A novel allele of fission yeast rad11 that causes defects in DNA repair and telomere length regulation

Replication protein A (RPA) is a heterotrimeric single-stranded DNA-binding protein involved in DNA replication, recombination and repair. In Saccharomyces cerevisiae, several mutants in the RFA1 gene encoding the large subunit of RPA have been isolated and one of the mutants with a missense allele, rfa1-D228Y, shows a synergistic reduction in telomere length when combined with a yku70 mutation. So far, only one mutant allele of the rad11⁺ gene encoding the large subunit of RPA has been reported in Schizosaccharomyces pombe. To study the role of S.pombe RPA in DNA repair and possibly in telomere maintenance, we constructed a rad11-D223Y mutant, which corresponds to the S.cerevisiae rfa1-D228Y mutant. rad11-D223Y cells were methylmethane sulfonate, hydroxyurea, UV and γ-ray sensitive, suggesting that rad11-D223Y cells have a defect in DNA repair activity. Unlike the S.cerevisiae rfa1-D228Y mutation, the rad11-D223Y mutation itself caused telomere shortening. Moreover, Rad11-Myc bound to telomere in a ChIP assay. These results strongly suggest that RPA is directly involved in telomere maintenance.     

Complete Genome Sequence Analysis of Two Pseudomonas plecoglossicida Phages,Potential Therapeutic Agents

Pseudomonas plecoglossicida is a lethal pathogen of ayu (Plecoglossus altivelis) in Japan and is responsible for substantial economic costs to ayu culture. Previously, we demonstrated the efficacy of phage therapy against P. plecoglossicida infection using two lytic phages (PPpW-3 and PPpW-4) (S. C. Park, I. Shimamura, M. Fukunaga, K. Mori, and T. Nakai, Appl Environ Microbiol 66:1416–1422, 2000, http://dx.doi.org/10.1128/AEM.66.4.1416-1422.2000; S. C. Park and T. Nakai, Dis Aquat Org 53:33–39, 2003, http://dx.doi.org/10.3354/dao053033). In the present study, the complete genome sequences of these therapeutic P. plecoglossicida phages were determined and analyzed for deleterious factors as therapeutic agents. The genome of PPpW-3 (myovirus) consisted of 43,564 bp with a GC content of 61.1% and 66 predicted open reading frames (ORFs). Approximately half of the genes were similar to the genes of the Escherichia coli phage vB_EcoM_ECO1230-10 (myovirus). The genome of PPpW-4 (podovirus) consisted of 41,386 bp with a GC content of 56.8% and 50 predicted ORFs. More than 70% of the genes were similar to the genes of Pseudomonas fluorescens phage ϕIBB-PF7A and Pseudomonas putida phage ϕ15 (podoviruses). The whole-genome analysis revealed that no known virulence genes were present in PPpW-3 and PPpW-4. An integrase gene was found in PPpW-3, but other factors used for lysogeny were not confirmed. The PCR detection of phage genes in phage-resistant variants provided no evidence of lysogenic activity in PPpW-3 and PPpW-4. We conclude that these two lytic phages qualify as therapeutic agents.     

Chromosome mapping of MHC class I in rainbow trout (Oncorhynchus mykiss)

The major histocompatibility complex (MHC) is well-studied in mammals. Much research has addressed the genomic organisation of MHC genes and it is well established that human MHC class I genes are located on chromosome 6. However, information on the organisation of the MHC complex in rainbow trout is only beginning to become available. In the present study it was determined that rainbow trout MHC class I sequences are located on chromosome 18. This is the first reported use of fluorescence in situ hybridisation (FISH) to identify the chromosomal location of genes involved in the immune system of fish.     

Chloroplasts in seeds and dark-grown seedlings of lotus

In most higher plants, mature dry seeds have no chloroplasts but etioplasts. Here we show that in a hydrophyte, lotus (Nelumbo nucifera), young chloroplasts already exist in shoots of mature dry seeds and that they give rise to mature chloroplasts during germination, even in darkness. These shoots contain chlorophyll and chlorophyll-binding proteins CP1 and LHCP. The unique features of chloroplast formation in N. nucifera suggest a unique adaptive strategy for seedling development correlated with the plant's habitat.     

