Worldwide migration of parasitic mites as a result of bumblebee commercialization

We investigated the status of infestation by a tracheal mite, Locustacarus buchneri, in natural populations of a Japanese native bumblebee species, Bombus hypocrita, collected on Hokkaido Island and in the Aomori prefecture between 1997 and 2001. We also investigated mite infestation in commercial colonies of the European bumblebee, Bombus terrestris, imported from the Netherlands and Belgium, and the Japanese native species, B. ignitus, imported from the Netherlands, between 1997 and 2001. We detected mites in natural populations of the two B. hypocrita subspecies and in the commercial colonies. Analysis of variations in 535 bp sequences of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene showed that the mite haplotypes in the native populations and in the imported colonies did not overlap in 1997–1999, but in 2000–2001 some mites possessing European CO1 haplotypes were detected in the natural populations of Japanese native bumblebees. In addition, many mites possessing Japanese haplotypes were detected in the imported commercial colonies from Europe. Considering the fact that the Japanese native bumblebees, B. hypocrita, were once exported to Europe for commercialization, these results suggest that bumblebee commercialization has caused overseas migration and cross-infestation of parasitic mites among natural and commercial colonies. However, because the Japanese and European CO1 haplotypes were closely related, there was a possibility that the European haplotypes found in the mites in the Hokkaido Island revealed native variation. To clarify the status of mite invasion, further detailed analysis of genetic variation of the mite, using other genetic markers on additional samples, need to be performed.     

Endozoochorous seed dispersal by sympatric mustelids, Martes melampus and Mustela itatsi, in western Tokyo, central Japan

We investigated seed dispersal by two sympatric mustelid species, the Japanese marten (Martes melampus) and Japanese weasel (Mustela itatsi), along an intercity forest path in western Tokyo, central Japan, from Jul 2007 to Jul 2008. We aimed to investigate the effect of food/habitat preference of these mustelids (martens are semi-arboreal frugivores while weasels are terrestrial carnivores) on their seed dispersal characteristics, which determine their efficacy as seed dispersers. In total, we analyzed 478 fecal samples collected from the two mustelids (N_marten = 381, N_weasel = 97). The proportions of feces containing seeds for martens and weasels were 81.4% and 55.7%, respectively. The number of plant species whose seeds were found within the feces were 28 and 17, respectively. Almost all seeds within feces of both mustelids were intact. The number of plant species whose seeds were found within a single fecal sample ranged from one to four, but no significant difference was detected between the two mustelids. However, marten feces contained a significantly greater number of seeds of most plant species as well as total number of seeds than did weasel feces. The numbers of plant species and seeds represented in marten feces varied seasonally, but those represented in weasel feces did not. Our findings suggest the possibility that both mustelids act in some ways as seed dispersers, although martens seem to disperse a greater diversity and total amount of seeds.     

Antimicrobial susceptibility profile of Acinetobacter species isolated from blood cultures in two Japanese university hospitals

Carbapenem‐resistant Acinetobacter baumannii has rapidly spread worldwide. This study investigated antibiotic susceptibility and genotypic resistance of 123 consecutive blood culture isolates of Acinetobacter species collected between 2003 and 2011 in two Japanese hospitals. The isolates were assigned to 13 species. Carbapenem resistance was detected in four isolates. Only one A. baumannii isolate had bla_OXA‐23 together with ISAba1; the remaining three isolates had IMP‐1 metallo‐β‐lactamase. Quinolone resistance was detected in five isolates that had point mutations in the quinolone resistance‐determining region. The predominance of various non‐A. baumannii species and low prevalence of carbapenem resistance among blood culture isolates of Acinetobacter species in two Japanese hospitals were confirmed.     

