Soft rot of root chicory in Hokkaido and its causal bacteria

In August 2010, bacterial soft rot was found on root chicory (Cichorium intybus var. sativum) in Hokkaido, Japan. Severely infected plants in fields were discolored, had wilted foliage, and black necrosis of petioles near the crown. Wilted leaves subsequently collapsed and died, forming a dry, brown or black rosette. The root and crown became partially or wholly soft-rotted. Slimy masses on infected areas of roots, turned dark brown or black. Gram-negative, rod-shaped, peritrichously flagellated, facultatively anaerobic bacteria were exclusively isolated from rotted roots, and typical symptoms were reproduced after inoculation with the strains. The bacteria were identified as Dickeya dianthicola, Pectobacterium carotovorum subsp. carotovorum, and Pectobacterium carotovorum subsp. odoriferum based on further bacteriological characterization and the sequence analysis of the malate dehydrogenase gene and 16S rRNA gene. These bacteria should be included with the previously reported Dickeya (=Erwinia) chrysanthemi in Saitama Prefecture, Japan, as causal pathogens of bacterial wilt of chicory.     

Pathogenicity and immunogenicity of equine herpesvirus type 1 mutants defective in either gI or gE gene in murine and hamster models

To develop a live vaccine for equine herpesvirus type 1 (EHV-1), two EHV-1 mutants containing no heterogeneous DNA, DeltagI and DeltagE, were constructed with deletions in the open reading frame of either glycoprotein I (gI) or E (gE), respectively. In equine cell culture, deletion mutants formed smaller plaques than the parental and revertant viruses, but the one-step growth patterns of the deletion mutants and the parental strain were approximately the same. These results suggest that both gI and gE contribute to the ability of EHV-1 to spread directly from cell-to-cell, but that these glycoproteins are not required for viral growth in vitro. Mice and hamsters inoculated intranasally with these mutants showed no clinical signs, and continued to gain weight, whereas those inoculated with the parental virus exhibited a reduction in mean body weight. Furthermore, nervous manifestations were observed in hamsters inoculated with the parental virus. These results suggest that gI and gE have an important role in EHV-1 virulence including neurovirulence in experimental animal models. On the other hand, serum neutralizing antibodies were detected in mice immunized with DeltagI or DeltagE at two weeks after inoculation. Following challenge with the parental virus, DeltagI- or DeltagE-immunized mice were able to clear parental virus from their lungs faster than mock-immunized mice. These results suggest that the EHV-1 mutants defective in gI and in gE are attenuated but have ability to elicit immune responses in inoculated mice that contribute to virus clearance.     

In vitro efficacies of oseltamivir carboxylate and zanamivir against equine influenza A viruses

To investigate the possibilities of two NA inhibitors [oseltamivir carboxylate (OC) and zanamivir (ZA)] as the clinical agents for equine influenza A virus (EIV) infection, we examined the efficacies of these inhibitors against twelve EIVs in vitro. OC and ZA inhibited NA activities of all EIVs with 50% inhibitory concentrations with ranging from 0.017 to 0.130 and from 0.010 to 0.074 microM, respectively. OC and ZA inhibited plaque-forming of all EIVs in MDCK cells with 50% effective concentrations with ranging from 0.015 to 0.097 and from 0.016 to 0.089 microM, respectively, except for one strain (13.328 microM and 6.729 microM). These results suggest that these inhibitors are effective against most EIVs and might be useful for treatment of EI in horses.     

Biological control of apple crown gall by nonpathogenic Rhizobium vitis strain VAR03-1

A nonpathogenic strain of Rhizobium vitis, VAR03-1, was tested as a biological control agent for crown gall of apple. When apple tree roots were soaked in a cell suspension of antagonists before planting in the field, treatment with VAR03-1 significantly reduced the number of plants with tumors and the disease severity. The inhibitory effects of treatment with VAR03-1 and the nonpathogenic R. rhizogenes strain K84, which is commercially used as a biological control agent throughout the world, on apple crown gall were almost identical. VAR03-1 was bacteriocinogenic, producing a halo of inhibition against two species of tumorigenic Rhizobium isolated from galled apple trees. Moreover, VAR03-1 established populations on roots of apple tree rootstock and persisted on roots for 437?days. This is the first report that the nonpathogenic R. vitis VAR03-1 effectively controls crown gall of apple.     

