Multiple copies of <Emphasis Type="Italic">AMT2</Emphasis> are prerequisite for the apple pathotype of <Emphasis Type="Italic">Alternaria alternata</Emphasis> to produce enough AM-toxin for expressing pathogenicity

The apple pathotype of Alternaria alternata produces the cyclic depsipeptide AM-toxin and causes Alternaria blotch of apple. Previously, we cloned AMT2 from the apple pathotype as an orthologue of AFTS1, which is required for biosynthesis of the decatrienoic acid ester AF-toxin I of the strawberry pathotype. These genes were predicted to encode aldo-keto reductases involved in biosynthesis of a common precursor, 2-hydroxy-isovaleric acid, of AF-toxin I and AM-toxin. In this study, we analyzed the function of AMT2 in AM-toxin biosynthesis in the apple pathotype. DNA gel blot analysis of the apple pathotype strain IFO8984 with five restriction enzymes suggested that this strain has a single copy of AMT2 in the genome. However, gene disruption experiments showed that IFO8984 probably has three copies of AMT2. We made mutants having one or two copies of AMT2 disrupted. The single-copy mutants produced less AM-toxin than did the wild type and were still as pathogenic as the wild type. The two-copy mutants produced trace or undetectable amounts of AM-toxin and were markedly reduced in pathogenicity. Thus, AMT2 was verified to be required for AM-toxin biosynthesis and hence pathogenicity. The fact that the two-copy mutants have a remaining copy of AMT2 suggests that multiple copies of AMT2 are prerequisite for the pathogen to produce enough AM-toxin for full pathogenicity.     

Accumulation and localization of cesium in edible mushroom (Pleurotus ostreatus) mycelia

The characteristics of Cs accumulation and localization in edible mushrooms were examined using the mycelia of Pleurotus ostreatus-Y1. Scanning electron microscope images revealed the existence of white spots, and energy dispersive X-ray microanalyzer analysis indicated the presence of larger amounts of Cs and P in these spots in mycelia cultured on medium containing 25 mM CsCl. The (137)Cs activities in the mycelia were approximately 4-6 times higher than those in water used for (137)Cs elution. Higher Cs concentrations in the sediment fraction including vacuolar pellets were obtained compared to the upper fractions. It was observed that yellowish spots caused by the fluorescence of 4',6-diamidino-2-phenylindole (DAPI)-stained polyphosphate were localized in the mycelia. The higher fluorescence intensity of the yellowish-grained spots was measured in comparison with other regions in the mycelium. These results suggested that Cs in the mycelia was trapped by polyphosphate in vacuoles or other organelles.     

Detection of Brucella canis and Leptospira interrogans in canine semen by multiplex nested PCR

Brucella canis and Leptospira interrogans are pathogenic bacteria that cause brucellosis and leptospirosis in dogs around the world. Both diseases can be diagnosed serologically, but the direct detection of these organisms in canine semen is needed when it is used for artificial reproduction. We have been attempting the artificial reproduction of guide dogs for greater breeding efficiency and for this purpose have developed a multiplex nested PCR technique for the detection of B. canis and L. interrogans in the semen and cryoprotective agent (CPA). Our results demonstrated the high sensitivity and simplicity of this technique in the detection of these organisms in canine semen and that will be useful in routine diagnosis. Since they have been found to stay alive in canine semen and CPA up to 48 hr, canine semen for breeding purposes should be checked for contamination by the PCR assay.     

Genetic Analyses of an H3N8 Influenza Virus Isolate, Causative Strain of the Outbreak of Equine Influenza at the Kanazawa Racecourse in Japan in 2007

