Two phases of intracellular reactive oxygen species production during victorin-induced cell death in oats

Reactive oxygen species (ROS) are thought to be involved in various forms of programmed cell death (PCD) in animal and plant cells. PCD, along with the production of ROS, occurs during plant–pathogen interactions. Here we show that victorin, a host-specific toxin produced by Cochliobolus victoriae, which causes victoria blight of oats, induces two phases of intracellular ROS production in victorin-sensitive oat mesophyll cells. The initial production of ROS is restricted at mitochondria and not accompanied with cellular oxidative damage. Later production of ROS is dispersed into cells concomitant with lipid peroxidation, chloroplast dysfunction, and cell death. Superoxide dismutase can clearly suppress the initial ROS production and delay the progression of cell death. These data indicate that the initial ROS production may be involved in the cell death induction process, and the later ROS production may play important roles in events leading to cellular disruption.     

Significance of PWT4–Rwt4 interaction in the species specificity of Avena isolates of Magnaporthe oryzae on wheat

We examined whether PWT4, an avirulence gene of Avena isolates of Magnaporthe oryzae toward wheat, corresponded to Rwt4, a resistance gene identified in wheat cultivar Norin 4, in a one-to-one manner. Twelve wheat cultivars were inoculated with 65X1, an F₁ culture with PWT4 derived from a cross between an Avena isolate (Br58) and a Triticum isolate (Br48). Three wheat cultivars (Norin 26, Shin-chunaga, Cheyenne) were resistant and therefore selected as possible carriers of Rwt4. The three cultivars were then inoculated with a population derived from a backcross of 61M2 carrying PWT4 with Br48 carrying pwt4. Segregation analyses revealed that PWT4 operates against the three cultivars. If PWT4 corresponds to Rwt4 in a one-to-one manner, all three cultivars should carry Rwt4. To test if this is the case, the three cultivars were crossed with Chinese Spring (a noncarrier of Rwt4) and Norin 4. When F₂ seedlings from Chinese Spring × Norin 26, Chinese Spring × Shin-chunaga, and Chinese Spring × Cheyenne were inoculated with 61M2, resistant and susceptible seedlings segregated in a 3 : 1 ratio. On the other hand, crosses between the three cultivars and Norin 4 yielded no susceptible F₂ seedlings. These results indicate that all three cultivars carry Rwt4. Considering all results, we concluded that PWT4 corresponds to Rwt4 in a one-to-one manner. An inoculation test with Chinese Spring–Cheyenne chromosome substitution lines indicated that Rwt4 is located on chromosome 1D.     

Novel polymorphism of the canine dopamine receptor D4 gene intron II region

Various dog breeds are remarkably different from each other not only in their sizes and shapes but also in behavioral traits, suggesting that some of these characteristics are under genetic control. However, little is known about genes related to behavioral traits in canine species. In humans, it has been reported that the dopamine receptor D4 gene (DRD4) includes polymorphism at several regions that relate to personality or psychiatric disorders. In an earlier study by the authors of the present study, the polymorphisms in canine DRD4 exon III and exon I regions were reported. In the present study, a novel polymorphism in canine DRD4 intron II was found based on a 17 base pair insertion/deletion, and the two alleles detected were named P (shorter allele) and Q (longer allele). The allelic distribution in 28 breeds of dog, including a total of 1114 unrelated individuals, were then investigated. Both P and Q alleles were detected in most of the breeds investigated; however, the frequencies of P and Q differed greatly between breeds. With respect to classification based on breed origin, P and Q alleles were frequent in Occidental and Oriental breeds, respectively. Furthermore, two subspecies of wolves, the ancestors of dogs, were analyzed for the comparison of allele frequencies with dogs, and the P allele was predominant in both European and Chinese wolves.     

Propagation of bovine coronavirus in clones of the Caco-2 cell line showing different levels of alkaline phosphatase activity

The aim of the present study was to determine whether bovine coronavirus (BCV) has the ability to initiate infection in a human colon carcinoma cell line, Caco‐2, that has been established to spontaneously differentiate after confluence. When Caco‐2 cells were infected with BCV, a titer of 5.5 × 10⁶ plaque‐forming units (p.f.u.)/mL was found in the culture supernatant at 5 days postinfection. Two clones, Caco‐2/CA1 and Caco‐2/CA2, were then isolated by monitoring alkaline phosphatase (ALP) and cell proliferation activities. The ALP activity level of CA1 cells was significantly higher than that of CA2 cells, while the level of cell proliferation activity of CA1 was significantly lower than that of CA2. When CA1 and CA2 cells were infected with BCV at confluence, virus hemagglutination (HA) was detected in the culture supernatant at 5 days postinfection for CA1 cells and at 8 days postinfection for CA2 cells. Thus, BCV propagation was substantially delayed in CA2 cells, suggesting that a cellular factor(s) that appears at the differentiation stage may control BCV propagation. BCV‐susceptible CA1 and CA2 cells showing different levels of ALP activity would be useful for further experiments to elucidate the mechanism of BCV propagation.     

Disease resistance induced by nonantagonistic endophytic Streptomyces spp. on tissue-cultured seedlings of rhododendron

Of 82 strains of endophytic actinomycetes isolated from rhododendron plants, 12 were not antagonistic against Pestalotiopsis sydowiana, which is the causal agent of Pestalotia disease. Of these 12, MBR-37 and MBR-38 (identified as Streptomyces spp.) grew on IMA-2 medium. Tissue-cultured seedlings of rhododendron treated with these nonantagonistic strains showed less wilting and/or smaller lesions to P. sydowiana, although the degree of resistance was a little lower than that conferred by antagonistic Streptomyces galbus strain R-5. These seedlings accumulated the anthocyanin(s), suggesting that resistance induced by these strains could depend on activated defense responses associated with the phenylpropanoid pathway rather than with antibiosis.     