Molecular phylogeny and divergence time of the water penny genus Eubrianax (Coleoptera: Psephenidae) in Japan

The phylogenetic relationships among the Japanese members of the genus Eubrianax (Coleoptera: Psephenidae) were examined using the mitochondrial cytochrome oxidase subunit I (COI) gene and nuclear 28S rRNA gene sequences. Based on the molecular phylogeny as well as morphological features, the species status of Eubrianax brunneicornis Nakane, 1952 was proposed. The phylogenetic analyses recovered monophyly of the previously proposed pellucidus species group with four Japanese species, whereas a single Japanese species of the granicollis group was included in the lineage of the ramicornis group with five Japanese species. The divergence times of the species were estimated by dating the phylogenetic tree against the fossil record and a molecular clock based on the COI gene. The divergence of the Japanese species was inferred to have occurred during the Pliocene epoch.     

Influence of ectomycorrhizal fungi on the response of Sitka spruce and Japanese larch to forms of phosphorus

Ectomycorrhizal fungi (Paxillus involutus, Suillus grevillei and two unidentified basidiomycetes from excised Sitka spruce mycorrhizas) were isolated from stands of Sitka spruce either in monoculture or in a mixture with Japanese larch in an Irish conifer plantation. The growth of these fungi and their mycorrhizal formation in Sitka spruce and Japanese larch were examined after incubation in modified Melin-Norkrans medium containing either KH₂PO₄, Ca₃(PO₄)₂ or Fe phytate as the phosphorus (P) source. P. involutus and S. grevillei utilized all three P sources. The unidentified basidiomycetes had limited ability to utilize Fe phytate. Basidiomycete 1 showed poor growth on KH₂PO₄ whereas growth of basidiomycete 2 was low on Ca₃(PO₄)₂. Pure culture synthesis studies confirmed that P. involutus and the two basidiomycetes formed mycorrhizas with both tree species but S. grevillei was mycorrhizal only on Japanese larch. P. involutus formed more mycorrhizas in both conifers than the other fungi. Following inoculation with each of the four fungi, shoot and root dry mass of both Sitka spruce and Japanese larch seedlings was enhanced compared with uninoculated/nonmycorrhizal controls. On Fe phytate, Paxillus-inoculated Sitka spruce seedlings had the lowest primary root length and on KH₂PO₄, Suillus-inoculated Japanese larch had the greatest number of short roots. The only differences when Sitka spruce seedlings were grown in either monoculture or in a mixture with Japanese larch mycorrhizal with S. grevillei were primary root length and number of short roots after growth on media containing Fe phytate.     

Plant species-specific changes in root-inhabiting fungi in a California annual grassland: responses to elevated CO2 and nutrients

Five co-occurring plant species from an annual mediterranean grassland were grown in monoculture for 4 months in pots inside open-top chambers at the Jasper Ridge Biological Preserve (San Mateo County, California). The plants were exposed to elevated atmospheric CO₂ and soil nutrient enrichment in a complete factorial experiment. The response of root-inhabiting non-mycorrhizal and arbuscular mycorrhizal fungi to the altered resource base depended strongly on the plant species. Elevated CO₂ and fertilization altered the ratio of non-mycorrhizal to mycorrhizal fungal colonization for some plant species, but not for others. Percent root infection by non-mycorrhizal fungi increased by over 500% for Linanthus parviflorus in elevated CO₂, but decreased by over 80% for Bromus hordeaceus. By contrast, the mean percent infection by mycorrhizal fungi increased in response to elevated CO₂ for all species, but significantly only for Avena barbata and B. hordeaceus. Percent infection by mycorrhizal fungi increased, decreased, or remained unchanged for different plant hosts in response to fertilization. There was evidence of a strong interaction between the two treatments for some plant species and non-mycorrhizal and mycorrhizal fungi. This study demonstrated plant species- and soil fertility-dependent shifts in below-ground plant resource allocation to different morpho-groups of fungal symbionts. This may have consequences for plant community responses to elevated CO₂ in this California grassland ecosystem. Received: 2 June 1997 / Accepted: 22 August 1997     

