Environmental Sciences

The MspI restriction-modification system, which recognizes the sequence 5'-CCGG-3', has been previously cloned and sequenced (1). We subcloned the methyltransferase gene (M.MspI) downstream of the ptac promoter in the multicopy vector pUC119 and overexpressed it in E. coli. Upon induction with IPTG, M.MspI constitutes more than 10% of cellular protein. A scheme has been devised to purify large amounts of biologically active M.MspI to apparent homogeneity from these overexpressing E. coli cells. Approximately 0.8 mg of pure M.MspI per gram of cells (wet weight) can be obtained. The apparent molecular weight of M.MspI is 49 kD, by SDS gel electrophoresis and 48-54 kD by gel filtration. At low concentrations (less than 0.4 mg/ml), the methyltransferase is a monomer in solution but at higher concentrations (greater than 3.0 mg/ml) it exists predominantly as a dimer. Polyclonal antibodies raised against M.MspI cross-react with the DNA-methyltransferases of several other restricti...

Biochar addition to agricultural soil has been suggested to mitigate climate change through increased biogenic carbon storage and reduction of greenhouse gas emissions. We measured the fluxes of N2O, CO2, and CH4 after adding 9 t ha−1 biochar on an agricultural soil in Southern Finland in May 2009. We conducted these measurements twice a week for 1.5 months, between sowing and canopy closure, to capture the period of highest N2O emissions, where the potential for mitigation would also be highest. Biochar addition increased CH4 uptake (96% increase in the average cumulative CH4 uptake), but no statistically significant differences were observed in the CO2 and N2O emissions between the biochar amended and control plots. Added biochar increased soil water holding capacity by 11%. Further studies are needed to clarify whether this may help balance fluctuations in water availability to plants in the future climate with more frequent drought periods.▶ Biochar addition to boreal agricultural soil increases CH4 uptake. ▶ Biochar addition to boreal agricultural soil increases water holding capacity. ▶ Biochar addition to boreal agricultural soil had no effect on N2O or CO2 fluxes.

An extensive programme of experiments was conducted in the former USSR on transfer of radionuclides to a wide range of different agricultural animals. Only a few of these studies were made available in the English language literature or taken into account in international reviews of gastrointestinal uptake. The paper gives extended information on Russian research on radionuclide absorption in the gut of farm animals performed in controlled field and laboratory experiments from the 1960s to the current time. The data presented in the paper, together with English language values, will be used to provide recommended values of absorption specifically for farm animals within the revision of the IAEA Handbook of Parameter Values IAEA [International Atomic Energy Agency, 1994. Handbook of Parameter Values for the Prediction of Radionuclide Transfer in Temperate Environments, IAEA technical reports series No. 364. International Atomic Energy Agency, Vienna].

Mutations in human mitochondrial DNA (mtDNA) can cause mitochondrial disease and have been associated with neurodegenerative disorders, cancer, diabetes and aging. Yet our progress toward delineating the precise contributions of mtDNA mutations to these conditions is impeded by the limited availability of faithful transmitochondrial animal models. Here, we report a method for the isolation of mutations in mouse mtDNA and its implementation for the generation of a collection of over 150 cell lines suitable for the production of transmitochondrial mice. This method is based on the limited mutagenesis of mtDNA by proofreading-deficient DNA-polymerase γ followed by segregation of the resulting highly heteroplasmic mtDNA population by means of intracellular cloning. Among generated cell lines, we identify nine which carry mutations affecting the same amino acid or nucleotide positions as in human disease, including a mutation in the ND4 gene responsible for 70% of Leber Hereditary Optic ...

Carcharhinus sorrah (Mülle and Henle, 1839) is a coastal pelagic shark of the tropical and subtropical Indo–West Pacific, reaching 1.6 m total length. The species is widely harvested in line, net, and trawl fisheries over tropical continental shelves. We investigated mtDNA genetic differentiation in C. sorrah across the majority of the species' range, and examined the importance of six major historical and contemporary biogeographic features in shaping population genetic structure in this species. The present study includes dense sampling for a shark species across the Indo–West Pacific, with 349 specimens sampled from 21 collection locations from the northwestern Indian Ocean, Southeast Asia, New Caledonia, and to southerly distribution limits in Australia. Based on 469 base pairs of the control region, we found extensive genetic population structure, with allopatric lineages confined to Australia, New Caledonia, and the remaining surveyed area. Significant genetic subdivisions...

Aquatic macroinvertebrates living in anastomosing lowland rivers use different habitats and respond differently to the hydrological regime. In this paper, the structure and composition of benthic, drifting and marginal macroinvertebrate assemblages are analyzed in the lowland river Ctalamochita (Córdoba, Argentina). The assemblages were studied in an annual cycle; a comparison among the composition of benthos, drift and marginal fauna was carried out; and size structure of the assemblages was characterized. Samples were obtained from two sites: a rural and an urban site. In total 73 taxa of aquatic macroinvertebrates were collected. Benthos was characterized by Chironomidae and Oligochaeta; marginal fauna was mainly constituted by Coleoptera, Heteroptera, Decapoda, the Trichoptera Nectopsyche sp., Ephemeroptera and Odonata. Drifting assemblage was composed by macroinvertebrates from local and remote upstream benthos, and from the marginal zone. Marginal fauna diversity was higher than benthos and drift. Total biomass of the assemblages pooled together was relatively equitably among size classes. Larger size classes consisted of organisms from the marginal zone whereas the smallest ones were composed by benthic and drifting organisms. In the study area there is habitat partitioning in the lateral dimension of the river. Marginal fauna was more diverse due to the asymmetry of transport and deposit processes, which generate a heterogeneous habitat in the bankside. The relation between fine substrate and high current velocity determines an unstable habitat in the central channel, which makes colonization by benthic macroinvertebrates difficult.