Phylogenetic Relationships Among Microsporidia Based on rDNA Sequence Data, With Particular Reference to Fish-Infecting Microsporidium Balbiani 1884 Species.

Recently, large discrepancies have been identified between microsporidian systematics based on molecular and traditional characteristics. In the current study the 530f-580r region of the rRNA gene of eight microsporidian species was cloned and sequenced. Included were two unclassified species of Microsporidium Balbiani, 1884 and an unidentified microsporidian that infects the musculature of different sea bream species. Sequence identities in excess of 98% indicated that these three species almost certainly are members of the same genus. Phylogenetic analyses of all microsporidian sequence data available for this region of the gene (20 species) and for partial small subunit sequences (51 species of 21 genera) revealed these species to be distinct from the family Pleistophoridae Doflein, 1901 and closely related them to the genus Sproguea Weissenberg, 1976. This clade was found to comprise a sister taxon to that containing the vast majority of fish-infecting species. Broad cladistic divisions were found between terrestrial insect-infecting and fish-infecting species, which together are distant from the aquatic insect-infecting microsporidia. The rRNA gene of certain fish-infecting genera was found to be more highly conserved than previously reported. This has implications for its utility in diagnostic assays and phylogenetic studies at, or close to, the species level.     

Effects of elevated CO2 on the size structure in even-aged monospecific stands of Chenopodium album

To investigate the effect of elevated CO₂ on the size inequality and size structure, even‐aged monospecific stands of an annual, Chenopodium album, were established at ambient and doubled CO₂ with high and low nutrient availabilities in open top chambers. The growth of individual plants was monitored non‐destructively every week until flowering. Elevated CO₂ significantly enhanced plant growth at high nutrients, but did not at low nutrients. The size inequality expressed as the coefficient of variation tended to increase at elevated CO₂. Size structure of the stands was analyzed by the cumulative frequency distribution of plant size. At early stages of plant growth, CO₂ elevation benefited all individuals and shifted the whole size distribution of the stand to large size classes. At later stages, dominant individuals were still larger at elevated than at ambient CO₂, but the difference in small subordinate individuals between two CO₂ levels became smaller. Although these tendencies were found at both nutrient availabilities, difference in size distribution between CO₂ levels was larger at high nutrients. The CO₂ elevation did not significantly enhance the growth rate as a function of plant size except for the high nutrient stand at the earliest stage, indicating that the higher biomass at elevated CO₂ at later stages in the high nutrient stand was caused by the larger size of individuals at the earliest stage. Thus the effect of elevated CO₂ on stand structure and size inequality strongly depended on the growth stage and nutrient availabilities.     

Proliferation and Neoplastic Transformation of Pigment Cells in Metallothionein/ret Transgenic Mice

Although melanoma is a common human disease, there were few animal models in which melanoma developed at high incidence. To date, the Xiphophorus fish has been used as a model system to study melanoma formation. Studies on this fish showed the presence of a dominant oncogene, Tu, which encodes a transmembrane, tyrosine kinase of epidermal growth factor receptor type (Wittbrodt et al., Nature, 341:415–421, 1989). Recently, we succeeded in establishing novel transgenic mouse lines in which melanosis and melanocytic tumors developed stepwise by introducing another transmembrane tyrosine kinase oncogene, ret (Iwamoto et al., EMBO J., 10:3167–3175, 1991). In our transgenic mice, high levels of expression of the ret transgene induced proliferation and neoplastic transformation of melanin-producing cells. In addition, crossbreeding experiments between transgenic mice and W^v mice showed that the ret oncogene can also induce melanogenesis and melanocyte development in W^v/W^v mice.     

Six polymorphic microsatellite loci in the Japanese Spanish mackerel,Scomberomorus niphonius

Six polymorphic microsatellite loci were isolated for genetic structure studies of the Japanese Spanish mackerel, Scomberomorus niphonius, a severely exploited marine fish species. The number of observed alleles for each locus ranged from seven to 41, and the values of expected and observed heterozygosities were 0.546–0.946 and 0.602–1.000, respectively. These microsatellite markers should prove to be a useful tool for estimating the population genetic structure and genetic variability of Japanese Spanish mackerel.     

DEFORMATION MOVEMENT INDUCED BY DIVALENT IONOPHORE A23187 IN THE TUBIFEX EGG

Deformation movement (groove-forming activity of the equatorial surface) was induced in the spherical Tubifex egg by treatment with ionophore A23187 (5–10 μg/ml). The degree of induced movement was maximal when the treatment was commenced at the beginning of the round stages, i.e. , immediately after disappearance of the grooves formed in the preceding normal deformation movement; it gradually decreased as the onset of the treatment was delayed. This induction was independent of either external Ca²⁺ or Mg²+. Ultrastructural examination of eggs undergoing the induced movement revealed the presence of organized microfilaments in the cortical layer of the grooving regions. It is assumed that the absence of contractile activity during the round stages of the Tubifex egg is attributed basically to the low availability of intracellularly stored Ca²⁺. The results also suggest that the distribution and/or the amount of the contractile elements change during these stages.     