Identification of larch species (Larix decidua, Larix kaempferi and Larix X eurolepis) and estimation of hybrid fraction in seed lots by RAPD fingerprints

Species-specific RAPD markers were used to identify the different larch species (Larix decidua and Larix kaempferi) and their interspecific hybrid (Larix X eurolepis). Although morphological differences between pure species and the hybrids exist, differentiation is not always possible, especially at an early stage (seed or plantlet). Eleven RAPD markers differentiated the two larch species, and 4 species-specific markers were sufficient to estimate the F₁ hybrid fraction in a seed lot. The species-specific markers were tested on individual trees of European and Japanese larches of diverse geographic origins and on several seed lots of different origins (F₁, F₂ hybrids and pure species). The 4 specific markers found for the European larch and the Japanese larch were monomorphic and present in all provenances and in all F₁ hybrid trees tested. Polymorphic SCAR fragments were obtained for 3 of the 11 fragments originally selected for the RAPD screening phase. For 2 of them, the sequence had some homology with the mitochondrial genome of other organisms and is thus mitochondrial. The two mitochondrial fragments and the OPF-13₁₀₀₀ fragment exhibited one polymorphic band, thereby maintaining its species-specific identity: OPF-13₁₀₀₀ is specific to the European larch. The 4 RAPD primers selected in this study offer a reliable, quick and cheap tool for the identification of different larch species (Larix decidua and Larix kaempferi) and their interspecific hybrid (Larix X eurolepis). Received: 28 February 1999 / Accepted: 29 April 1999     

Japanese macaque microsatellite PCR primers for paternity testing

Capture and blood sampling in wild primate populations are difficult. For this reason, we need to use DNA extracted from the hair or feces of target animals. The polymerase chain reaction (PCR) method, which amplifies small volumes of DNA, provides an ideal means for studying DNA variations in wild populations. Three sets of PCR primers which amplify highly polymorphic (GT/AC)_n dinucleotide repetitive regions were synthesized from DNA sequences of Japanese macaques (Macaca fuscata). One of the primer pairs detected at least seven alleles in one captive Japanese macaque group. Also, the fathers of four offspring whose mothers had died in a captive group of Japanese macaques were identified. In such cases, the father cannot be determined by the previous DNA fingerprinting method based on the polymorphism of minisatellite DNA. These primers were further tested with some species of the Cercopithecidae, e.g. grivet monkeys (Cercopithecus aethiops tantalus) and hamadryas baboons (Papio hamadryas). The results obtained suggest that these primers can detect stably inherited polymorphic regions in each species.     

New set of microsatellite loci isolated in apricot

We report 99 simple sequence repeats (SSRs) newly isolated from an apricot (Prunus armeniaca L.) genomic library enriched for AG/CT repeats. Twenty SSRs were screened for their polymorphism in 16 apricot cultivars. The number of alleles ranged from two to nine, whereas the expected heterozygosity (H_E) ranged from 0.26 to 0.82. The same SSRs showed also an appreciable transportability across different Prunus species, such as peach, nectarine, almond, European plum, Japanese plum, sweet cherry and sour cherry, with 20% of primers giving successful amplifications in all Prunus species assayed. None gave amplification in apple.     

Characterization of 12 polymorphic microsatellite loci in the Japanese bush warbler Cettia diphone

Japanese bush warblers, Cettia diphone, are a common species in Japan and have a polygynous breeding system. In the breeding ground some males have their own territories with more than one female. Male floaters are also not uncommon in the breeding ground. To understand breeding strategy in this species, exact parentage should be elucidated. In order to obtain a tool for this purpose, we isolated 34 microsatellite loci from a genomic library in this species and developed primers for 12 loci. These primers were tested in the Japanese bush warbler and successfully amplified. In analyses of 49 unrelated individuals, allelic numbers ranged from two to 22, and observed heterozygosity (H_O) ranged from 0.27 to 0.854 except for two loci (Cdi29 and Cdi35a) with H_O < 0.1.     

