Polyacrylamide gel electrophoresis of S-RNase fragments for identification of S-genotypes of Japanese pear (Pyrus pyrifolia)

Japanese pear (Pyrus pyrifolia) exhibits gametophytic self-incompatibility (GSI) controlled by a complex and multiallelic S locus. The pistil-part product of the S locus is the polymorphic ribonuclease, S-RNase. Information on S-genotypes is important for the production and breeding of Japanese pears. Molecular analyses of S-genotypes of Japanese pear have been conducted with the CAPS (cleaved amplified polymorphic sequence) system; PCR amplification of S-RNase fragments by a common primer pair followed by digestion with restriction enzymes each of which cleaves a specific S haplotype. Here, we show that the separation of S-RNase fragments by polyacrylamide gel electrophoresis (PAGE) distinguishes four out of nine S haplotypes of Japanese pear without restriction digestion. S³-, S⁵-, S⁶- and S⁸-RNases were identified as distinct bands by PAGE. S³- and S⁵-RNases were separated by PAGE despite their identical fragment sizes. Using this system, three Japanese pear lines with unknown S-genotypes were analyzed. The newly determined S-genotypes of the lines were confirmed by CAPS analysis.     

Alteration in the apoptosis process of rat esophageal epithelium with hyperproliferation of indigenous bacteria under a physiological condition

The apoptosis process in rat esophageal epithelium was investigated using enzyme-immunohistochemistry and transmission electron microscopy. As a result, Fas and Fas-L were expressed in the epithelial cell membrane and cytoplasm from the stratum spinosum (SS) to the stratum granulosum (SG). No TNF-R1 show immunopositivity in the cell membranes. TNF-α and caspase-8 were not observed in any layer. Caspase-10, cleaved caspase-3, XIAP and DNase-1 were found in the epithelial cytoplasm from the SS to the SG, whereas Bid, Apaf-1 and cleaved caspase-9 were detected only in the SG. Cytochrome c was observed as cytoplasmic granular positivity from the stratum basale (SB) and altered into homogeneous immunopositivity in the SG. Bcl-2 and Bcl-X immunopositivity was detected in cytoplasm from the SB to the SG. Immunoreactions of Bak in the cytoplasm and Bax beneath the cell membrane were observed from the upper portion of the SS with increasing intensity toward the SG. In the sites with the hyperproliferation of indigenous bacteria, TNF-R1, TNF-α and caspase-8 were detected in the SG and the immunopositive intensities of Bid, Apaf-1 and cleaved caspase-9 were altered to be strong. Prominently swollen cells and decreased mitochondria were ultrastructurally confirmed in the uppermost layers of stratum corneum. These findings suggest that the Fas-Fas-L-interaction initially induces apoptosis through a mitochondria-independent pathway and secondarily through a mitochondria-dependent pathway, leading to eventual epithelial cell death in the rat esophageal epithelium. The bacterial stimuli probably enhance the mitochondria-dependent pathway through the TNF-R1-TNF-α interaction.     

Gas-phase pyrolysis of methyl glucosides and levoglucosan

Levoglucosan, the major intermediate in cellulose pyrolysis, is stable up to around 500?°C in the gas phase. To study whether this stability is a characteristic property of levoglucosan, the gas-phase reactivity was compared with those of methyl α- and β-glucosides at 200–500?°C (residence time 1.2–2.0 s). The methyl glucosides decomposed even at 200 and 300?°C to form levoglucosan exclusively. This selective transglycosylation was explained with a concerted mechanism. Fragmentation of the glucosides forming C1–C3 carbonyl compounds started at 400?°C, a temperature lower than that of levoglucosan (500?°C). Thus, levoglucosan is a special carbohydrate that is stable in the gas phase, and the stability is explained by the steric hindrance of the bicyclic ring. Formation of the anhydrofuranose isomer and furans was negligible from the gas-phase pyrolysis of these compounds, suggesting that these are produced mainly from the molten-phase pyrolysis. These results show the roles of gas- and molten-phase reactions during carbohydrate pyrolysis, providing insights for upgrading biomass pyrolysis/gasification processes.     

