Biogeochemical and hydrological controls on carbon export from a forested catchment in central Japan

Here we review research on the links between hydrological processes and the biogeochemical environment controlling the dynamics of dissolved organic carbon (DOC) and dissolved inorganic carbon (DIC) in temperate forested catchments. In addition, we present the results of original experiments. The spatial and temporal changes in DIC and DOC concentrations were investigated in tandem with observations of elementary belowground hydrological processes for a forested headwater catchment in central Japan. The soil CO₂ gas concentration, which is the source of DIC, increased with depth. The hydrological characteristics of groundwater also affected the spatial variation of partial pressure of dissolved CO₂ (pCO₂) in groundwater. The temporal variations in the soil CO₂ gas concentration and the pCO₂ values of groundwater suggested that the dynamics of DIC were strongly affected by biological activity. However, the geographical differences in DIC leaching were affected not only by the link between climatological conditions and biological activity, but also by other factors such as geomorphologic conditions. The DOC concentrations decreased with selective removal of hydrophobic acid during vertical infiltration. The major DOC-removal mechanisms were retention of metal-organic complexes to soil solids in the upper mineral soil layer and decomposition of DOC in the lower mineral soil layer. The responses of the DIC and DOC concentrations to changes in discharge during storm events were explained by the spatial variation in the DIC and DOC concentrations. Seasonal variation, which represents a long-term change, in stream water DOC concentrations was affected not only by the temporal variation in DOC concentrations in the topsoil, which may be affected by biological activity, but also by water movement, which transports DOC from the topsoil to stream water. These results indicate that both a biogeochemical approach and a method for evaluating the hydrological effects on carbon dynamics are critical for clarifying the carbon accumulation-and-release processes in forested ecosystems.     

Identification procedure for Pasteurella pneumotropica in microbiologic monitoring of laboratory animals.

Discrepancies have been recognized in the identification of Pasteurella pneumotropica between testing laboratories. To determine the causes of the differences and to propose a reliable identification procedure for P. pneumotropica, a working group was organized and 69 isolates identified or suspected as P. pneumotropica were collected from 8 laboratories in Japan. These isolates were examined by colony morphology, Gram-staining, the slide agglutination test using two antisera (ATCC35149 and MaR), two commercially available biochemical test kits (ID test, API20NE) and two primer sets of PCR tests (Wang PCR, CIEA PCR). The 69 isolates and two reference strains were divided into 10 groups by test results. No single procedure for P. pneumotropica identification was found. Among tested isolates, large differences were not observed by colony morphology and Gram-straining except for colony colors that depended on their biotypes. Sixty-eight out of 69 isolates were positive by the slide agglutination test using two antisera except for one isolate that tested with one antiserum. The ID test identified 61 out of 69 isolates as P. pneumotropica and there was no large difference from the results of CIEA PCR. From these results, we recommend the combination of colony observation, Gram-straining, the slide agglutination tests with two antisera and biochemical test using the ID test for practical and reliable identification of this organism.     

Multidrug resistance-associated protein 9 (ABCC12) is present in mouse and boar sperm

In order to search for novel components of lipid membrane microdomains involved in neural signalling pathways, mAbs (monoclonal antibodies) were raised against the detergent-insoluble membrane fraction of PC12 (pheochromocytoma) cells. Among the 22 hybrid clones, mAb PR#1 specifically detected a fucoganglioside Fuc(Gal)-GM1 [a-fucosyl(a-galactosyl)-GM1], a ganglioside homologous with GM1a (II3NeuAc,GgOse4Cer), as a novel member of microdomain components with biological functions. In the presence of mAb PR#1 in the culture medium, the outgrowth of neurites was induced in PC12 cells in a dose-dependent manner, with no effects on cell proliferation, suggesting that Fuc(Gal)-GM1 is preferentially involved in PC12 cell neuritogenesis. Effects through Fuc(Gal)-GM1 were different from those through GM1a during differentiation, e.g. under PR#1 treatment on Fuc(Gal)-GM1, round cell bodies with thinner cell processes were induced, whereas treatment with CTB (cholera toxin B subunit), a specific probe for GM1a, produced flattened cell bodies with thicker pro-cesses. Molecular analysis demonstrated that the PR#1-Fuc(Gal)-GM1 pathway was associated with Fyn and Yes of the Src family of kinases, although Src itself was not involved. No association was found with TrkA (tropomyosin receptor kinase A) and ERKs (extracellular-signal-regulated kinases), which are responsible for GM1a-induced differentiation. From these findings, it is suggested that a fucoganglioside Fuc(Gal)-GM1 provides a functional platform distinct from that of GM1a for signal transduction in PC12 cell differentiation.     

Structure and function of H+-ATPase

(1) Extensive studies on proton-translocating ATPase (H⁺-ATPase) revealed that H⁺-ATPase is an energy transforming device universally distributed in membranes of almost all kinds of cells. (2) Crystallization of the catalytic portion (F₁) of H⁺-ATPase showed that F₁ is a hexagonal molecule with a central hole. The diameter of F₁ is about 90 Å and its molecular weight is about 380,000. (3) Use of thermophilic F₁ permits the complete reconstitution of F₁ from its five subunits (, , , , and ) and demonstration of the gate function of the -complex, the catalytic function of (supported by and ), and the H⁺-translocating functions of all five subunits. (4) Studies using purified thermostable F₀ showed that F₀ is an H⁺-channel portion of H⁺-ATPase. The direct measurement of H⁺-flux through F₀, sequencing of DCCD-binding protein, and isolation of F₁-binding protein are described. (5) The subunit stoichiometry of F₁ may be ₃₃. (6) Reconstitution of stable H⁺-ATPase-liposomes revealed that ATP is directly synthesized by the flow of H⁺ driven by an electrochemical potential gradient and that H⁺ is translocated by ATP hydrolysis. This rules out functions for all the hypothetical components that do not belong to H⁺-ATPase in H⁺-driven ATP synthesis. The roles of conformation change and other phenomena in ATP synthesis are also discussed.     

