DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Length of Incubation for the Estimation of the Most Probable Number of Nitrifying Bacteria in Soil

Abstract

The Zn content in 8 soil chemical fractions was determined for 21 greenhouse soils and for 8 reference (open field) soils from Kochi Prefecture, Japan to investigate the forms of spontaneously accumulated Zn in the greenhouse soils associated with heavy application of fertilizers and manures. Sequential extraction method was applied to every soil and each Zn fraction was designated as exchangeable (Ex-Zn), Pb-displaceable (Pb-Zn), acid soluble (Aci-Zn), Mn oxide-occluded (MnO-Zn), organically bound (OM-Zn), amorphous Fe oxide-occluded (AFeO-Zn), crystalline Fe oxide-occluded (CFeO-Zn), and residual (Res-Zn) fractions. The Zn content of the greenhouse soils was significantly higher than that of the reference soils in every fraction, except for the CFeO-Zn, and Res-Zn fractions. The Pb-Zn, Aci-Zn, and MnO-Zn fractions showed a difference of more than 60% in the total Zn content between the two soil groups. The amounts of Zn extracted in the Pb-Zn, Aci-Zn, MnO-Zn, AFeO-Zn, and CFeO-Zn fractions of the greenhouse soils increased con-comitantly with the accumulation of applied macro-nutrients. These results indicated that the accumulation of Zn in greenhouse soils caused by intensive fertilization had proceeded through specific adsorption onto or occlusion by the oxides and hydroxides of Fe and Mn in soils.     

Influence of phosphate sorption on dispersion of a Ferralsol

Abstract

Soil dispersion induces soil erosion and colloidal leaching. Nutrients are lost at the same time and this causes water contamination. Phosphate is an essential element for living organisms. Because phosphate influences soil dispersion and it is an important limited resource, this influence must be evaluated well in order to diminish negative effects on soil structure. In this paper, we firstly evaluated the influence of phosphate sorption on soil dispersion by calculating repulsive potential energy between soil particles. Ferralsol, which is a typical soil in rainy tropical regions, was used as the material. The dispersion-flocculation phenomena were investigated with absorbance of soil suspension under different pH, phosphate adsorption and electrolyte concentration in an Na-NO₃-PO₄ system. The repulsive potential energy was calculated based on the diffuse double layer theory and the measured zeta potential. We indicated that the measured absorbance increased with the increase of the repulsive potential energy. The repulsive potential energy increased with increasing phosphate sorption up to about 5 to 20 mmol kg^?1 at all pH, and it induced the soil dispersion, because phosphate sorption increased the negative charge of the soil. After its peak, it decreased with increasing phosphate sorption because the electrolyte concentration increased and the electrolyte screened the electric field near the soil surface. The repulsive potential energy also increased with increasing pH because of the increase of the negative charge of the soil. Even at low pH, after a certain amount of phosphate sorption, the soil dispersed due to the increase of repulsive potential energy, although the soil flocculated before phosphate application. Because the soil dispersion causes soil and phosphorus loss, the influence of soil pH and phosphate sorption on the soil dispersion should be considered for good soil management.     

Bacteriological studies on the mineralization of soil organic nitrogen in paddy soils

Anaerobes were isolated from both disrupted and undisrupted soil samples incubated under submerged condition. All isolates were divided into 9 groups based on their gram stain and morphological characteristics. Correlation coefficients between the amount of nitrogen mineralized and the number of each group were calculated. A significant correlation was observed only between the amount of nitrogen mineralized and the number of gram positive-rod shaped anaerobic microorganisms (r=0.768 at P=0.05). This result indicates that in this study system, sorne particular anaerobes may play an important role in the mineralization of soil organic nitrogen.     

Bacteriological studies on the mineralization of organic nitrogen in paddy soils

In this report we studied the effects of mechanical grinding of paddy soils on nitrogen mineralization and bacterial number when soils were incubated under a submerged condition after grinding.

Nitrogen mineralization was increased by grinding soil samples as compared with those without grinding. The value of (Nd-Nu)/Nu, where Nd is the amount of ammonia-nitrogen formed by incubation of disrupted samples and Nu those of the undisrupted soils, was in good correlation with clay/humus ratio.

Although significant difference was not observed between the number of aerobic bacteria with undisrupted and disrupted soil samples, higher anaerobic bacterial numbers were found with disrupted than with undisrupted ones.

A significant correlation was also obtained between the amounts of nitrogen mineralization increased by the grinding of soil and the numbers of anaerobic bacteria.     

Fuzzy optimization model for integrated management of total nitrogen loads from distributed point and nonpoint sources in watershed

A fuzzy optimization model is developed to allocate allowable total nitrogen (T-N) loads to distributed nonpoint sources (NPSs) and point sources (PSs) in a watershed for river water quality management using the linear programing technique. The watershed is divided into uniform grid cells on which T-N loads issuing from NPSs such as paddy fields, upland crop fields and cities are controlled. A geographic information system integrated with the digital elevation model facilitates computation of route lengths of surface and subsurface flows from cells to a river running through the watershed. The T-N loads discharged from their sources are assumed to decay, subject to distance-related first-order kinetics. As management goals, maximizations of total allowable NPS loads, total allowable PS loads and total yield of rice are considered from environmental and economic viewpoints. A prime constraint is an effluent limitation standard for the aggregate amount of loads that arrive at the downstream end of the river. The fuzzy sets theory helps appropriately describe vague attitudes of decision-makers (i.e., stakeholders and management authorities) in terms of constraints and conflicting goals. An application of the fuzzy optimization model, developed as an improvement over our last nonfuzzy model, to a real watershed in Shiga prefecture, Japan, demonstrates that the fuzzy model embodies our last model, and is capable of creating management alternatives for T-N load allocation in a more practical and flexible manner.     

