¹⁴C pulse labeling simultaneously. ¹³C was a tracer for total C assimilated by maize during the whole growth period, and ¹⁴C was a tracer for recently assimilated C. CO₂ efflux from bare soil was 4 times less than the total CO₂ efflux from planted soil under normal lighting. Comparing to the normal lighting control (12/12 h day/night), eight days with reduced photosynthesis (12/36 h day/night period) and strongly reduced photosynthesis (12/84 h day/night period) resulted in 39% and 68% decrease of the total CO₂ efflux from soil, respectively. The analysis of ¹³C natural abundance showed that root-derived CO₂ efflux accounted for 82%, 68% and 56% of total CO₂ efflux from the planted soil with normal, prolonged and strongly prolonged night periods, respectively. Clear diurnal dynamics of the total CO₂ efflux from soil with normal day-night period as well as its strong reduction by prolonged night period indicated tight coupling with plant photosynthetic activity. The light-on events after prolonged dark periods led to increases of root-derived and therefore of total CO₂ efflux from soil. Any factor affecting photosynthesis, or substrate supply to roots and rhizosphere microorganisms, is an important determinant of root-derived CO₂ efflux, and thereby, total CO₂ efflux from soils. ¹⁴C labeling of plants before the first light treatment did not show any significant differences in the ¹⁴CO₂ respired in the rhizosphere between different dark periods because the assimilate level in the plants was high. Second labeling, conducted after prolonged night phases, showed higher contribution of recently assimilated C (¹⁴C) to the root-derived CO₂ efflux by shaded plants. Results from ¹³C natural abundance showed that the cultivation of maize on Chromic Luvisol decreased soil organic matter (SOM) mineralization compared to unplanted soil (negative priming effect). A more important finding is the observed tight coupling of the negative rhizosphere effect on SOM decomposition with photosynthesis.     

Simulation of fermentation conditions for vitamin B12 biosynthesis from whey

Vitamin B₁₂ production in fermentation of Propionibacterium shermanii and Propionibacterium arl AKU 1251 in whey permeate medium has been studied. The observed results and simulated expected values obtained by fitting statistical equations to the recorded data showed that 24 h old inoculum, 5 mg iron l^?1 and 4% whey lactose were optimal for vitamin B₁₂ biosynthesis in both strains when fermentation was carried out under anerobic (84 h) and aerobic (84 h) conditions at 30°C. The supplementation of whey medium with 0.5% (NH₄)₂HPO₄ enhanced further the metabolite yield; however, the preference for a mixed carbon source (lactose + d-glucose or lactose + d-fructose) at different levels varied in the strains under study. P. shermanii, under optimal cultural conditions, was found to be a better strain than Propionibacterium arl AKU, 1251 in fermenting whey lactose for product (vitamin B₁₂) formation.     

Shewanella litorisediminis sp. nov., a gammaproteobacterium isolated from a tidal flat sediment

A Gram-negative, facultatively anaerobic, non-motile and rod-shaped bacterial strain, designated SMK1-12^T, was isolated from a tidal flat sediment on the western coast of Korea. Phylogenetic analyses based on 16S rRNA and gyrB gene sequences showed that strain SMK1-12^T belonged to the genus Shewanella, clustering with the type strain of Shewanella amazonensis. Strain SMK1-12^T exhibited the highest 16S rRNA gene sequence similarity value (97.0 %) and the highest gyrB sequence similarity value (87.8 %) to S. amazonensis SB2B^T, respectively. Strain SMK1-12^T contained simultaneously both menaquinones and ubiquinones; the predominant menaquinone was MK-7 and the predominant ubiquinones were Q-7 and Q-8. The major fatty acids (>10 % of the total fatty acids) detected in strain SMK1-12^T were the MIDI system summed feature 3 (iso-C_15:0 2-OH and/or C_16:1 ω7c), iso-C_15:0, C_17:1 ω8c and C_16:0. The DNA G+C content of strain SMK1-12^T was 58.0 mol% and its mean DNA–DNA relatedness value with S. amazonensis ATCC 700329^T was 15 ± 4.6 %. Differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, revealed that strain SMK1-12^T is distinguishable from recognized Shewanella species. On the basis of the data presented, strain SMK1-12^T is considered to represent a novel Shewanella species, for which the name Shewanella litorisediminis sp. nov. is proposed. The type strain is SMK1-12^T (=KCTC 23961^T = CCUG 62411^T).     