In August 2007, an outbreak of equine influenza occurred among vaccinated racehorses with Japanese commercial equine influenza vaccine at Kanazawa Racecourse in Ishikawa prefecture in Japan. Apparent symptoms were pyrexia (38.2-41.0 degrees C) and nasal discharge with or without coughing, although approximately half of the infected horses were subclinical. All horses had been shot with a vaccine that contained two inactivated H3N8 influenza virus strains [A/equine/La Plata/93 (La Plata/93) of American lineage and A/equine/Avesta/93 (Avesta/93) of European lineage] and an H7N7 strain (A/equine/Newmarket/1/77). Influenza virus, A/equine/Kanazawa/1/2007 (H3N8) (Kanazawa/07), was isolated from one of the nasal swab samples of diseased horses. Phylogenetic analysis indicated that Kanazawa/07 was classified into the American sublineage Florida. In addition, four amino acid substitutions were found in the antigenic sites B and E in the HA1 subunit protein of Kanazawa/07 in comparison with that of La Plata/93. Hemagglutination-inhibition (HI) test using 16 serum samples from recovering horses revealed that 1.4- to 8-fold difference in titers between Kanazawa/07 and either of the vaccine strains. The present findings suggest that Japanese commercial inactivated vaccine contributed to reducing the morbidity rate and manifestation of the clinical signs of horses infected with Kanazawa/07 that may be antigenically different from the vaccine strains.     

Genetic structure and relationships of 16 Asian and European cattle populations using DigiTag2 assay

In this study, we genotyped 117 autosomal single nucleotide polymorphisms using a DigiTag2 assay to assess the genetic diversity, structure and relationships of 16 Eurasian cattle populations, including nine cattle breeds and seven native cattle. Phylogenetic and principal component analyses showed that Bos taurus and Bos indicus populations were clearly distinguished, whereas Japanese Shorthorn and Japanese Polled clustered with European populations. Furthermore, STRUCTURE analysis demonstrated the distinct separation between Bos taurus and Bos indicus (K=2), and between European and Asian populations (K=3). In addition, Japanese Holstein exhibited an admixture pattern with Asian and European cattle (K=3‐5). Mongolian (K=13‐16) and Japanese Black (K=14‐16) populations exhibited admixture patterns with different ancestries. Bos indicus populations exhibited a uniform genetic structure at K=2‐11, thereby suggesting that there are close genetic relationships among Bos indicus populations. However, the Bhutan and Bangladesh populations formed a cluster distinct from the other Bos indicus populations at K=12‐16. In conclusion, our study could sufficiently explain the genetic construction of Asian cattle populations, including: (i) the close genetic relationships among Bos indicus populations; (ii) the genetic influences of European breeds on Japanese breeds; (iii) the genetic admixture in Japanese Holstein, Mongolian and Japanese Black cattle; and (iv) the genetic subpopulations in Southeast Asia.     

Visualization of musculature of the larval greater amberjack Seriola dumerili (Risso) using whole‐mount immunostaining with special reference to cranial muscles

Muscles play important roles in feeding, respiration and swimming during not only adult stage but also larval stage in fish. However, there is no information of the muscle development in larval greater amberjack, Seriola dumerili (Risso). Here, we investigated muscle development in the greater amberjack, focusing primarily on the cranial muscles from 0 days post hatch (dph) to 12 dph using a modified whole‐mount immunohistochemical staining method. We found that the muscles required for feeding develop by 3 dph, when the larvae begin to feed. Subsequently, muscle composition in the dorsal branchial arches changes to the adult form between 5 and 8 dph. At 8 dph, all the muscles required for feeding and respiration appear, whereas the dorsal, pelvic and caudal fin muscles required for swimming develop later. This report provides fundamental information on larval greater amberjack muscle development, which will enable the detection of abnormal larval muscles and improve larval rearing techniques by modifying environmental conditions.     

Evaluation of chloroform formed in process of kraft pulp bleaching mill using chlorine dioxide

Chloroform formation, balance, and discharge were estimated in a kraft pulp mill that used an elemental chlorine-free (ECF) bleaching line with chlorine dioxide as well as an effluent treatment process. This was achieved by collecting and analyzing discharge water and air samples that contained measurable levels of chloroform. Chloroform formation in ECF bleaching of hardwood oxygen-delignified kraft pulp (LOKP) was estimated to be 2.07–5.34g/pulp air-dried ton (adt), and approximately 30% of the chloroform produced was discharged to bleaching effluents. Chloroform in the effluents was not decomposed by activated sludge, and more than 97% was emitted to the air by volatilization. It is suggested that chloroform formation in Japanese LOKP bleaching mills can be decreased to 16–42t/year with a production of 8 million adt pulp by introducing ECF bleaching into all mills. This is a considerable fall from the chloroform discharge from ECF and chlorine bleaching mills in Japan that was estimated to be approximately 1000t for the 1999 fiscal year.     

