A phylogeographic study of the Japanese earthworm,Metaphire sieboldi (Horst, 1883) (Oligochaeta: Megascolecidae): Inferences from mitochondrial DNA sequences

To clarify the historical phylogeography of a Japanese earthworm, nucleotide sequence variations of Metaphaire sieboldi were investigated with respect to its geographic distribution. Sequencing of mitochondrial DNA (mtDNA), including the cytochrome oxidase subunit I (COI) region and the 16S rDNA region, of 72 samples including 4 outgroup taxa, and phylogenetic analyses using neighbour-joining (NJ) and Bayesian inference (BI) methods indicated that M. sieboldi forms a monophyletic group. This monophyletic group was divided into seven subgroups; most subgroups comprised individuals from several areas, except for southern Shikoku and southern Kyushu. The phylogenetic tree demonstrated that the Shikoku and Kinki populations were paraphyletic at the basal node, and individuals from Kyushu and western Shikoku were positioned at the advanced group. Estimated phylogeographic events based on mtDNA analyses include (1) M. sieboldi originated in Shikoku and/or Kinki, (2) individuals from Chugoku and Kyushu have a common ancestor from Shikoku, and (3) individuals from western Shikoku originated from Kyushu progenitors (about 0.4–0.5 mya) after separation by rising sea levels.     

Mapping of QTLs controlling epicotyl length in adzuki bean (Vigna angularis)

Epicotyl length (ECL) of adzuki bean (Vigna angularis) affects the efficiency of mechanized weeding and harvest. The present study investigated the genetic factors controlling ECL. An F₂ population derived from a cross between the breeding line ‘Tokei1121’ (T1121, long epicotyls) and the cultivar ‘Erimo167’ (common epicotyls) was phenotyped for ECL and genotyped using simple sequence repeats (SSRs) and single-nucleotide polymorphism (SNP) markers. A molecular linkage map was generated and fifty-two segregating markers, including 27 SSRs and 25 SNPs, were located on seven linkage groups (LGs) at a LOD threshold value of 3.0. Four quantitative trait loci (QTLs) for ECL, with LOD scores of 4.0, 3.4, 4.8 and 6.4, were identified on LGs 2, 4, 7 and 10, respectively; together, these four QTLs accounted for 49.3% of the phenotypic variance. The segregation patterns observed in F₅ residual heterozygous lines at qECL10 revealed that a single recessive gene derived from T1121 contributed to the longer ECL phenotype. Using five insertion and deletion markers, this gene was fine mapped to a ~255 kb region near the end of LG10. These findings will facilitate marker-assisted selection for breeding in the adzuki bean and contribute to an understanding of the mechanisms associated with epicotyl elongation.     

An institutional case study of Japanese Water Users Association: towards successful participatory irrigation management

This paper examines irrigation management conduct within the Japanese Water Users Association (WUA), namely the Land Improvement District (LID). LID is regarded as a successful case of participatory irrigation management (PIM), in which there are few conflicts over water between farmers so that it attains high social cost performance. However, the management system of LID still remains obscure. We throw light on this system assuming the existence of "the rule of fairness". Farmers operate the irrigation facilities in a self-serving way and have their own rule of fairness, such as upstream superiority, based on the long-term experiences of irrigation systems. We defined this rule of fairness as made up of two components: one is "fairness of outcome" and the other is "fairness of procedure". Finally, we discovered that the approach of LID staff is the essential factor to satisfying the farmers' criteria of fairness and maintaining peace; LID staff place importance on farmers' customary rules, taking care to remain neutral in arbitrating quarrels between farmers and trying to ensure organizational transparency to farmers. This can be also a good lesson for future PIM projects.     