A distributed hydro-environmental watershed model with three-zoned cell profiling

A cell-based distributed watershed model is developed which enables us to simulate the hydrological and hydraulic aspects of the watershed in a refined fashion. With three-zoned cell profiling, the model is composed of three sub-models; tank model for a surface water zone, soil moisture model for a surface soil zone, and unconfined shallow groundwater flow model for a subsurface zone. Inclusion of the soil moisture sub-model modified to reroute the infiltration, routed from the tank sub-model, into the return flow and the groundwater recharge features the model. The groundwater flow sub-model, numerically approximated by use of the finite volume method and the implicit time-marching scheme, considers a network of on-farm drainage canals as internal boundaries, which is an essential need for modeling the watershed including farmlands. Cascade-linking of the three sub-models in a cell and assembling of all the cells over the entire watershed domain provides the global equations system to be solved. Applicability of the model is demonstrated with its practical application to a real watershed in that paddy and upland crop fields take great part of the land-use practice. It is then indicated in a quantified manner that rice farming significantly contribute as a major groundwater recharger in an irrigation period to fostering and conservation of regional water resources. Along with appropriately profiling a cell, the model is so versatile and tough that it can be applied without difficulty to a watershed of diverse terrains and land-uses and the computations can stably be carried out. It is thus concluded that the model presently developed could be a powerful “watershed simulator” to investigate and assess the time-varying hydro-environmental properties of a watershed while separating and integrating the hydrological and hydraulic components of particular importance.     

Effects of topography and planted trees on the distribution of naturally regenerated broad-leaved trees in a 140-year-old <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> plantation in northern Kyushu,Japan

We investigated factors affecting the distribution of naturally regenerated broad-leaved trees in a 140-year-old Cryptomeria japonica plantation. We used path analysis to examine the relationship among microtopography, the biomass of planted trees, and the biomasses of canopy and subcanopy trees of broad-leaved species. The study plot was divided into three topographic types (ridge, slope, and valley), and we discuss how the different topographic types are affected. For all topographic types, the biomass of canopy trees of broad-leaved species decreased with convexity. For slope and valley topographies, the biomass of subcanopy trees of broad-leaved species also decreased with convexity. For ridge topography, the biomass of subcanopy trees of broad-leaved species increased with the biomass of planted trees, and decreased with the biomass of canopy trees of broad-leaved species. These results suggest the effects of microtopography on the biomass of subcanopy trees were much larger than the effects of canopy trees for slope and valley topographies, while the effects of microtopography were smaller for ridge topography.     

Anatomical characterization of decayed wood in standing light red meranti and identification of the fungi isolated from the decayed area

To further our understanding of wood decay in living light red meranti (Shorea smithiana) trees, microscopic characteristics of the cell and cell wall degradations of S. smithiana wood in the presence of the decay fungi, the identity of the causal fungi, and the decay potential and pattern by an isolated fungus were investigated. Cell wall degradations, including cell wall thinning, bore holes formation, rounded pit erosion, and eroded channel opening were clearly observed under light and scanning electron microscopy. In transverse view, many large voids resulting from a coalition of degraded wood tissue appeared in the decayed canker zone. All these observations suggest the well-known simultaneous decay pattern caused by white-rot fungi. By phylogenetic analysis based on the sequences of internal transcribed spacer region of ribosomal DNA, a basidiomycete fungus isolated from the decayed wood was identified as Schizophyllum commune. The degradation caused by this fungus on sound S. smithiana wood in an in situ laboratory decay test was classified as the early stage of simultaneous decay, and showed a similar pattern to that observed in the wood samples naturally decayed.     

Contribution of a liana species, <Emphasis Type="Italic">Mucuna macrocarpa</Emphasis> Wall., to litterfall production and nitrogen input in a subtropical evergreen broad-leaved forest

Annual amounts of litterfall and nitrogen input by litterfall were measured in a subtropical evergreen broad-leaved forest to examine the contribution of a liana species, Mucuna macrocarpa Wall., to the spatial heterogeneity of litterfall production and nitrogen input. The total litterfall in the study plot was 7.1 t ha⁻¹ year⁻¹. The amount of litterfall varied with topography and was greatest at the valley bottom and decreased toward the ridges. Macuna macrocarpa litterfall was absent on the ridges although it accounted for the largest percentage, 32%, of total leaf litter production in the valley. Nitrogen input by litterfall was 69 kg ha⁻¹ year⁻¹ in the plot. Nitrogen input by litterfall was also largest at the valley bottom and decreased toward the ridges. Leaf litter of M. macrocarpa had approximately twice the nitrogen concentration of litterfall of other species. Macuna macrocarpa accounted for 42% of nitrogen input by leaf litter in the valley. The abundance and the high nitrogen concentration of M. macrocarpa intensified differences in the amount of litterfall and nitrogen input by litterfall between valleys and ridges. It was concluded that a liana species, M. macrocarpa, can contribute to the spatial heterogeneity of litterfall and may subsequently affect nutrient cycling in a subtropical evergreen broad-leaved forest on Okinawa Island.