日本的奶业政策连载一

边境措施,如关税和关税配额(tariff-rate quotas,TRQs)为日本加工用牛奶生产者提供了强大的政策支持,并使乳制品在日本国内的消费价格高于世界水平.由于与日本进行饮用奶贸易非常困难,因此日本的饮用奶市场在很大程度上是独立于世界市场之外的.日本奶牛养殖场牛奶生产成本较高,再加上相对高的加工与配送成本,日本饮用奶的消费价格也较高.日本采用生产配额制来指导生奶的生产量,以此限制牛奶的供给,并保持市场价格的稳定.政府预算根据奶农所生产的、用于加工的牛奶的配额,每公斤向奶农提供直接、固定的补贴.当加工用牛奶的价格下降时,由奶牛养殖场共享的基金和政府提供的捐助,会根据牛奶的现价与历史平均价格的差价向奶农提供部分补助.日本政府对大部分关税配额内的贸易进行控制,并且对关税配额外、由私人贸易者进口的乳制品征收极高的关税.最近有关乳制品商标标识管理办法的改变,使对用奶粉生产的复原乳的需求明显下降,而对液态奶的需求则出现上升.     

Quantitative trait loci for whiteness of cooked rice detected in improved rice cultivars in Hokkaido

Improving the eating quality of cooked rice has been one of the most important objectives in rice breeding programs. Eating quality of cooked rice is a complex trait including several components, such as external appearance, taste, aroma, and texture. Therefore, dissection of these components followed by marker-assisted selection of detected QTL(s) may be a useful approach for achieving desirable eating quality in rice breeding. Whiteness of cooked rice (WCR) is an important factor related to the external appearance of cooked rice. WCR is known to be associated with the amylose and protein contents of the endosperm. However, the genetic basis of WCR remains unclear. In this study, we evaluated phenotypic variation in WCR among recently developed rice cultivars from Hokkaido, Japan. Then, we developed doubled haploid lines (DHLs) derived from a cross between two cultivars from Hokkaido, Joiku No. 462 (high WCR) and Jokei06214 (low WCR). Using the DHLs, we detected two QTLs for WCR, qWCR3 and qWCR11, on chromosomes 3 and 11, respectively. We also examined the dosage effect of the two QTLs based on both the categorized segregants in the DHLs and the relationship between the WCR phenotype and inheritance around the QTL regions in cultivars from Hokkaido.     

Characteristics of Nitrogenous Air Pollutants at Urban and Suburban Forested Sites, Western Japan

Nitrogenous air pollutants including nitrogen dioxide (NO₂), nitric acid (HNO₃), nitrate (NO ₃ ^? ), ammonia (NH₃), ammonium (NH ₄ ⁺ ), and nitrous acid (HONO) were characterized at an urban forested (UF) site in Hiroshima and at a suburban forested (SF) site in Fukuoka, western Japan, using an annular denuder system for 1?year from May 2006 to May 2007 to compare the concentrations and chemical species of atmospheric nitrogenous pollutants between UF and SF sites. The proximity of the urban area was reflected in higher NO₂ concentrations at the UF site than at the SF site. NO₂ was more oxidized at the SF site because it is farther from an urban area than the UF site, which was reflected in higher concentrations of HNO₃ at the SF site than the UF site. HNO₃ and acidic sulfate is neutralized by NH₃, existing as ammonium nitrate (NH₄NO₃) and ammonium sulfate [(NH₄)₂SO₄] at the UF site. At the SF site, acidic sulfate is neutralized by NH₃, existing as (NH₄)₂SO₄, but NH₄NO₃, had scarcely formed at the SF site. A much higher HONO concentration was observed at the UF site than at the SF site, especially in winter and spring at night, which could be explained by higher NO₂ concentrations at the UF site because of its proximity to an urban area and stagnant meteorological conditions. Atmospheric HONO determination was critical in evaluating the possibility of damage to trees in UF areas.