Uptake by dietary exposure and elimination of aflatoxins in muscle and liver of rainbow trout (Oncorhynchus mykiss)

Uptake and elimination of aflatoxins (AFs) by rainbow trout ( Oncorhynchus mykiss ) during a long-term (21 days) dietary exposure were studied to assess contamination by AFs in aquaculture fish fed AF-containing feed. The uptake factor (UF) of aflatoxin B(1) (AFB(1)) in muscle ranged from 0.40 × 10(-3) to 1.30 × 10(-3). AFB(1) concentrations in liver were 165-342 times higher than in muscle. AFs from feed were more highly accumulated in liver than in muscle. Aflatoxicol (AFL) and aflatoxin M(1) (AFM(1)) were detected in muscle and liver and also in the rearing water. AFL concentrations were higher than AFM(1) by 2 orders of magnitude in muscle, and AFL was a major metabolite of AFB(1). The elimination rate constants (α) of AFB(1) and AFL in muscle (1.83 and 2.02 day(-1), respectively) and liver (1.38 and 2.41 day(-1), respectively) were very large. The elimination half-life (t(1/2)) of AFB(1) was 0.38 days (9.12 h) in muscle and 0.50 days (12.00 h) in liver. The elimination half-life of AFL in muscle and liver was 0.34 day (8.16 h) and 0.29 day (6.96 h), respectively. These data show that AFs are eliminated rapidly and are not biomagnified in fish. Thus, AFB(1) concentration in muscle of fish fed AFB(1)-containing feed (ca. 500 μg/kg) decreased to below the detection limit (20 ng/kg) of the most sensitive analytical method at 1.54 days (36.96 h) after the change to uncontaminated feed.     

Triterpenoids isolated from Zizyphus jujuba inhibit foam cell formation in macrophages

Because foam cell formation in macrophages is believed to play an essential role in the progression of early atherosclerotic lesions in vivo, prevention of foam cell formation is considered to be one of the major targets for the treatment of atherosclerosis. The present study examined the inhibitory effect of 50 crude plant extracts on foam cell formation. Among those crude extracts, Zizyphi Fructus (ZF) and Zizyphi Semen (ZS) extracts significantly inhibited the foam cell formation induced by acetylated LDL. Furthermore, triterpenoids such as oleanonic acid, pomolic acid, and pomonic acid were the major active compounds, and triterpenoids containing a carboxylic acid at C-28 play an important role in the inhibitory effect on foam cell formation in human macrophages. These data suggest that triterpenoids in Zizyphus jujuba , the plant source of ZF and ZS, may therefore be useful for the prevention of atherosclerosis.     

Effects of land-use type and nitrogen addition on nitrous oxide and carbon dioxide production potentials in Japanese Andosols

Land-use type and nitrogen (N) addition strongly affect nitrous oxide (N₂O) and carbon dioxide (CO₂) production, but the impacts of their interaction and the controlling factors remain unclear. The aim of this study was to evaluate the effect of both factors simultaneously on N₂O and CO₂ production and associated soil chemical and biological properties. Surface soils (0–10 cm) from three adjacent lands (apple orchard, grassland and deciduous forest) in central Japan were selected and incubated aerobically for 12 weeks with addition of 0, 30 or 150 kg N ha^–1 yr^–1. Land-use type had a significant (p < 0.001) impact on the cumulative N₂O and CO₂ production. Soils from the apple orchard had higher N₂O and CO₂ production potentials than those from the grassland and forest soils. Soil net N mineralization rate had a positive correlation with both soil N₂O and CO₂ production rates. Furthermore, the N₂O production rate was positively correlated with the CO₂ production rate. In the soils with no N addition, the dominant soil properties influencing N₂O production were found to be the ammonium-N content and the ratio of soil microbial biomass carbon to nitrogen (MBC/MBN), while those for CO₂ production were the content of nitrate-N and soluble organic carbon. N₂O production increased with the increase in added N doses for the three land-use types and depended on the status of the initial soil available N. The effect of N addition on CO₂ production varied with land use type; with the increase of N addition doses, it decreased for the apple orchard and forest soils but increased for the grassland soils. This difference might be due to the differences in microbial flora as indicated by the MBC/MBN ratio. Soil N mineralization was the major process controlling N₂O and CO₂ production in the examined soils under aerobic incubation conditions.     

