Quantitative nested real-time PCR detection of <Emphasis Type="Italic">Verticillium longisporum</Emphasis> and <Emphasis Type="Italic">V. dahliae</Emphasis> in the soil of cabbage fields

Verticillium longisporum and V. dahliae, causal agents of Verticillium wilt, are spreading through the cabbage fields of Gunma Prefecture. Using the V. longisporum-specific intron within the 18S rDNA and differences between ITS 5.8S rDNA sequences in Japanese isolates of V. longisporum and V. dahliae, we developed three quantitative nested real-time (QNRT) PCR assays. The QNRT-PCR quantification of V. longisporum or V. dahliae in cabbage field soil was consistent with the severity of Verticillium wilt disease in those fields. In field trials of resistant cultivar YR Ranpo grown for three seasons in soil infested with the pathogen, disease severity and pathogen density in the soil were significantly reduced in a field moderately contaminated by V. dahliae, but only slightly reduced in a highly contaminated field. These results suggest that continuous cultivation of a resistant cultivar is an effective way to reduce the pathogen population. QNRT-PCR assays provide a powerful analytical tool to evaluate the soil population dynamics of V. longisporum and V. dahliae for disease management.     

Comparison of Bacterial Communities Associated with Microcrustaceans from the Floodwater of a Paddy Field Microcosm: Estimation Based on DGGE Pattern and Sequence Analyses

Bacterial communities associated with five kinds of microcrustaceans ( Tanycypris sp., Moina sp., Mesocyclops sp., Cypretta sp. and Heterocypris sp.) from the floodwater of a paddy field microcosm were examined by the application of denaturing gradient gel electrophoresis (DGGE) to PCR-amplified 16S rDNA products with universal bacterial primers and by sequencing of characteristic DGGE bands. The number of DGGE bands of the associated bacteria was small, indicating the association of specific bacterial members with the microcrustaceans studied, among which Tanycypris sp. showed the smallest number of bands. Principal component analysis (PCA) demonstrated that the community structure of the associated bacteria could be divided into three groups: Podocopida ( Tanycypris sp., Cypretta sp. and Heterocypris sp.), Moina sp. and Mesocyclops sp., and further analysis separated Tanycypris sp. and Heterocypris sp. into different clusters. The duration of the incubation period affected the bacteria associated with Tanycypris sp., Moina sp. and Cypretta sp. only. Nearly all of the associated bacteria belonged to Gram-negative bacteria, especially the Cytophaga-Flavobacterium-Bacteroides (CFB) group. Closest relatives of the DGGE bands common to three Podocopida and Mesocyclops sp. belonged to an invertebrate endosymbiont.     

Carbon cycling in rice field ecosystems in the context of input, decomposition and translocation of organic materials and the fates of their end products (CO2 and CH4)

Rice fields are intensively managed, unique agroecosystems, where soil flooding is general performance for rice cultivation. Flooding the field results in reductive soil conditions, under which decomposition of organic materials proceeds during the period of rice cultivation. A large variety of organic materials are incorporated into rice soils according to field management. In this review, the kind and abundance of organic materials entering carbon cycling in the rice field ecosystem are evaluated first. Then, decomposition of plant residues and soil organic matter in rice fields is reviewed quantitatively. Decomposition of plant residues is shown to be the active process in carbon cycling in rice fields. Rice releases photosynthates into the rhizosphere (rhizodeposition), and they follow a different avenue of decomposition in soil from that of plant residues. Incorporation of rhizodeposition into microbial biomass and soil organic matter during the period of rice cultivation, and their fates after harvesting are evaluated quantitatively from ¹³C pulse labeled experiments. Percolating water transports inorganic and organic carbon from the plow layer to the subsoil layer. The amounts of their transport and accumulation in the subsoil layer are evaluated in relation to the amounts of soil organic C in the plow layer. Not only CO₂ but also CH₄ are produced in the decomposition process of organic materials in flooded rice fields. CH₄ evolution from rice fields is of global concern from the viewpoint of global warming. Origins of CH₄ evolved from rice fields are estimated first, followed by the fates of CH₄ in rice field ecosystems. Rhizodeposition is shown to be the main origin of CH₄ evolved from rice fields. Evolution to the atmosphere is not the sole pathway of CH₄ produced in rice fields. The amounts of CH₄ retained in soil, percolated to the subsoil layer and decomposed in soil are evaluated in the context of the amounts of CH₄ efflux. Thus, this review focuses on carbon cycling in the rice field ecosystem from the viewpoints of input, decomposition, and translocation of organic materials and the fates of their end products (CO₂ and CH₄).     

Pressure-induced denaturation of monomer beta-lactoglobulin is partially irreversible: comparison of monomer form (highly acidic pH) with dimer form (neutral pH)

This study was conducted to assess the effect of high hydrostatic pressure on monomer beta-lactoglobulin (BLg) at acid pH by fluorescence spectroscopy under pressure and by circular dichroism (CD) and (1)H NMR spectroscopies after release of pressure. The intrinsic (tryptophan) fluorescence measurement and the study of 8-anilinonaphthalene-1-sulfonate (ANS) binding to BLg indicated that at pH 2.0 the recovery of center of spectral mass or ANS fluorescence was almost complete upon pressure release. No difference in (1)H NMR spectra was observed between pressurized and unpressurized BLg. In addition, NMR detection of the H/D exchange of aromatic protein indicated that the conformation of the vicinity of tryptophan residues could be refolded almost completely after release of pressure. These results seemingly confirm that the pressure-induced denaturation of BLg at pH 2.0 is reversible. However, cis-parinaric acid binding ability of pressurized BLg was largely lost, although its retinol binding ability was the same as its unpressurized one. Furthermore, CD spectra of the far-UV region and 2D NMR spectra evidently revealed the difference in the conformation of the molecule between unpressurized and pressurized BLg. These results are interpreted as an existence of partially fragile structure in the BLg molecule by high pressure.     

Study on some functional and compositional properties of yak butter lipid

The present experiment was performed to determine some functional properties of yak butter lipids such as lipid class composition; conjugated linoleic acid (CLA) composition, differential scanning calorimetric (DSC) analysis, tyrosinase inhibition activity and antioxidant property. Yak butter lipids composition contained 98% triacylglycerols, 0.9% free fatty acids, 0.32% free sterols and 0.27% phospholipids. The CLA content in yak butter was 2.5% and the major portion was of cis‐9 and trans‐11 (90%). The DSC analysis of yak butter lipids showed a similar path for transition temperature as cow butter lipids, although the enthalpy of yak lipid was higher (40.0 mJ/mg) compared to cow butter lipids (32.0 mJ/mg). Melting point of yak butter was observed at 41°C. Yak butter with lactic acid, NaCl, citric acid and ascorbic acid showed pronounced tyrosinase inhibition activity. Vegetable oils blended with yak butter have extended the oxidation induction time.     

Humoral immune response to non-viable Mycoplasma pulmonis in mice enhanced by dextran sulfate

The enhancing effect of dextran sulfate on the humoral immune response to nonviable Mycoplasma pulmonis in mice was evaluated by means of the indirect haemagglutination test. The serum antibody titres in mice immunized subcutaneously with a mixture of non-viable M. pulmonis and dextran sulfate were greater and persisted longer than those in mice immunized with non-viable M. pulmonis alone. DEAE-dextran also enhanced the humoral immune response to non-viable M. pulmonis in mice.