Pigmentiphaga soli sp. nov., a bacterium isolated from soil

Strain BS12^T, a Gram-negative motile bacterium, was isolated from soil in South Korea and characterized to determine its taxonomic position. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the strain belonged to the family Alcaligenaceae in the class Betaproteobacteria. The highest degree of sequence similarities of strain BS12^T were found with Pigmentiphaga litoralis JSM 061001^T (98.3%), Pigmentiphaga daeguensis K110^T (98.2%), and Pigmentiphaga kullae K24^T (98.1%). Chemotaxonomic data revealed that strain BS12^T possessed ubiquinone-8, which is common in the family Alcaligenaceae, and the predominant fatty acids were C_16:0, C_17:0 cyclo, summed feature 3 (C_16:1 ω6c/ω7c), and summed feature 8 (C_18:1 ω6c/ω7c). The major polar lipids of strain BS12^T were phosphatidylethanolamine and phosphatidylglycerol. Based on these data, BS12^T (=KCTC 23577^T =JCM 17666^T =KEMB 9004-082^T) should be classified as a type strain of a novel species, for which the name Pigmentiphaga soli sp. nov. is proposed.     

Jeotgalibacillus aurantiacus sp. nov., a novel orange-pigmented species with a carotenoid biosynthetic gene cluster,isolated from wetland soil

A Gram-stain-positive, orange-pigmented, rod-shaped and flagellated bacterial strain T12^T was isolated from wetland soil in Kunyu Mountain Wetland in Yantai, China. The strain was able to grow at 15–40 °C (optimum 37 °C), at 0.0–9.0% NaCl (optimum 2%, w/v) and at pH 5.5–9.0 (optimum 8.5). A phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain T12^T is a member of the family Planococcaceae, sharing 97.6% and 97.1% sequence similarity with the type strains of Jeotgalibacillus salarius and Jeotgalibacillus marinus, respectively. Genome-based analyses revealed a genome size of 3,506,682 bp and a DNA G?+?C content of 43.7%. Besides, the genome sequence led to 55.0–74.6% average amino acid identity values and 67.8–74.7% average nucleotide identity values between strain T12^T and the current closest relatives. Digital DNA-DNA hybridization of strain T12^T with the type strains of Jeotgalibacillus proteolyticus and J. marinus demonstrated 19.0% and 20.3% relatedness, respectively. The chemotaxonomic analysis showed that the sole quinone was MK-7. The predominant cellular fatty acids were iso-C_15:0, anteiso-C_15:0, C_16:1ω7c alcohol and iso-C_14:0. The polar lipids consisted of an unidentified aminolipid, phosphatidylglycerol, diphosphatidylglycerol and two unidentified phospholipids. Based on the polyphasic characterization, strain T12^T is considered to represent a novel species, for which the name Jeotgalibacillus aurantiacus sp. nov. is proposed. The type strain is T12^T (=?KCTC 43296 ^T?=?MCCC 1K07171^T).