Characteristics and classification of an unirrigated anthropogenic-alluvial soil found in the Kumamoto Plain

In Japan anthropogenic-alluvial soils excluding irrigated rice soils are lagging for behind other genetic soil types in pedological investigations. Although there are some investigations of anthropogenic-alluvial soils used for mulberry plantations, orchards, vegetable gardens, and seeding beds for trees, the main object of these investigations is not pedological. Kanno et al. (1) reported that unirrigated anthropogenic-alluvial soils used for vegetable growing in the Chikugo Plain resemble the “Alochthonous brown warp soil” defined by Kubiena (2). First of all, it is of great importance to accumulate general data for unirrigated anthropogenic-alluvial soils and to develop the classification principles for them. Classification problems will be settled in accordance with the accumulated data. This paper deals with characteristics of an unirrigated anthorpogenic-alluvial soil found in the Kumamoto Plain and with classification principles for it. These principles have almost never been described for a pedological standpoint in Japan.     

Allantoin production and its utilization in relation to nodule formation in soybeans

A remarkable accumulation and utilization of allantoin are observed in soybean plant bearing modules (Glycine max variety A62-1). To study ita physiological role, changes in activities and distribution of the enzymes concerning the purine catabolism (i.e., xanthine oxidase, uricase, allantoinase and allantoicase) were measured during development.

Uricase activity was found in the radicles in the early stage of seedling development, and no difference of the enzyme activity was detected between the radicles of the nodulating variety (A62-) and non-nodulating variety (A62-2). On maturation the activity disappeared rapidly in radicles and appeared in leaves, pods and nodules. The activity in nodules was much higher than that in the leaves and the pods throughout the culture period.

Xanthine oxidase activity was found in the radicles and nodules.

Allantoinase activity was present in all regions of both varieties, and the specific activity in the nodules Was 2-10 times that in other regions.

Allantoicase activity was detected in the leaves, but not in the nodules.

The distribution of these enzymes suggested that allantoin is formed in the nodules and utilized mainly in the leaves after translocation.

Changes in the allantoin concentration and in the activities of uricase and allantoicase were studied in non-nodulating soybean (A62-2) grown in a culture solution containing various nitrogenous compounds. The allantoin concentration in the leaves increased as the plants were cultured in the solution containing 100 ppm of nitrate-N, urea-N or allantoin-N, whereas no uricase activity in the root was detected. The specific activity of uricase decreased in the process of plant development in all cultures. Allantoicase activity increased in the roots and leaves when the plants were cultured in this solution containing 100 ppm of urea-N or allantoin-N, indicating that allantoin is utilized easily in the soybean plants.

Intracellular localization of uricase in the nodules was studied by the method of linear sucrose density gradient centrifugation. The uricase activity was associated with the bacteroids, density 1.25.

The uricase activity peak was coincident with the peak of fonnic dehydrogenase activity (bacteria type), and not with peaks of catalase activity, cytochrome oxidase activity and absorbance at 280 nm. This evidence together with the observation by electron microscopy suggested that at uricase is associated with bacteroids.     

Cytological characteristics of microconidia of <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

The inner cellular structure of microconidia of Magnaporthe oryzae was examined using fluorescent probes and electron microscopic techniques. The volume of the nucleus relative to the cell was significantly larger in microconidia than in macroconidia or vegetative hyphae, similar to observations for spermatia of other fungi. Selective fluorescent staining revealed that cytosolic RNA was less abundant in microconidia than in macroconidia and germ tubes, suggesting that general metabolic activity in microconidia is low. Consistently, GFP expression driven by the TrpC promoter was highly active during the formation of phialides and microconidia but gradually decreased as the microconidia matured. Such data suggest that microconidia are in a quiescent or dormant state.     

Morphological and molecular characterization of <Emphasis Type="Italic">Oidium</Emphasis> subgenus <Emphasis Type="Italic">Reticuloidium</Emphasis> (powdery mildew) newly occurred on cucumber in Japan

In 2002, a powdery mildew with catenate conidia lacking fibrosin bodies was found on cucumber in a greenhouse in Kanagawa Prefecture, Japan. Morphological observation revealed that the fungus belongs to Oidium subgenus Reticuloidium, anamorph of the genus Golovinomyces. Molecular phylogenetic analyses of the nucleotide sequences of the rDNA ITS regions and D1/D2 domains of the 28S rDNA indicated that the fungus belongs to the clade of G. orontii with other Golovinomyces fungi from a wide range of host plants, suggesting that the fungus was newly transported from abroad. Because there has been no prior report of cucumber powdery mildew caused by Reticuloidium, further research on the physiology, epidemiology, control and resistant cucumber varieties is required.