Plant species effect on the spatial patterns of soil properties in the Mu-us desert ecosystem,Inner Mongolia,China

Although Artemisia ordosica Krasch. and Sabina vulgaris Ant. are the dominant shrub species in the Mu-us desert ecosystem, they differ in their botanical traits. We investigated the spatial patterns of soil properties using geostatistical analysis to examine the effect of plant species on these spatial patterns. Comparison among three microsite types (under A. ordosica, under S. vulgaris, and the opening between vegetation) showed that A. ordosica generally had less effect than S. vulgaris on local soil properties. The long life-span, prostrate life-form, and evergreen leaf-habit of S. vulgaris may lead to a higher accumulation of organic and fine materials under S. vulgaris. The range of spatial autocorrelation found in the mass of organic matter on the soil surface was smaller than that of the coverage of S. vulgaris (11.5 m) which corresponded to the canopy patch size of this species, and was longer than the canopy patch size of A. ordosica. The ranges of total C and N, and pH (11.7–15.6 m) were similar to that of S. vulgaris. The range of available P (106.3 m) was comparable to that of the coverage of A. ordosica (86.2 m) considered to be the scale of the distribution of this species. The ranges of silt+clay and exchangeable K, Ca, and Mg (31.0–46.7 m) were not related to plant presence, and were similar to that of topography (43.1 m).     

Importance of hydrophobic interaction between a SoxB-type cytochrome c oxidase with its natural substrate cytochrome c-551 and its mutants

Cytochrome c-551, the electron donor of SoxB-type cytochrome c oxidase in thermophilic bacilli, can be over-expressed in Bacillus thermodenitrificans cells by tranformation with pSTEc551. Several mutant cytochromes c-551 were prepared by site-directed mutagenesis to this expression plasmid. Among them, several Lys residues were changed to Ala/Ser, and we found that these mutant cytochromes retained their activity as substrates, although their K(m) values were 0.04-0.12 microM, depending on the site replaced. In contrast, the C19A mutant cytochrome, which was produced in Brevibacillus choshinensis as a secretion protein, lost its activity as a substrate, suggesting that the fatty acyl-glyceryl residue covalently bound to the cysteine residue of the wild-type c-551 plays a very important role in the activity. The importance of the hydrophobic fatty acid residue for the binding of cytochrome c-551 to the oxidase was also shown by the loss of substrate activity in deacylated cytochrome c-551. These results show the importance of the hydrophobic interaction between this cytochrome and SoxB-type oxidase, despite the fact that the importance of an electrostatic interaction between cytochrome c and mitochondrial cytochrome aa(3) oxidase has already been established.     

Higher diversity and abundance of denitrifying microorganisms in environments than considered previously

Denitrification is an important process in the global nitrogen cycle. The genes encoding NirK and NirS (nirK and nirS), which catalyze the reduction of nitrite to nitric oxide, have been used as marker genes to study the ecological behavior of denitrifiers in environments. However, conventional polymerase chain reaction (PCR) primers can only detect a limited range of the phylogenetically diverse nirK and nirS. Thus, we developed new PCR primers covering the diverse nirK and nirS. Clone library and qPCR analysis using the primers showed that nirK and nirS in terrestrial environments are more phylogenetically diverse and 2–6 times more abundant than those revealed with the conventional primers. RNA- and culture-based analyses using a cropland soil also suggested that microorganisms with previously unconsidered nirK or nirS are responsible for denitrification in the soil. PCR techniques still have a greater capacity for the deep analysis of target genes than PCR-independent methods including metagenome analysis, although efforts are needed to minimize the PCR biases. The methodology and the insights obtained here should allow us to achieve a more precise understanding of the ecological behavior of denitrifiers and facilitate more precise estimate of denitrification in environments.     

Comparison of oxygen consumption in drosophilid flies from different climates

Oxygen consumption at rest was studied in drosophilid species from cool‐temperate, warm‐temperate and subtropical regions to assess whether adaptations to different climates are associated with changes in metabolic rates. In experiments at 23°C using 8‐day‐old males of 28 species, body mass was revealed to be a significant predictor of oxygen consumption. No significant relation was detected between mass‐adjusted oxygen consumption and latitudinal distribution or thermal tolerance by either conventional regression analysis or a phylogenetically based method. The effect of temperature on oxygen consumption was studied with experiments at 15, 18, 23 and 28°C using 8‐ and 24‐day‐old males of four species of each of the montium species subgroup and the subgenus Drosophila. In these experiments, it was confirmed that temperature was a significant predictor of mass‐adjusted oxygen consumption. In both lineages, mass‐adjusted oxygen consumption was not higher in cool‐temperate species than in subtropical species. Thus, adaptations to colder climates are not associated with elevation of metabolic rates in these drosophilid species. The results of the present study also indicate that oxygen consumption is not related to the capacity to walk quickly.