     

Binding,Activation, and Solubilization of the Ca2+-ATPase from Sarcoplasmic Reticulum by Nonionic Detergents

Interactions between delipidated Ca²⁺-ATPase from sarcoplasmic reticulum and four nonionic detergents—dodecyl octaoxyethyleneglycol monoether (C₁₂E₈), Triton X-100, Brij 58, and Brij 35—were characterized with respect to activation of ATPase activity, binding, and solubilization. C₁₂E₈ and Triton X-100 activated the delipidated ATPase to at least 80% of the original activity at the critical micelle concentrations (CMCs), whereas Brij 58 and Brij 35 activated no more than 10% of the original activity. The inability of Brij 58 and Brij 35 to activate the delipidated enzyme was probably a result of reduced binding of these detergents below the CMCs; both detergents exhibited a sixteenfold reduction in binding at the CMC compared with C₁₂E₈. The two Brij detergents were also unable to solubilize the delipidated enzyme and form monomers, as determined by sedimentation experiments. Thus the reduced binding levels of these detergents may result from an inability to overcome protein/protein interactions in the delipidated preparation. However, the Brij detergents were capable of solubilizing active enzyme from membrane vesicles, although with lower efficiency than C₁₂E₈ and Triton X-100. These results suggest that Brij 58 and 35 may be useful for solubilization of membrane proteins without disrupting protein/protein interactions, while Triton X-100 and C₁₂E₈ are more useful when bulk solubilization is the goal.     

Pedobacter seoulensis sp. nov., isolated from soil of a bamboo field

A Gram-stain negative, strictly aerobic, motile by gliding, rod-shaped and yellow pigmented strain THG-G12^T was isolated from soil of a bamboo field in Seoul, Republic of Korea. Strain THG-G12^T was observed to grow well at 20–28 °C and pH 7.0–7.5 in the absence of NaCl on nutrient agar. Based on 16S rRNA gene sequence comparisons, strain THG-G12^T was found to be most closely related to Pedobacter ginsengisoli Gsoil 104^T (97.5 % sequence similarity), Pedobacter steynii WB2.3-45^T (97.4 %), Pedobacter metabolipauper WB2.3-71^T (97.2 %), Pedobacter nyackensis NWG-II14^T (97.2 %), Pedobacter caeni LMG 22862^T (97.1 %) and Pedobacter duraquae WB2.1-25^T (97.0 %), but DNA relatedness between strain THG-G12^T and its phylogenetically closest neighbours was below 9.5 %. The G+C content of the genomic DNA was determined to be 39.9 mol%. The only isoprenoid quinone detected in strain THG-G12^T was menaquinone-7 (MK-7). The major component in the polyamine pattern was sym-homospermidine. The major polar lipids were found to be phosphatidylethanolamine, unidentified phosphoglycolipids, unidentified aminophosphoglycolipids, unidentified aminolipids and unidentified lipids. Strain THG-G12^T showed the presence of two ceramide phosphorylethanolamines (CerPE-2′ and CerPE-2″), dihydrosphingosines and an unidentified ceramide as the major ceramide. The major fatty acids were identified as summed feature 3 (as defined by the MIDI system; C_16:1 ω7c and/or C_16:1 ω6c) and iso-C_15:0. These data support the affiliation of strain THG-G12^T to the genus Pedobacter. The results of physiological and biochemical tests enabled strain THG-G12^T to be differentiated genotypically and phenotypically from the recognized species of the genus Pedobacter. Therefore, the novel isolate represents a novel species, for which the name Pedobacter seoulensis sp. nov. is proposed, with THG-G12^T as the type strain (=KACC 17529^T =JCM 19363^T).     

Eubacterium aggreganssp. nov., a New Homoacetogenic Bacterium from Olive Mill Wastewater Treatment Digestor

