Chromatographic and immunological identification of gonadotropin-releasing hormone in five marine teleosts

Brain extracts from bluefin tuna, Thunnus thynnus, red seabream, Pagrus major, black seabream, Acanthopagrus schlegeli, red spotted grouper, Epinephelus akaara and Japanese flounder, Paralichthys olivaceus, were analyzed by high performance liquid chromatography (HPLC) and specific radioimmunoassays. Immunoreactive material co-eluting from HPLC with salmon gonadotropin-releasing hormone (GnRH) and chicken GnRH-II, respectively, was found in all five species. In addition, a GnRH immunoreactive fraction showing the same HPLC retention time as lamprey GnRH-I was detected in the brain extracts of all species examined when using an unspecific radioimmunoassay which detects several GnRH forms, including lamprey GnRH-I. In the Japanese flounder brain extract, a fourth GnRH immunoreactive fraction was detected with the unspecific radioimmunoassay which did not co-elute with any of the six synthetic GnRH standards used in the present study.     

Self-feeding activity of a pleuronectiform fish, the barfin flounder

The self-feeding activity of the barfin flounder (Verasper moseri) was examined under natural photoperiod and temperature. The experiment, carried out over 12 weeks from September to December, involved tagged fish (mean body weight: 371 g) reared in three 1000-l tanks (3 or 5 fish per tank) with a self-feeder device. The sensor comprised a switch and trigger string with a bead. Self-feeding activity was recorded in all tanks from day 1. The fish fed mostly at night. Strong positive correlations were observed between the number of daily feeder activations and water temperature/photoperiod, and all fish grew during the experiment. These results demonstrate that barfin flounder can operate self-feeding systems and it is suggested that self-feeding activity is influenced by photoperiod and water temperature.     

Traditional seed production in landraces of daikon (Raphanus sativus) in Kyushu, Japan

Landraces of daikon, Japanese radish, Raphanus sativus var. hortensis, are grown and used traditionally in Kyushu, Japan. These landraces are maintained by seeds harvested from selected plants which are transplanted as dormant tap roots or are retained in a portion of field. Seeds kept in intact siliquas on the stems are harvested and stored until use, by hanging the dried stems. Although the relatively small number of plants used in a patch for seed production might be expected to result in a narrow base of genetic diversity of a farmers' lot in a landrace, addition of a few plants with contrasting features, pollen exchange between household patches, and contamination from other cultivars and feral forms may save the landraces from the rapid loss of diversity and deterioration caused by inbreeding depression. The seed production by transplanting described in medieval Japanese literature is relatively similar to the current practice. The factors responsible for maintenance of the landrace together with the cultural and climatic diversity of Japan may have contributed to a high diversity of landraces of the daikon. This revised version was published online in July 2006 with corrections to the Cover Date.     

A natural two-nucleotide deletion affirms role for NADPH oxidase in pathogenicity and perithecium formation in Fusarium graminearum species complex

Members of the Fusarium graminearum species complex are known to cause head blight of wheat. A natural nonpathogenic mutant, strain Fa0233007, was found in the strain collection of the Japanese F. graminearum species complex. In addition, this strain cannot form perithecia. Complete linkage of these deficiencies in the progenies obtained from crossing a wild-type strain and the mutant strain suggested that a single genomic locus was responsible for deficiencies in both traits. The locus was mapped to chromosome 1 using sequence-tagged markers. An NADPH oxidase gene (NoxA, FGSG00739) was identified in the mapped region, and a 2-bp deletion that results in premature termination of the open reading frame was detected in the gene. Both pathogenicity and perithecium formation were recovered by FGSG00739 complementation in the mutant strain.     

Reversible control of hydrogenation of a single molecule

Low-temperature scanning tunneling microscopy was used to selectively break the N-H bond of a methylaminocarbyne (CNHCH3) molecule on a Pt(111) surface at 4.7 kelvin, leaving the C-H bonds intact, to form an adsorbed methylisocyanide molecule (CNCH3). The methylisocyanide product was identified through comparison of its vibrational spectrum with that of directly adsorbed methylisocyanide as measured with inelastic electron tunneling spectroscopy. The CNHCH3 could be regenerated in situ by exposure to hydrogen at room temperature. The combination of tip-induced dehydrogenation with thermodynamically driven hydrogenation allows a completely reversible chemical cycle to be established at the single-molecule level in this system. By tailoring the pulse conditions, irreversible dissociation entailing cleavage of both the C-H and N-H bonds can also be demonstrated.     

Characterization of the neurotoxin produced by isolates associated with avian botulism

Several varieties of birds are affected by type C botulism. We conducted neutralization tests of culture supernatants of isolates from cases of avian botulism. Whereas the toxin produced by isolates derived from mammalian botulism was neutralized only with type C antitoxin, the toxins of all isolates related to avian botulism were neutralized with both type C and D antitoxins. An analysis of nucleotide sequences with several strains revealed that the neurotoxin gene in the isolates from avian botulism comprises two thirds of the type C neurotoxin gene and one third of the type D neurotoxin gene. This indicates that the neurotoxin of avian isolates is a mosaic of type C and D neurotoxins. We prepared three sets of primers to differentiate the gene for the mosaic form from the conserved genes of type C and D neurotoxins. The results of polymerase chain reaction with these primers indicated that all avian botulism-related isolates and specimens possess the gene for the mosaic form of the neurotoxin. The toxins purified from avian and mammalian isolates exhibited the same degree of lethality in mice, but the former showed greater toxicity to chickens than the latter. These results indicate that the mosaic neurotoxin is probably a pathogenic agent causing some forms of avian botulism.