Use of fluorescent proteins to visualize interactions between the Bakanae disease pathogen <Emphasis Type="Italic">Gibberella fujikuroi</Emphasis> and the biocontrol agent <Emphasis Type="Italic">Talaromyces</Emphasis> sp. KNB-422

Talaromyces sp. isolate KNB-422, isolated from a rice seedling, is a biofungicidal agent effective against several seedborne pathogens of rice including Gibberella fujikuroi, which causes Bakanae disease. Because the fungal mode of action (MOA) has not yet been clarified, we used the fluorescent protein markers GFP and RFP to visualize cell–cell interactions between the biocontrol agent and the pathogen G. fujikuroi. In slide culture, the hyphal cell wall of G. fujikuroi collapsed, and fluorescence of its cytoplasm disappeared 3 days after contact with hyphae of Talaromyces sp. On inoculated rice plants, both fungi occupied the same regions of coleoptiles and roots, where the biocontrol effect of Talaromyces sp. must be exerted. Our observations suggest that the MOA of Talaromyces sp. is mycoparasitic.     

Enhancement of callose production by a combination of aluminum and iron in suspension-cultured tobacco (Nicotiana tabacum) cells

In nutrient medium, aluminum (A1) enhances ferrous ion [Fe(II)] -mediated per oxidation of lipids, which results in the loss of the plasma membrane integrity and the accumulation of A1 in tobacco cells. Under these conditions, the mechanism of callose production and possible involvement of callose in the accumulation of Al were investigated. Callose production was enhanced by both Al and Fe(II), but not by A1 or Fe(II) alone, and the enhancement was inhibited by a lipophilic antioxidant, suggesting that the enhancement of callose production is caused by the A1-enhanced, Fe(II)-mediated peroxidation of lipids. The enhancement of callose production depended on the presence of external Ca²⁺ in the treatment medium. The activity of β-l,3-glucan synthase in the microsomes was increased several times by the addition of Ca²⁺ in the assay medium, although the activity in the microsomes was reduced by the treatment of cells with Al and Fe(II) together. Therefore, it is likely that callose production is enhanced by exogenous Ca²⁺ via the AI-enhanced, Fe(II)-mediated peroxidation of lipids. During the exposure of the cells to Al and Fe(II), callose production started and increased simultaneously with Al accumulation. However, the digestion of callose in the cell wall materials prepared from the A1-treated cells by laminarinase did not release A1, suggesting that callose is not involved in the binding or trapping of A1.     

Gross nitrification rates in four Japanese forest soils: heterotrophic versus autotrophic and the regulation factors for the nitrification

Measurements of gross NH ₄ ⁺ and NO ₃ ^? production in forest soils were conducted using the ¹⁵N pool dilution method. Mineral topsoils (0?C10?cm depth) were collected from four forests from northern to southern Japan with a natural climate gradient to elucidate the mechanisms regulating gross nitrification rates in forest soils. Additionally, we attempted to evaluate the relative importance of heterotrophic nitrification in gross total nitrification using acetylene as a specific inhibitor of autotrophic nitrification. Distinct differences were found among sites in the gross rates of NH ₄ ⁺ production (3.1?C11.4?mg?N?kg^?1?day^?1) and gross total nitrification (0.0?C6.1?mg?N?kg^?1?day^?1). The rates of gross heterotrophic nitrification were low in this study, indicating that heterotrophic nitrification is of minor importance in most forest mineral topsoils in Japan. Significant relations were found between gross autotrophic nitrification and gross NH ₄ ⁺ production, soil N, and soil C concentrations, but none was found between gross autotrophic nitrification and soil pH. We determined the critical value of the gross NH ₄ ⁺ production rates for gross autotrophic nitrification under which no gross autotrophic nitrification occurred, as well as the critical soil C/N ratio above which gross autotrophic nitrification ceased. Results show that tight coupling of production and consumption of NH ₄ ⁺ prevents autotrophic nitrifiers from utilizing NH ₄ ⁺ as long as NH ₄ ⁺ availability is low.     

