Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai) by anther culture

Triploid plants were grown from the diploid pear scion cultivar (Pyrus pyrifolia Nakai) by anther culture. Androgenic callogenesis occurred at about 10 days after anther excision, and most calli formed within four weeks and stopped growing at less than 1–3 mm in diameter. Androgenic embryogenesis occurred within 8–12 weeks after anther culture. The formed embryos were generally white, globular or heart-shaped. To increase callogenesis and embryogenesis, the effects of cold pre-treatment, light and activated charcoal (AC) on anthers collected from buds were investigated. Cold pre-treatment, dark and lack of AC increased callogenesis more than no cold pre-treatment, light and adding AC. A decisive condition was not found for embryogenesis. The embryogenesis frequency was extremely low and only six embryos were grown. Two embryos expanded greenish cotyledons and developed foliage, and they proliferated when transferred monthly to fresh proliferation medium. The proliferated shoots were rooted and acclimated. Flow cytometric analysis and investigation of the number of chromosomes of regenerants showed that all cells were triploid.     

Estimation of genetic data and breeding values of traits related to wax production in <Emphasis Type="Italic">Rhus succedanea</Emphasis> L. clones using the REML/BLUP method

Rhus succedanea L. is cultivated in Japan for the wax that can be extracted from its fruits. We determined genetic data for specific traits associated with wax production (fruit number per cluster, FN, fruit weight, FW, number of clusters per area, CN, and wax content, WC) in five traditional cultivars and 13 newer clones. Data were collected over 4 years and alternate bearing was observed: 2001 and 2003 were years with high yields; in 2002 and 2004 yields were poor. The restricted maximum likelihood method was used to calculate (co)variance components for analysis. Broad sense heritability was estimated and ranged from low (FN and CN) to high (FW and WC) for the traits examined. There were positive genetic correlations between FW and WC and negative ones between FN and FW throughout the 4 years. Genetic correlations between CN and the other traits were positive in good crop years and negative in bad crop years. For each assessed clone, the four traits in the 4 years were evaluated using best linear unbiased predictors (BLUP) of the clonal breeding values. The BLUP scores for both FW and WC exhibited positive correlations between pairs of years, whereas there were positive BLUP correlations for CN when the high yield and low yield years were examined separately. The traditional cultivars and the newer clones were also characterized. Some of the clones were better than the traditional cultivars with regard to wax yield and reliability of production. The trait characteristics and the future breeding are discussed.     

Databases on food phytochemicals and their health-promoting effects

Considerable information on the chemistry and biological properties of dietary phytochemicals has accumulated over the past three decades. The scattering of the data in tens of thousands publications and the diversity of experimental approaches and reporting formats all make the exploitation of this information very difficult. Some of the data have been collected and stored in electronic databases so that they can be automatically updated and retrieved. These databases will be particularly important in the evaluation of the effects on health of phytochemicals and in facilitating the exploitation of nutrigenomic data. The content of over 50 databases on chemical structures, spectra, metabolic pathways in plants, occurrence and concentrations in foods, metabolism in humans and animals, biological properties, and effects on health or surrogate markers of health is reviewed. Limits of these databases are emphasized, and needs and recommendations for future developments are underscored. More investments in the construction of databases on phytochemicals and their effects on health are clearly needed. They should greatly contribute to the success of future research in this field.     

Spatial variability of soil solution chemistry under Hinoki cypress (Chamaecyparis obtusa) in Tama Hills

A monitoring study on precipitation and soil solution was conducted to analyze soil acidification processes at the Rolling Land Laboratory (RLL), Hachioji, Tokyo based on the spatial variability of the soil solution chemistry around the Hinoki cypress (Chamaecyparis obtusa) trunk. Soil solution samples were taken at various distances from the tree trunks and at various depths. Soil solution pH at the depth of 10 cm decreased to 4.1–4.2 on the downslope side of large tree trunks, presumably due to the heterogeneity of throughfall input and extensive infiltration of acidic stemflow. Ammonium ions brought by throughfall and stemflow were nitrified and provided large amounts of H⁺. Protons were replaced with exchangeable cations. When base cations were depleted, aluminum ion became the dominant cation species. On the average, Ca²⁺ concentration in the soil solutions at the depth of 10 cm decreased from 0.28 mmol_c L^-1 at the reference site to 0.18 mmol_c L^-1 on the downslope side and Mg²⁺ concentration decreased from 0.30 mmol_c L^-1 to 0.15 mmol_c L^-1. Arithmetic mean aluminum concentration at the depth of 10 cm on the downslope side was 0.35 mmol_c L^-1. Here aluminum dissolution was the main acid sink. Based on the spatial variability of the soil solution chemistry, soil solution acidification processes were divided into four stages.     

Granulomatous nephritis and meningoencephalomyelitis caused by Halicephalobus gingivalis in a pony gelding

A ten-year-old Shetland pony gelding showed low appetite, ataxia, peculiar swaying, clouding of consciousness, and ultimately died. At necropsy, multiple coalescing granulomatous foci were detected in the kidneys, and small necrotic lesions were found in the cerebellum. Histologic examination of the renal tissue sections revealed extensive granuloma, and Halicephalobus gingivalis-like nematodes were seen. Similar nematodes were found in the granulomatous or necrotic lesions of the renal lymph nodes and cerebellum, and were also frequently detected in cerebrospinal meningovascular lesions. Morphologic features together with partial ribosomal RNA gene sequences of the nematodes in the lesions revealed that they were H. gingivalis. The present results indicated that H. gingivalis caused granulomatous nephritis and meningoencephalomyelitis in this pony gelding.     