Volatile organic compounds from wood and their influences on museum artifact materials I. Differences in wood species and analyses of causal substances of deterioration

Wood is generally used as the interior material in museum storage rooms. Recently, however, the effects of volatile organic compounds (VOCs) from wood on artifacts has become a topic of great concern. The VOCs from four species of wood (western red cedar, spruce, kiri, and sugi) and their effects on artifact materials (two types of metal, seven types of pigment) were investigated using a deterioration-accelerating test, gas chromatography-mass spectrometry, X-ray diffraction analysis, and X-ray photoelectron spectroscopy. The results suggested that the influences on artifact materials varied greatly with wood species, and depended on specific components such as hinokitiol or acetic acid rather than the amount of total volatile organic compounds (TVOCs). It is a very serious problem that of the four species of wood, western red cedar (rich in hinokitiol), which has been recommended as an interior material for museum storage rooms, showed the heaviest deterioration on metal samples, and only this type of wood discolored enpaku (white lead) and rokushou (malachite, verdigris). In such storage rooms, museum artifacts should be carefully monitored. When selecting an interior material for a storage room or studying methods of preventing deterioration, it is very important to consider fully the characteristics of wood VOCs, not only the amount of TVOC.Part of this paper was presented at the 24th (Tokyo, June 2002) and 25th (Kyoto, June 2003) Annual Meeting of the Japan Society for the Conservation of Cultural Property     

Fecal progestagen and estrone during pregnancy in a giraffe: a case report

The present study was carried out to measure fecal progestagen and estrone concentrations during pregnancy in a giraffe and examine the possibility of utilizing this assay system for pregnancy diagnosis. Fecal samples were collected from a giraffe during her third and fourth parities and mixed with methanol to prepare a fecal solution. Diluted fecal solution was used for direct enzyme immunoassay for progestagen and estrone. The newborn calf from the third parity was viable, although that from the fourth parity died 5 days after calving. In the third parity, the giraffe's progestagen and estrone concentrations increased transiently from days 30 to 120 of pregnancy. Then, they decreased and remained low until day 330. This was followed by a drastic rise in both concentrations as parturition approached. Parturition caused a reduction in the progestagen and estrone concentrations of the feces. In the fourth parity, the progestagen concentration increased gradually after mating until day 320. This was followed by a reduction in the concentration until parturition. However, the estrone concentration fluctuated, and the duration and extent of the prepartum rise in concentration were shorter and lower than those of the third parity. The hormone dynamics of the third parity suggest the possibility of early pregnancy diagnosis by measuring progestagen or estrone between days 30 and 120 after mating.     

Japanese black cattle with orotic aciduria detected by gas-chromatography/mass-spectrometry

A 4-months-old calf of Japanese black cattle was diagnosed with orotic aciduria by gas-chromatography/mass-spectrometry (GC/MS). Until now orotic aciduria had not been reported in Japanese black cattle. The animal showed repeated diarrhea. The hematocrit was low, and microcytes and acanthocytes were observed in blood smears. The calf had lower serum total protein concentrations with a higher blood ammonia concentration. Needle-shaped crystals of orotic acid were observed in urinary sediments. Sequence homologous analysis with cattle uridine monophosphate synthase DNA indicated silent mutation in the affected calf.     

中国茶树的起源与分布——根据茶叶香气组分中萜烯指数化学分类所作的推论(英文)

茶挥发性成分中的萜烯醇类具有生物合成的遗传特性。根据萜烯指数(即:芳樟醇及其氧化物/(芳樟醇及其氧化物十香叶醇)比值,TI〕所作的化学分类表明,云南省茶的 TI 值接近于1.0,而分布于其他产茶省区的茶树 TI 值均低于上述值,并伴随有由西南向东和东南逐渐下降的趋势。由云南引入的日本山茶,具有与云南相近的 TI 值,而日本当地茶树品种的 TI 值与中国浙江相近似,因而推测中国茶树的原产地为云南,然后沿长江和沿海逐渐向东和南方传播。     

Mode of bleaching phytotoxicity of herbicidal diphenylpyrrolidinones

A mode of action study of herbicidal diphenylpyrrolidinones was carried out through carotenoid analyses in intact Scenedesmus cells and by a cell‐free plant‐type phytoene desaturase assay using Escherichia coli transformants. A series of forty‐eight diphenylpyrrolidinones decreased the carotenoid content of Scenedesmus cells in the light and inhibited phytoene desaturase. The relationship between substituents at various positions and inhibition of phytoene desaturase is discussed. Using very active bleaching diphenylpyrrolidinones, a 10⁻⁵ M concentration affected neither the ζ‐carotene desaturase nor the protoporphyrinogen‐IX oxidase. Although some differences in their inhibitory activity were found between the in vivo and cell‐free assays, it is concluded that the compounds are essentially bleachers affecting carotenoid biosynthesis in plants. Enzyme kinetics studies with recombinant phytoene desaturase revealed a non‐competitive inhibition with respect to the substrate phytoene. A competition against the inhibitor was shown by the cofactor NADP⁺, suggesting an interaction of pyrrolidinones at the cofactor‐binding site of phytoene desaturase. © 2001 Society of Chemical Industry