Expression of mRNA for sodium-glucose transporter 1 and fatty acid translocase in the ruminant gastrointestinal tract before and after weaning

The mRNA expression of sodium‐glucose transporter 1 (SGLT1) and fatty acid translocase (CD36) in the gastrointestinal tract of Holstein cattle and Saanen goats before and after weaning was investigated. Before weaning, the expression of both SGLT1 and CD36 was highest in the jejunum, relative to the other parts of the gastrointestinal tract in both species. The expression of SGLT1 and CD36 in the duodenum was second highest in the goats. After weaning, SGLT1 and CD36 expression in the small intestine significantly decreased in both species. The expression of both types of transporters was also detected in the forestomach. From these results, it was concluded that the jejunum is probably the major absorption site for glucose and long‐chain fatty acids before weaning, and that the expression of both types of transporters decreases after weaning in cattle and goats.     

<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

Twenty-eight new microsatellite loci in chicken and their cross-species amplification in Japanese quail and helmeted guinea fowl

Twenty‐eight original chicken microsatellite markers were isolated and characterized to determine their utility as cross‐reactive markers for comparative genetic mapping in the order Galliformes. Primer pairs were typed in 12 unrelated chickens and also tested on Japanese quail and helmeted guinea fowl deoxyribonucleic acid (DNA). Polymorphism was observed in 23 (82.1%) of the markers and the average number of alleles per locus was 2.9 while the mean heterozygosity was 0.19. Eleven (39.3%) of the chicken markers cross‐reacted with Japanese quail DNA and 2 (7.1%) with helmeted guinea fowl DNA. The cross‐reactive markers described would serve as useful resources for comparative genetic mapping in poultry species belonging to the order Galliformes.     

Chemical Characteristics in Stemflow of Japanese Cedar in Japan

To clarify the characteristics in stemflow of Japanese cedar (Cryptomeria japonica), we conducted the annual and extensive observation. We examined the chemistry of bulk and wet deposition, throughfall and stemflow at 26 forested sites in June and September 1998. The each sampling site was broadly distributed in all over the Japanese archipelago. The stemflow pH of Japanese cedar was significantly lower (p<0.01) than precipitation and stemflow of broad-leaved species in both months. There were significant anion deficits in stemflow of Japanese cedar, suggesting that organic anions derived from plant sources play an important role in the stemflow acidity. Our results suggested that the strong stemflow acidity in Japanese cedar was derived from an internal biological characteristic rather than influences of external acidic deposition.     

Analysis of Proton Generation and Consumption of Forest Surface Soils in Hokkaido, Northern Japan

We determined proton budgets of surface soils in a deciduous forest (Df) and a coniferous forest (Cf) of Volcanogenous Regosols in Tomakomai, Hokkaido of northern Japan. The total H⁺ source was 12.9 and 11.6 kmol_c ha^?1 y^?1 at Df and Cf respectively, and the external H⁺ was 1% at Df and 2% at Cf. The primary H⁺ sources were vegetation uptake of base cations and nitrification, while the major H⁺ sinks were release of base cations and NO₃ ⁺ uptake by vegetation. Leaching incubation experiments using A horizon soils including Df and Cf with NH₄ ⁺ solutions (5.3, 15.9 mg N L^?1) showed that H⁺ from nitrification was generally higher in the Df soil than Cf soil, and nitrification of Tomakomai Df soil was the highest in both treatments. Results of multiple regression analyses suggested that pH_kCl and exchangeable Ca²⁺ contributed to the H⁺ generation via nitrification. Leaching experiments with dilute HCl (pH 3.3) revealed that cation release (mainly Ca²⁺) occurred, and the proportion of release by decrease of exchangeable cations was higher than that by mineral weathering. Mineral weathering in the Tomakomai soil was higher than the other soils.