Development and characterization of 24 polymorphic microsatellite markers for bluegill sunfish, <Emphasis Type="Italic">Lepomis macrochirus</Emphasis>

The bluegill sunfish (Lepomis macrochirus) is a notorious exotic fish, which causes serious damages to freshwater ecosystems in the world. Therefore, it is a matter of urgency to control this species for the conservation of ecosystem. New microsatellite loci were developed for L. macrochirus using high throughput 454 GS-FLX+ pyrosequencing. We selected 24 primer pairs that were tested on 47 individuals from Lake Biwa, which encompasses the oldest and largest population of L. macrochirus in Japan. Allele per locus varied 2–17. Expected and observed heterozygosities ranged from 0.185 to 0.888 and 0.120 to 0.872, respectively. Applied to L. macrochirus in Lake Biwa, these markers could detect genetic differentiation among populations, which showed a pattern of isolation by distance. These markers are expected to be useful in the estimation of population structure and ecology of L. macrochirus.     

Mating Type Interaction in Closterium peracerosum-strigosum-littorale: Mating Induced Protoplast Release

Mating type interaction between heterothallic strains of Closteriumperacerosum-strigosum- littorale was studied during the conjugationprocess. When vegetative cells of opposite mating types weremixed under the nitrogen depleted mating conditions, the formationof conjugation-papilla and the release of the protoplast withinthe distended papilla from the gametangial cell were shown inboth the paired and unpaired cells. The protoplasts of pairedcells fused to form zygotes, while those of unpaired cells broke.When cells were mixed at a ratio of around 5 to 3 (mating typeplus to mating type minus), sexual activation occurred at thehighest frequency. Mixing at a ratio of 1 to 3, however, inducedthe highest frequency of disrupted cells. Mating type plus cellswere found to be broken specifically when direct contact withmating type minus cells was prevented. This disruption seemedto be mediated by a factor secreted from mating type minus cells. (Received March 20, 1981; Accepted August 18, 1981)     

Effect of floral organ sizes on female reproductive success in<Emphasis Type="Italic">Erythronium japonicum</Emphasis> (Liliaceae)

We examined the effects of floral organ size on female reproductive success in self-incompatibleErythronium japonicum. We measured tepal size and anther-stigma separation and investigated the relationship between these measurements and fruiting and seeding success. We found that tepal length was positively correlated with fruiting success and the number of seeds per fruit This suggests that pollinator attraction is affected by tepal length in f.japonicum and that the number of pollinator visits affects female reproductive success. Anther-stigma separation was the most variable floral trait measured and was not correlated with either fruiting or seeding success in this species, suggesting that the proportion of outcross pollen deposited on stigmas by pollinators does not increase with anther-stigma separation. This is inconsistent with a previous report onEtyyhtonium. grandiflorum. Pollinator size might explain this interspecific difference in the effect of anther-stigma separation on female reproductive success.     

Influence of isoflurane concentration and hypoxia on functional magnetic resonance imaging for the detection of bicuculline-induced neuronal activation

Our present study shows that transient changes in relative cerebral blood volume (rCBV) induced by stimulation with bicuculline under six different conditions of anesthesia can be detected with high spatial resolution functional magnetic resonance imaging (fMRI). Bicuculline was administered at a low dose to induce neural activation, and no seizure activity was noted. Of the six conditions, the maximal reaction to bicuculline was observed under 1.5% isoflurane in 60% nitrogen and 40% oxygen. Our results imply that the rCBV changes under this level of isoflurane anesthesia with body temperature maintained at 37.5 degrees C are probably suitable for further fMRI studies.     

Differences in distribution patterns around river confluences among hydrophilic vegetation groups

In riparian areas, the distribution patterns of plant species are generally considered to depend on their flooding tolerance. Areas around river confluences are known to experience frequent and/or strong flooding events and provide diverse habitats for plants in riparian areas. However, the degree to which hydrophilic vegetation types increase their distribution around confluences may depend on their flooding tolerance. To test this hypothesis, we compared patch numbers and total areas of ten vegetation groups between confluences and single-flow areas. The vegetation groups were classified on the basis of life form and morphology of dominant species. Additionally, we compared total area of natural bare ground (an index of flooding disturbance) between confluences and single-flow areas. We found that patch numbers of annual grass, forb, and vine, perennial grass and forb, and riparian forest vegetation, as well as total areas of annual forb and vine, perennial grass and forb, bamboo and riparian forest vegetation, and natural bare ground, were greater around river confluences than in single-flow areas. On the other hand, patch numbers of shrub vegetation and total areas of annual grass, perennial vine, willow, and shrub vegetation decreased around confluences. These results suggest that confluences enhance diverse, but not all, types of habitat for hydrophilic vegetation. Thus, river confluences are a key element in maintaining diverse riparian vegetation.