Serologic and pathogenetic studies on avian reoviruses isolated in Japan

Eighty-nine avian reoviruses isolated from diseased and clinically normal chickens were classified serologically using antisera against five prototype strains. Eighty-three strains were classified into five serotypes; six strains were untypable. Most of the cytopathogenic strains that produced a clear cytopathic effect (CPE) in chicken embryo fibroblasts (CEFs) were highly pathogenic for chicken embryos (80% or more mortality via the allantoic sac) and for chicks (severe footpad swellings and tenosynovitis). These strains were classified into a single serotype represented by the TS-142 prototype strain. However, 10 strains that could not produce a clear CPE in CEFs showed very low pathogenicity for embryos and chicks, and these strains were serologically different from the TS-142 prototype strain. There was a strong relationship between pathogenicity and serotype. About 17% of the isolates were considered highly pathogenic.     

The use of tetracycline in anthelmintic baits to assess baiting rate and drug efficacy against Echinococcus multilocularis in foxes

Anthelmintic (praziquantel) baiting of wild red foxes against Echinococcus multilocularis infection was studied in a highly epizootic suburban area of Otaru, Hokkaido (the northern island of Japan) during the summer and autumn in the years 1999-2004. Acceptance of baits containing the biomarker tetracycline (TC) was evaluated. The prevalence of E. multilocularis infection in foxes before baiting (1999-2000) was 58% (88/153), whereas in the fourth year of bait distribution year (2004), it decreased to 11% (5/45). Analysis of TC marking in the teeth of foxes showed that 39% (77/195) of those captured after baiting were estimated to have consumed baits in the year of capture. Importantly, more juvenile (56%, 49/87) than adult foxes (26%, 28/108) were marked, indicating efficient baiting of juveniles, which tended to have a higher worm burden of E. multilocularis. Of 77 marked foxes, E. multilocularis and Alaria alata (monitored as the second indicator species of deworming) were not detected in 70 (90%) and 76 (99%) foxes, respectively. The results suggest effective deworming by bait consumption. However, it was also demonstrated that 9% of the marked foxes were infected or re-infected after bait consumption, suggesting high infection pressure and the importance of frequent baiting.     

Sugar-end in Russet Burbank potatoes

Tissue selected from the stem- and bud-end of reconditioned tubers having sugar-end disorder was compared by color after frying and chemical composition to tissue removed from the two ends of tubers without sugar-end disorder taken directly from cold storage at 45 F, and after reconditioning 3 weeks at 70 F. Chemical analyses of selected tissue suggested that the dark finish-fry color at the stem-end of sugar-end tubers following reconditioning at 70 F was due to the inability of the physiological systems within the tissue to convert reducing sugars to starch and/or to use these sugars in the respiratory process. Of the components analyzed, only a high reducing sugar content was associated with a dark finish-fry color. Neither sucrose nor free amino acid content was associated with the sugar-end disorder. Extending the reconditioning period at 70 F for sugar-end tubers did not lower the reducing sugar content or the darkening of the stem-end tissue on frying.     

Effectiveness of tetrazolium salts in microbial ecological studies in submerged soil

Several trials to obtain information on the living microorganisms at micro-sites of the submerged soil by the novel utilization of the tetrazolium salts were described. Results are summarized as follows:

(1) The air-dried soil was incubated with TIC (triphenyl tetrazolium chloride) under submerged condition according to the modified double layer plate method. The red formazan (the reduced product of TIC) was precipitated at the biologically active sites by the dehy. drogenase activity. Using a binocular microscope, most of the biologically active sites were found to be localized on and around the organic debris.

(2) The amount of nitrogen mineralized by incubating the soil under submerged condi. tion correlated well not only with the amount of formazan formed biologically in the submerged soil but also with the amount of formazan formed non-biologically by heating the soil with TIC in Ba(OH). solution. In addition, the pattern of precipitation of the nonbiologically formed formazan was very similar to that of the biologically formed formazan. Accordingly, the substances which reduced TIC non-biologically were considered to be closely associated with the substrates for the microorganisms.

(3) The plant debris stained with the red formazan could be mounted on a slide glass and observed at high magnification for the detailed study on the process of decomposition. Addition of lactophenol cotton blue to the plant debris could visualize the microorganisms responsible for the decomposition of the plant debris.

(4) NTB (nitro tetrazolium blue) was the most suitable for staining individual microorganisms growing on cellulosic materials or plant debris. The double staining with NTB and rosebengal made the discrimination between dead or resting cells and viable ones possible when this technique was applied to the decomposed filter paper.     

Organic matter production in rice field flood water

To assess the contribution of organic matter produced in the flood water to the fertility of a rice soil, the primary productivity and the algal biomass therein were examined throughout one crop. Primary productivity was estimated from the diurnal curve of dissolved oxygen.

Just after transplanting, an algal bloom developed due to fert,ilizer or ploughing or both. After submerged weeds occupied the whole paddy no distinct algal growth was found. At the ripening stage, the rice plant canopy suppressed the growth of aquatic plants. Benthic algal biomass did not change much throughout the crop period. The standing crop of algae ranged from 2 to 114 kg/ha by fresh weight, while the maximum standing crop of submerged weeds (Najas sp., Chara sp.) was 400 kg/ha by dry weight.

The primary productivity of the flood water community was high (0.6-3.3 g 0 1 m^-2day^-l) and equivalent to productivity values in eutrophic lakes. The total gross primary production of the flood water community during the cropping period corresponded to 10% and 15% of that of rice plant in the fertilized plot and non-fertilized plot, respectively.

Considering the movement of CO₂ in the flood water, it is suggested that the photosynthesis activity in the flood water prevents surplus CO₂ from being lost.