A strictly anaerobic, homoacetogenic, Gram-positive, non spore-forming bacterium, designated strain SR12^T(T=type strain), was isolated from an anaerobic methanogenic digestor fed with olive mill wastewater. Yeast extract was required for growth but could also be used as sole carbon and energy source. Strain SR12^Tutilized a few carbohydrates (glucose, fructose and sucrose), organic compounds (lactate, crotonate, formate and betaine), alcohols (methanol), the methoxyl group of some methoxylated aromatic compounds, and H₂+CO₂. The end-products of carbohydrate fermentation were acetate, formate, butyrate, H₂and CO₂. End-products from lactate and methoxylated aromatic compounds were acetate and butyrate. Strain SR12^Twas non-motile, formed aggregates, had a G+C content of 55 mol % and grew optimally at 35°C and pH 7.2 on a medium containing glucose. Phylogenetically, strain SR12^Twas related toEubacterium barkeri, E. callanderi, andE. limosumwithE. barkerias the closest relative (similarity of 98%) with which it bears little phenotypic similarity or DNA homology (60%). On the basis of its phenotypic, genotypic, and phylogenetic characteristics, we propose to designate strain SR12^TasEubacterium aggreganssp. nov. The type strain is SR12^T(=DSM 12183).     

The biosynthesis of 12α-hydroxylated gibberellins in a cell-free system from cucurbita maxima endosperm

A previously unknown pathway for the biosynthesis of 12α-hydroxylated gibberellins was found in a cell-free system from Cucurbita maxima endosperm. The microsome fraction converts the gibberellin precursor GA_12-aldehyde simultaneously to GA₁₂ and 12α-hydroxy-GA₁₂-aldehyde. The ratio of these products is pH-dependent: above pH 6.5, the production of GA₁₂ is favoured, whilst below pH 6.5, 12α-hydroxy-GA₁₂-aldehyde is the predominant product. 12α-Hydroxy-GA₁₂-aldehyde is converted further by soluble enzymes to 12α-hydroxy-GA₁₄, 12α-hydroxy-GA₁₅, 12α-hydroxy-GA₃₇ and several unidentified products. This conversion is optimal between pH 6.0 and 6.5 in contrast to the previously known conversion of GA₁₂-aldehyde to GA₄₃ by soluble enzymes, which is optimal at pH 7.5. GA₅₈, a major 12α-hydroxylated endogenous constituent of C. maxima endosperm, was not obtained when 12α-hydroxy-GA₁₂-aldehyde was used as a substrate, but it was obtained together with GA₄ when GA₉ was incubated with a preparation containing both microsomal and soluble enzymes.     

Description and genomic characterization of Nocardioides bruguierae sp. nov., isolated from Bruguiera gymnorhiza

Strains BSK12Z-3^T and BSK12Z-4, two Gram-stain-positive, aerobic, non-spore-forming strains, were isolated from Shankou Mangrove Nature Reserve, Guangxi Zhuang Autonomous Region, China. The diagnostic diamino acid in the cell-wall peptidoglycan of strain BSK12Z-3^T was LL-diaminopimelic acid and MK-8(H₄) was the predominant menaquinone. The polar lipids comprised diphosphatidylglycerol (DPG), phosphatidylglycerol (PG) and phospholipid (PL). The major fatty acids was iso-C_16:0. Phylogenetic analysis based on 16S rRNA gene sequences suggested that the two strains fell within the genus Nocardioides, appearing most closely related to Nocardioides ginkgobilobae KCTC 39594^T (97.5–97.6 % sequence similarity) and Nocardioides marinus DSM 18248^T (97.4–97.6 %). Genome-based phylogenetic analysis confirmed that strains BSK12Z-3^T and BSK12Z-4 formed a distinct phylogenetic cluster within the genus Nocardioides. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of strains BSK12Z-3^T, BSK12Z-4 with their most related species N. marinus DSM18248^T were within the ranges of 77.2–77.3 % and 21.3–21.4 %, respectively, clearly indicated that strains BSK12Z-3^T, BSK12Z-4 represented novel species. Strains BSK12Z-3^T and BSK12Z-4 exhibited 99.9 % 16S rRNA gene sequence similarity. The ANI and dDDH values between the two strains were 97.8 % and 81.1 %, respectively, suggesting that they belong to the same species. However, DNA fingerprinting discriminated that they were not from one clonal origin. Based on phylogenomic and phylogenetic analyses coupled with phenotypic and chemotaxonomic characterizatons, strains BSK12Z-3^T and BSK12Z-4 could be classified as a novel species of the genus Nocardioides, for which the name Nocardioides bruguierae sp. nov., is proposed. The type strain is BSK12Z-3^T (=CGMCC 4.7709^T = JCM 34554^T).     

