Seed germination and seedling establishment of the monocarpic dwarf bamboo Sasa veitchii var. hirsuta

In order to determine the influences of environmental factors on seed germination and to identify the effects of germination characteristics on seedling establishment for Sasa veitchii var. hirsuta, germination experiments and a survey on current seedling dynamics were conducted in a natural habitat after mass flowering. The results of the germination experiments revealed that the seeds require a low-temperature environment for germination; those exposed to low-temperature conditions for a time similar to the length of winter (4 months of low-temperature conditions) germinated gradually, while those exposed for longer (>4 months of low-temperature conditions) germinated rapidly. These results were compatible with field observations indicating that seedlings emerged from June to October in the year after mass flowering, and they suggested this germination characteristic (i.e., variation of germination timing due to seed dormancy) may play an ecological function in spreading risk (although it may also increase the potential for seed predation). The results also revealed that stronger light and alternating temperature conditions had no effect on germination, whereas high temperatures and drying had negative effects. The outcomes of the survey on current seedling dynamics revealed that the seedling establishment ratio was high in a forest understory compared to that seen at a site where solar radiation was strong and soil water content was low. This implies that germination characteristics may promote seedling establishment in forest understory.     

Detection of quinolone-resistance genes in Photobacterium damselae subsp. piscicida strains by targeting-induced local lesions in genomes

Quinolone-resistant strains of the fish-pathogenic bacterium, Photobacterium damselae subsp. piscicida are distributed widely in cultured yellowtail, Seriola quinqueradiata (Temminck & Schlegel), in Japan. The quinolone resistance-determining region (QRDR) was amplified with degenerate primers, followed by cassette ligation-mediated PCR. Open reading frames encoding proteins of 875 and 755 amino acid residues were detected in the gyrA and parC genes, respectively. Resistant strains of P. damselae subsp. piscicida carried a point mutation only in the gyrA QRDR leading to a Ser-to-Ile substitution at residue position 83. No amino acid alterations were discovered in the ParC sequence. A mutation in the gyrA gene was also detected in nalidixic acid-resistant mutants of strain SP96002 obtained from agar medium containing increased levels of quinolone. These results suggest that GyrA, as in other Gram-negative bacteria, is a target of quinolone in P. damselae subsp. piscicida. Furthermore, we attempted to detect a point mutation using targeting-induced local lesions in genomes (TILLING), which is a general strategy used for the detection of a variety of induced point mutations and naturally occurring polymorphisms. We developed a new detection method for the rapid and large-scale identification of quinolone-resistant strains of P. damselae subsp. piscicida using TILLING.     

Regulation of mammalian tooth cusp patterning by ectodin

Mammalian tooth crowns have precise functional requirements but cannot be substantially remodeled after eruption. In developing teeth, epithelial signaling centers, the enamel knots, form at future cusp positions and are the first signs of cusp patterns that distinguish species. We report that ectodin, a secreted bone morphogenetic protein (BMP) inhibitor, is expressed as a "negative" image of mouse enamel knots. Furthermore, we show that ectodin-deficient mice have enlarged enamel knots, highly altered cusp patterns, and extra teeth. Unlike in normal teeth, excess BMP accelerates patterning in ectodin-deficient teeth. We propose that ectodin is critical for robust spatial delineation of enamel knots and cusps.     

Expressed sequence tag analysis of phyllosomas and hemocytes of Japanese spiny lobster Panulirus japonicus

Two complementary DNA (cDNA) libraries were constructed from phyllosomas and hemocytes of adult Japanese spiny lobster Panulirus japonicus and a total of 2,673 expressed sequence tags (ESTs) were obtained. After assembly and clustering, 450 and 458 unique sequences were found from the phyllosoma and hemocyte cDNA libraries, respectively. Of these, 114 and 220 ESTs showed significant homologies with known genes in the National Centre for Biotechnology Information (NCBI) database. The remaining sequences were of unknown function. Immune-related genes found in this study include lectin, proteinase inhibitor, prophenoloxidase, heat-shock protein, antimicrobial peptide, and a few putative defense-related proteins.     

Putative virulence-related genes in Vibrio anguillarum identified by random genome sequencing

The genome of Vibrio anguillarum strain H775-3 was partially determined by a random sequencing procedure. A total of 2,300 clones, 2,100 from a plasmid library and 200 from a cosmid library, were sequenced and subjected to homology search by the BLAST algorithm. The total length of the sequenced clones is 1.5 Mbp. The nucleotide sequences were classified into 17 broad functional categories. Forty putative virulence-related genes were identified, 36 of which are novel in V. anguillarum, including a repeat in toxin gene cluster, haemolysin genes, enterobactin gene, protease genes, lipopolysaccharide biosynthesis genes, capsule biosynthesis gene, flagellar genes and pilus genes.     

Inhibition of hirame rhabdovirus growth by RNA aptamers

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)‐RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV‐RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.     

Availability of genetically modified feed ingredient: investigations of ingested foreign DNA in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

ABSTRACT:   Foreign DNA fragments from genetically modified defatted soybean meal (GM SBM) in rainbow trout was traced by nested polymerase chain reaction (PCR) and located by in situ hybridization. Either a GM or non-GM SBM formulated diet (42% protein) was fed to fish (average weight 50.5 g) for 2 weeks. The degradation results showed that the cauliflower mosaic virus 35S promoter (220 bp) fragment was detected in the contents of digestive system only in fish fed the GM SBM diet, and it was not detected on the third day after changing the diet to the non-GM SBM diet. For the possible transferal results, the promoter fragment was detected in the leukocyte, head kidney and muscle only of fish fed the GM SBM diet; it was not detected on the fifth day after changing the diet to the non-GM SBM diet. These results suggest that a foreign DNA fragment was not completely degraded and might be taken up into organs through the gastrointestinal tract. However, foreign DNA was not detected after the withdrawal period. Thus, the data show that uptake of DNA from GM SBM might not remain in the tissues of fish fed GM SBM diet.     

Quantitative in vivo measurement of central benzodiazepine receptors in the brain of cats by use of positron-emission tomography and [11C]flumazenil

OBJECTIVE: To map central benzodiazepine receptors (BZRs) in the brain of cats by use of positron-emission tomography (PET) and [11C]flumazenil. ANIMALS: 6 male cats that weighed between 2.0 and 3.6 kg. PROCEDURE: Brain images obtained by PET evaluation of [11C]flumazenil were superimposed on T2-weighted magnetic-resonance imaging (MRI) scans of the same cats. Detailed anatomic regions, such as the cerebral cortex, striatum, thalamus, midbrain, and cerebellum, on the PET images were evident by PET-MRI registration. Regional binding of [11C]flumazenil to BZRs was quantitatively measured by use of a model with 2 tissue compartments and 4 variables. RESULTS: The highest value for distribution volume was observed in the cerebral cortex, and the lowest value was found in the midbrain of cats. CONCLUSIONS AND CLINICAL RELEVANCE: Binding of [11C]flumazenil to BZRs in the brain of cats can be quantitatively measured by use of PET with the aid of PET-MRI registration. It is difficult to diagnose changes in these neuroreceptors within the field of current veterinary science. In the future, PET should prove useful for investigating and diagnosing brain disorders in animals in clinical settings.