Lignin pyrolysis reactions

Lignin, an aromatic constituent of woody biomass, is a potential renewable aromatic feedstock for a sustainable future carbon economy. Pyrolysis-based technologies, such as fast pyrolysis and gasification, are promising methods for converting lignin into biochemicals, biomaterials, and biofuels. A better understanding of the molecular mechanisms involved in lignin pyrolysis/gasification would guide the development of the controlled pyrolysis and gasification systems to overcome issues with low product selectivity, an intrinsic drawback of current pyrolysis-based technologies. This review article summaries the state-of-the-art research into molecular mechanisms of lignin pyrolysis and gasification. This information should also be useful for understanding the influence of high temperature heat treatments on the properties of wood.     

Bartonella species in wild rodents and fleas from them in Japan

The purpose of this study was to assess the role of fleas for transmission of Bartonella species among wild rodents in Japan. Flea samples were collected from wild rodents and examined genetically for Bartonella infection. Bartonella DNA was detected from 16 of 40 (40.0%) flea samples. Sequence analysis demonstrated that 3 of 16 (18.8%) of the Bartonella-positive animals were infested with fleas from which the closely related Bartonella DNA sequence was detected, indicating that the fleas acquired Bartonella from the infested rodents. The DNA was detected in hemolymph, the midgut and the ovary (only in female), indicating that Bartonella might be colonized through the midgut and distributed into the body.     

Application of N2/Ar ratio method for N2 fixation studies in submerged soil

Abstract

Recently there has been developments in the measurement of N₂ fixation due mainly to the C₂H₂ reduction method (1). This method, however, has several disadvantages, especially for submerged soil, and the estimated amount of fixed N₂ on the basis of the C₂H₂ reduction activity is not very reliable. The tracer ¹⁵N₂ technique which gives a reliable estimation of the fixed N₂ is too expensive for common use. Development of an alternative method suitable for submerged soil would therefore be desirable. The present authors expected that the measurement of the ratio N₂/Ar in the soil solution might provide advantages for the estimation of the fixed N₂ in submerged soil.     

The relationship of readily decomposable organic matter of the paddy soil to its colloidal complexes

The soil nitrogen of paddy soil is found to exist in the form of organic and inorganic nitrogen. In general, the content of inorganic nitrogen is so low that it is mainly occupied by organic nitrogen. Such soil organic nitrogen is characterized by large resistance to decomposition by soil microorganisms, and so its mineralization occurs to a small extent in most cases. However, as revealed by M. Shioiri¹⁾ and T. Harada et al. ^2,3), by some appropriate treatment of paddy soil, a part of soil organic nitrogen becomes decomposed by microorganisms and mineralized under flooded condition. Among various treatment, for example, are Presented the followings: (1) air-drying of soil Prior to flooding; (2) raising the soil temperature during the flooded condition; (3) raising the soil reaction (pH 9.0) by addition of weak alkali solution; (4) the addition of neutral salts such as Nafiuoride, Na-phosphate, Na-oxalate etc., which make soil humus peptizable.     

Studies on the organo-mineral colloidal complexes of paddy soil (Part 2) Humus contained in G1 of the degraded paddy soil

Regarding paddy soil colloids as the so-called “organo-mineral colloidal complexes” of A. F. Tyulin, the authors separated the colloidal fractions after his method, and reported the characrzstics of each fraction, and content of humus and some characteristics thereof in the previous paper¹⁾.     

Phosphorus (P) Uptake mechanisms of crops grown in soils with low P status

In our previous studies, pigeonpea (Cajanus cajan L.), groundnut (Arachis hypogaea L.), and rice (Oryza sativa L.) were found to have a higher ability to take up Fe- or Al-bound phosphorus (P) than soybean (Glycine max L.) and sorghum (Sorghum bicolor L.). Phosphorus absorption characteristics like I _max, K _m, C _min, and Fe^III reduction activity of roots, and root exudates in various crops were examined with a view to analyzing the mechanisms of P uptake. Phosphorus uptake ability was largely unrelated to variations in I _max, K _m, C _min, and Fe^III reduction activity of roots. Phosphorus-solubilizing activity in anionic fractions of root exudates was detected in pigeonpea but not in rice or groundnut. Malonic acid was the major component followed by oxalic and piscidic acid. These organic acids were able to release P from FePO₄ and A1PO₄. The higher P uptake ability of pigeonpea in soils with low P fertility presumably depends on the secretion of such organic acids from roots.