Construction of a system for exploring mitotic homologous recombination in the genome of Pyricularia oryzae

To investigate mitotic homologous recombination (HR) in Pyricularia oryzae, we created an HR detection system. The system consists of two non-functional enhanced yellow fluorescent protein (YFP) and blasticidin S deaminase (BSD) fusion genes (YFP::BSD). If mitotic HR occurs between the two non-functional genes in the genome, restoration of the functional YFP::BSD gene can be expected. The expression of the functional YFP::BSD gene can be detected by both YFP fluorescence and resistance against BS. When the P. oryzae genome was transformed simultaneously with two non-functional genes, all six lines of transformants with both genes had some portion of their hyphae exhibiting YFP fluorescence and BS resistance during growth. Up to ca. 10 % of conidia harvested from the mycelium of each of the six lines had YFP fluorescence, suggesting that HR consistently occurs during mycelium growth. To determine whether and how the HR-mediated phenotypic changes occurred at the DNA level, we analyzed the genomic DNA of BS-resistant mycelia by PCR-RFLP and sequencing and were able to confirm the existence of a restored functional YFP::BSD gene and a non-functional recombinant gene. Collectively, these findings demonstrate that we established a successful HR detection system for P. oryzae, which can be used for other plant pathogens, and that mitotic HR actually occurs in P. oryzae and constitute the first experimental evidence for mitotic HR in a fungus.     

Classification of the genus Bacillus based on MALDI-TOF MS analysis of ribosomal proteins coded in S10 and spc operons

A rapid bacterial identification method by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) using ribosomal proteins coded in S10 and spc operons as biomarkers, named the S10-GERMS (the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum) method, was applied for the genus Bacillus a Gram-positive bacterium. The S10-GERMS method could successfully distinguish the difference between B. subtilis subsp. subtilis NBRC 13719(T) and B. subtilis subsp. spizizenii NBRC 101239(T) because of the mass difference of 2 ribosomal subunit proteins, despite the difference of only 2 bases in the 16S rRNA gene between them. The 8 selected reliable and reproducible ribosomal subunit proteins without disturbance of S/N level on MALDI-TOF MS analysis, S10, S14, S19, L18, L22, L24, L29, and L30, coded in S10 and spc operons were significantly useful biomarkers for rapid bacterial classification at species and strain levels by the S10-GERMS method of genus Bacillus strains without purification of ribosomal proteins.     

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 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.     

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.     

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.     

Physiological characteristics of blue sac disease in chum salmon alevins

Fisheries Science - Blue sac disease (BSD) in salmonids results in the abnormal accumulation of ascites fluid and can often lead to mass mortality. However, the symptoms and causes of BSD remain...     

Inhibition by pulp juice and enhancement by ingestion on germination of bird-dispersedPrunus seeds

To evaluate what prevents seed germination and why ingestion by birds enhances germination, we carried out germination experiments using bird-dispersed seeds of twoPrunus species (Prunus sargentii andP. ssiori) in the laboratory. Germination of seeds with following four treatments was compared: “Ingested seeds” excreted after feeding of fruits to birds; “Extracted seeds” deliberately extracted from the fruit pulp; “Juiced seeds” which were supplemented with the juice of the pulp after seeds had been deliberately extracted from the pulp; and “Intact seeds” in untreated whole fruits. Many ingested (27%, 23% inP. sargentii, 18% inP. ssiori) and extracted seeds (24% inP. sargentii, 17% inP. ssiori) germinated, and difference in germination percentages between ingested and extracted seeds were not significant, for eitherPrunus species. The Juiced seeds rarely germinated (2% inP. sargentii, 6% inP. ssiori), and seeds within intact fruits did not germinate. These results suggest that the fruit pulp and the juice of the pulp inhibit germination and the seeds could germinate without ingestion by birds once they had been manually extracted from pulp. Therefore, it is considered that bird ingestion enahnces germination by removing pulp, especially juice of the pulp, in these twoPrunus species.     

Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2

Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS.     

Nickel-catalyzed carbonization of wood for coproduction of functional carbon and fluid fuels I: production of crystallized mesoporous carbon

Japanese larch wood loaded with nickel (1%–4%) alone or with nickel and calcium (0.25%–1.5%) was carbonized at 800°–900°C for 0–120min with a heating rate of 5°–20°C min⁻¹ in a helium flow of 5.8−46.4 ml STP cm⁻² min⁻¹ to examine the influence of these variables on the crystallization of carbon (the formation of T component) and the development of mesoporosity. From the obtained results, reaction conditions suitable for effective production of carbon with the dual functions of adequate electroconductivity and adsorption capacity in liquid phase were established, thereby explaining the factors that govern the process. It was also confirmed that mesopore having a diameter of about 4 nm was selectively produced at the cost of specific (BET) surface area in parallel with the formation of T component. This result provided good insight into how the simultaneous dual function could be realized.