Assessment of the methane mitigation potentials of alternative water regimes in rice fields using a process‐based biogeochemistry model

Rice production is a substantial source of atmospheric CH₄, which is second only to CO₂ as a contributor to global warming. Since CH₄ is produced in anaerobic soil environments, water management is expected to be a practical measure to mitigate CH₄ emissions. In this study, we used a process‐based biogeochemistry model (DNDC‐Rice) to assess the CH₄ mitigation potentials of alternative water regimes (AWR) for rice fields at a regional scale. Before regional application, we tested DNDC‐Rice using site‐scale data from three rice fields in Japan with different water regimes. The observed CH₄ emissions were reduced by drainage of the fields, but were enhanced by organic amendments. DNDC‐Rice gave acceptable predictions of variation in daily CH₄ fluxes and seasonal CH₄ emissions due to changes in the water regime. For regional application, we constructed a GIS database at a 1 × 1 km mesh scale that contained data on rice field area, soil properties, daily weather, and farming management of each cell in the mesh, covering 3.2% of the rice fields in Japan's Hokkaido region. We ran DNDC‐Rice to simulate CH₄ emissions under five simulated water regimes: the conventional water regime and four AWR scenarios with gradually increasing drainage. We found that AWR can reduce CH₄ emission by up to 41% compared with the emission under conventional water regime. Including the changes in CO₂ and nitrous oxide emissions, potential mitigation of greenhouse gas (GHG) was 2.6 Mg CO₂ Eq. ha^?1 yr^?1. If this estimate is expanded to Japan's total rice fields, expected GHG mitigation is 4.3 Tg CO₂ Eq. yr^?1, which accounts for 0.32% of total GHG emissions from Japan. For a reliable national‐scale assessment, however, databases on soil, weather, and farming management must be constructed at a national scale, as these factors are widely variable between regions in Japan.     

Hot spots of leaf breakdown within a headwater stream reach: comparing breakdown rates among litter patch types with different macroinvertebrate assemblages

1. Detecting hot spots of litter decomposition will promote understanding of litter processing in a heterogeneous system. To identify hot spots of leaf breakdown within a headwater stream reach, we examined the difference in leaf breakdown rate among four types of litter patches, one that formed in riffles and three that formed in pools (middle, alcove, edge), in different seasons. 2. Middle patches showed the highest breakdown rate in some seasons; the rate in middle patches was 1.5–4 times higher than in the other patches. Thus middle patches can be regarded as hot spots of leaf breakdown in the study reach. This result contrasted with other studies showing higher breakdown rate in riffles than in pools. 3. Significant relationships between abundance of caddisfly shredders and breakdown rate were observed in seasons when the rate differed among patch types. Greater abundance of Lepidostoma seems to be responsible for middle patches being hot spots of leaf breakdown. 4. It is expected that when the proportion of leaves retained in middle patches within a reach is higher, the breakdown rate of the entire reach will be increased. Clarifying how the proportion of leaves retained on middle patches within a reach varies temporally and spatially would improve our understanding of leaf breakdown in headwater streams.     

Morphological responses of Daphnia ambigua to different concentrations of a chemical extract from Chaoborus flavicans

1. Daphnia ambigua were reared individually with eight different concentrations of a chemical extract from larvae of Chaohorus flavicans, and the frequency of appearance of helmeted individuals and the helmet sizes were determined in the instars 1–6. 2. The animals developed most marked helmets in the second instar in response to the extract. The frequency of the helmeted individuals and helmet size in the second instar increased with increasing chemical concentration, although their dose-response curves differed between them. 3. The results may explain the cyclomorphosis of Daphnia, which develop the highest helmets in summer, when the predators are abundant and active, and the concentrations of the predator-released chemicals would presumably be high. 4. The extract became toxic to Daphnia at the highest concentration prepared.     

Maternal and paternal effects on the germination time of non‐dormant seeds of a monocarpic perennial species,Aster kantoensis (Compositae)

In plant species producing non‐dormant seeds, the germination time (from the start of imbibition to radicle emergence) is the main factor determining the timing of seedling emergence. We investigated maternal and paternal genetic effects on the germination time of non‐dormant seeds of a monocarpic perennial, Aster kantoensis Kitamura (Compositae). Three sets of reciprocal diallel crosses among five plants were conducted to produce genetic variation in seeds, and the germination time of the progeny of each parent was determined. The effects of the maternal parent and the interaction of maternal and paternal parents on the germination time of progeny were significant in all sets, and the effect of the paternal parent was significant in two of the three sets. This result means that the germination time of the progeny of a maternal or paternal parent can vary with the genotype of its mating partners. Because variation in the emergence time of seedlings contributes to avoiding seedling loss owing to unpredictable environmental changes, genetic variation in the germination time among the progeny of each parent mating with multiple partners could contribute to the establishment of the parent's seedlings in species producing non‐dormant seeds in the field.     

基于线粒体DNA序列探讨斑头鱼分类地位

测定了斑头鱼Agrammus agrammus和大泷六线鱼Hexagrammos otakii的线粒体COⅠ、Cyt b和16S rRNA基因的部分序列,结合从GenBank中获得的六线鱼属3种的同源序列,以单鳍多线鱼Pleurogrammus monopterygius为外群,运用邻接法(NJ)、最大简约法(MP)和贝叶斯法(BI)构建了分子系统树。同时联合了3个基因片段序列,运用贝叶斯联合模型综合探讨了六线鱼类的系统发育关系。结果显示:除16S rRNA基因外,其余2个基因片段以及联合模型所构建的系统树拓扑结构完全一致,即斑头鱼与大泷六线鱼亲缘关系最近,应归为六线鱼属,拉丁学名应为Hexagrammos agrammus。Cyt b基因片段序列分析结果显示,斑头鱼和大泷六线鱼分歧时间约为175万年。结合形态学研究资料,支持将斑头鱼归为六线鱼属的观点,斑头鱼和大泷六线鱼亲缘关系最近,属于六线鱼科中分化较晚的种类。