亲和层析纯化肌质网Ca2+－ATP酶

建立了一种亲和层析纯化肌质网Ca²⁺-ATP酶的方法．用非离子型去污剂C₁₂E₈ 溶解肌质网,再通过反应红－120琼脂糖亲和层析柱使肌质网Ca²⁺-ATP酶纯度从粗品中的65％提高到99％,并具有较高ATP水解活性．经SDS－聚丙烯酰胺凝胶电泳检测,为电泳纯．     

Identification of a novel calcium binding protein from bovine brain

A novel Ca²⁺ binding protein, named caligulin, was extracted from the heat-treated 100 000 × g supernatant of bovine brain and purified to electrophoretic homogeneity. The apparent M_r of caligulin determined on sodium dodecyl sulfate polyacrylamide gels was 24 000. Analysis by gel filtration chromatography indicated an apparent M_r of 33 000, suggesting a monomeric protein. Amino acid composition data demonstrated the presence of 25% acidic residues, 12% basic residues and 10% leucine. In the presence of 1 mM MgCl₂ and 0.15 M KCl, caligulin bound 1 mol Ca²⁺/mol protein with half-maximal binding at about 0.2 μM Ca²⁺.     

Gibberellin biosynthesis in cell-free extracts from developing Cucurbita maxima embryos and the identification of new endogenous gibberellins

Gibberellin (GA) biosynthetic pathways from GA₁₂-aldehyde, GA₁₂ and GA₅₃ were investigated in cell-free systems from developing embryos of Cucurbita maxima L. Gibberellin A₁₂-aldehyde and GA₁₂ were converted to GA₂₅, putative 12α-hydroxyGA₂₅, GA₁₃ and GA₃₉ as main products. Minor products were GA₄, GA₃₄ and, when GA₁₂ was the substrate, putative 12α-hydroxyGA₁₂. The intermediates GA₁₅ and GA₂₄ accumulated at low protein concentrations. The influence of various factors on GA₁₂ metabolism was examined. At low 2-oxoglutarate and ascorbate concentrations, or at acid pH, 3β-hydroxylated products predominated, whereas with increasing 2-oxoglutarate and ascorbate concentrations, or at neutral pH, the yield of 12α-hydroxylated GAs increased. Gibberellin A₅₃ was metabolised mainly to the C₂₀-GAs GA₄₄, GA₁₉, GA₁₇, GA₂₃ and GA₂₈, with the C₁₉-GAs GA₂₀, GA₁ and GA₈ as minor products. Only C₁₉-GAs were 2β-hydroxylated, which is a main characteristic of the embryo systems. In addition to GA₁₃, GA₂₅, GA₃₉, GA₄₃, GA₄₉, GA₅₈, GA₇₄, 12α-hydroxyGA₂₅ and GA₃₉ 3-isovalerate, which were known previously from embryos of C. maxima, GA₁, GA₄, GA₁₇, GA₂₈, GA₃₇, GA₃₈, GA₄₈, GA₈₅, 12α-hydroxyGA₃₇ and putative 12α-hydroxyGA₄₃ were identified as endogenous components by full-scan capillary gas chromatography-mass spectrometry and Kovats retention indices. Evidence for putative 2β-hydroxyGA₂₈ and GA₂₃ was also obtained but it was less conclusive because of contamination.     