Detection of QTLs for white-back and basal-white grains caused by high temperature during ripening period in japonica rice

There is increasing evidence that global warming affects the development of rice. High temperatures during ripening increase the ratio of undesirable chalky grains followed by deteriorating grain appearance quality. In order to detect quantitative trait loci (QTLs) controlling the occurrence of white-back and basal-white chalky grains of brown rice, QTL analysis was performed using recombinant inbred lines derived from a cross between two strains, ‘Tsukushiroman’ (sensitive to heat stress) and ‘Chikushi 52’ (tolerant of heat stress). The F₇ and F₈ lines were exposed to heat stress during the ripening period in two locations, Fukuoka and Kagoshima, in Japan. QTLs for white-back grains and basal-white grains were detected on chromosomes 1, 3, and 8, and those for basal-white grains were detected on chromosomes 2, 3, and 12. QTLs on chromosome 8 for white-back grains were shared in the plants grown in both locations. Near-isogenic lines (NILs), which harbored a segment from ‘Chikushi 52’ on chromosome 8 with the genetic background of ‘Tsukushiroman’, showed relatively lower ratios of white-back grains than ‘Tsukushiroman’. Therefore, insertion of the ‘Chikushi 52’ genomic region of the QTL on chromosome 8 can improve the quality of rice when it is grown under heat stress conditions.     

Effects of Plant Species on CH4 and N2O Fluxes from a Volcanic Grassland Soil in Nasu, Japan

To investigate the effects of plant species in grassland on methane (CH₄) and nitrous oxide (N₂O) fluxes from soil, fluxes from an orchardgrass ( Dactylis glomerata L.) grassland, white clover ( Trifolium repens L.) grassland and orchardgrass/white clover mixed grassland were measured weekly from April 2001 to March 2002 using a vented closed chamber method. Related environmental parameters (soil inorganic N content, soil pH (H₂O) value, soil moisture content, soil temperature, grass yield, and the number of soil microorganisms) were also regularly monitored. On an annual basis, CH₄ consumption in the soil of the orchardgrass grassland, white clover grassland and orchardgrass/white clover mixed grassland was 1.8, 2.4, and 1.8 kg C ha⁻¹ year⁻¹, respectively. The soil bulk density of the white clover grassland was lower than that of the other grasslands. Fluxes of CH₄ were positively correlated with the soil moisture content. White clover increased the CH₄ consumption by improving soil aeration. Nitrogen supply to the soil by white clover did not decrease the CH₄ consumption in the soil of our grasslands. On the other hand, annual N₂O emissions from the orchardgrass grassland, white clover grassland, and orchardgrass/white clover mixed grassland were 0.39, 1.59, and 0.67 kg N ha⁻¹ year⁻¹, respectively. Fluxes of N₂O were correlated with the NO₃⁻ content in soil and soil temperature. White clover increased the N₂O emissions by increasing the inorganic N content derived from degrading white clover in soil in summer.     

Study of the role of the carbohydrate and protein moieties of soy soluble polysaccharides in their emulsifying properties

The objective of this work was to investigate the role played by the protein fraction of soy soluble polysaccharide (SSPS) during its adsorption at oil/water interfaces. SSPS was separated in a high (HMF; 310 kDa) and low (LMF; 20 kDa) molecular weight fraction by gel filtration. SSPS/HMF consisted of 91.6% carbohydrate and 2.2% protein and showed better emulsifying properties than those of the whole SSPS, whereas SSPS/LMF seemed to affect negatively the adsorption behavior of SSPS. SDS-PAGE of the protein fraction obtained from SSPS/HMF showed a molecular mass of 50 kDa, was composed predominantly of proline (23.1%) and glutamic acid (15.2%), and still contained 8.8% of neutral sugar and 5.3% of uronic acid. Results indicated that not all of the protein material present in SSPS contributes to SSPS functionality but that only the material associated with HMF aids in the adsorption of SSPS onto oil/water interfaces.