<Emphasis Type="Italic">Vibrio injenensis</Emphasis> sp. nov., isolated from human clinical specimens

Vibrio species are well known as motile, mostly oxidase-positive, facultative anaerobic Gram-negative bacteria. They are abundant in aquatic environments and are a common cause of human infections including diarrhea, soft tissue diseases, and bacteremia. Here, two Gram-negative bacteria, designated M12-1144^T and M12-1181, were isolated from human clinical specimens and identified using a polyphasic taxonomic approach. Phylogenetic study based on 16S rRNA gene sequence analysis revealed that the isolates belong to the genus Vibrio, and are closely related to Vibrio metschnikovii KCTC 32284^T (98.3%) and Vibrio cincinnatiensis KCTC 2733^T (97.8%). The major fatty acids were summed feature 3 (C_16:1 ω7c/C_16:1 ω6c, 38.0%), C_16:0 (23.0%), and summed feature 8 (C_18:1 ω7c or C_18:1 ω6c, 19.3%) and major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The G + C content of the genomic DNA was determined to be 44.1 mol%. DNA–DNA relatedness between the two newly isolated strains and V. metschnikovii KCTC 32284^T and V. cincinnatiensis KCTC 2733^T was between 42.6 to 47.5%. The similarities of genome-to-genome distance between M12-1144^T and related species ranged from 18.4-54.8%. Based on these results, a new species of the genus Vibrio, Vibrio injenensis is proposed. The type strain is M12-1144 ^T(=KCTC 32233^T =JCM 30011^T).     

Nitrite reduction and carbohydrate metabolism in plastids purified from roots of Pisum sativum L.

Nitrate reduction in roots and shoots and exchange of reduced N between organs were quantitatively estimated in intact 13-d-old seedlings of two-row barley (Hordeum vulgare L. cv. Daisengold) using the ¹⁵N-incorporation model (A. Gojon et al. (1986) Plant Physiol. 82, 254–260), except that NH ₊ ⁴ was replaced by NO _- ² . N-depleted seedlings were exposed to media containing both nitrate (1.8 mM) and nitrite (0.2 mM) under a light-dark cycle of 12:12 h at 20°C; the media contained different amounts of ¹⁵N labeling. Experiments were started either immediately after the beginning (expt. 1) or immediately prior to the end (expt. 2) of the light period, and plants were sampled subsequently at each light-dark transition throughout 36 h. The plants effectively utilized ¹⁵NO _- ³ and accumulated it as reduced ¹⁵N, predominantly in the shoots. Accumulation of reduced ¹⁵N in both experiments was nearly the same at the end of the experiment but the accumulation pattern in roots and shoots during each 12-h period differed greatly depending on time and the light conditions. In expt. 1, the roots accounted for 31% (light), 58% (dark), and 9% (light) of nitrate reduction by the whole plants, while in expt. 2 the contributions of the root were 82% (dark), 20% (light), and 29% (dark), during each of the three 12-h periods. Xylem transport of nitrate drastically decreased in the dark, but that of reduced N rather increased. The downward translocation of reduced ¹⁵N increased while nitrate reduction in the root decreased, whereas upward translocation decreased while nitrate reduction in the shoot increased. We conclude that the cycling of reduced N through the plant is important for N feeding of each organ, and that the transport system of reduced N by way of xylem and phloem, as well as nitrate reduction by root and shoot, can be modulated in response to the relative magnitude of reduced-N demands by the root and shoot, with the one or the other predominating under different circumstances.Symbols Anl accumulation of reduced ¹⁵N from ¹⁵NO _- ³ in ¹⁴NO _- ³ -fed roots of divided root system - Ar accumulation in root of reduced ¹⁵N from ¹⁵NO _- ³ - As accumulation in shoot of reduced ¹⁵N from ¹⁵NO _- ³ - Rr ¹⁵NO _- ³ reduction in root - Rs ¹⁵NO _- ³ reduction in shoot - Tp translocation to root of shoot-reduced ¹⁵N from ¹⁵NO _- ³ in phloem - Tx translocation to shoot of root-reduced ¹⁵N from ¹⁵NO _- ³ in xylem