Optimal stomatal conductance in relation to photosynthesis in climatically contrasting Eucalyptus species under drought

Models of stomatal conductance (g_s) are based on coupling between g_s and CO₂ assimilation (A_net), and it is often assumed that the slope of this relationship (‘g₁’) is constant across species. However, if different plant species have adapted to different access costs of water, then there will be differences in g₁ among species. We hypothesized that g₁ should vary among species adapted to different climates, and tested the theory and its linkage to plant hydraulics using four Eucalyptus species from different climatic origins in a common garden. Optimal stomatal theory predicts that species from sub‐humid zones have a lower marginal water cost of C gain, hence lower g₁ than humid‐zone species. In agreement with the theory that g₁ is related to tissue carbon costs for water supply, we found a relationship between wood density and g₁ across Eucalyptus species of contrasting climatic origins. There were significant reductions in the parameter g₁ during drought in humid but not sub‐humid species, with the latter group maintaining g₁ in drought. There are strong differences in stomatal behaviour among related tree species in agreement with optimal stomatal theory, and these differences are consistent with the economics involved in water uptake and transport for carbon gain.     

Fruiting phenology of animal-dispersed plants in response to winter migration of frugivores in a warm temperate forest on Yakushima Island, Japan

The fruiting phenology of animal-dispersed plants was observed in a warm temperate, evergreen forest on Yakushima Island. The number of ripe fruits was counted for 22 trees, four lianas and one parasitic epiphyte species with sapfruit. These fruits were consumed by birds and Japanese macaques (Macaca fuscata yakui). Birds with small gapes (e.g. Japanese white-eye [Zosterops japonica]) consumed only small fruit less than 6 mm in diameter, while birds with large gapes (e.g. red-capped green pigeon [Sphenurus formosae]) and Japanese macaques consumed a wide range of fruits from 4 to 16 mm in diameter. The larger animals did not ignore the smaller fruits. Brown-eared bulbul (Hypsipetes amaurotis) and Japanese white-eye were the main consumers of sapfruit in terms of frequency in winter. Some of the observed consumers were year-round residents, but most of the consumers migrated to Yakushima Island from the main islands of Japan to overwinter (from November to March), and their abundance in winter was four times as high as during the rest of the year (from May to October). In 23 of the 27 plant species investigated, sapfruit production coincided with their immigration season, whereas tree species bear capsules and nuts during autumn from September to November. We suggest that sapfruit species set their ripe period to the season when frugivorous birds are most abundant.     

The maximal body mass–area relationship in island mammals

Aim A positive power relationship between maximal body mass and land area has previously been reported of the form M_max ∝ Area^0.5 whilst allometric scaling theory predicts either M_max ∝ Area^1.33 or M_max ∝ Area¹. We provide an analysis of the maximal mass–area relationship for four island systems, to test the hypothesis that community relaxation following isolation converges in each case to a slope of Area^0.5. Location Islands of the Japanese archipelago, the western Mediterranean, the Sea of Cortés and Southeast Asia. Methods We calculated the relationship between island area and the maximal body mass of the largest mammal species on the island using linear regression models with log‐transformed variables, and tested the hypothesis that the slopes were not significantly different from 0.5. Results We found a slope of 0.47 within the Japanese archipelago, 0.42 for western Mediterranean islands, 0.73 for the Sea of Cortés islands and 0.50 for Southeast Asian islands. None of these slopes were significantly different from 0.5. Main conclusions Our results provide further empirical support for previous findings of a general maximal body mass–area relationship of M_max ∝ Area^0.5, but they deviate from theoretical predictions. We hypothesize that this mass–area relationship was the ultimate end point of community relaxation initiated by the isolation of the mammal communities. Maximal body mass on each island today probably reflects the interaction between energetic constraints, home range size and island area.     

Molecular phylogeny of the genus<Emphasis Type="Italic">Hypericum</Emphasis> (Hypericaceae) from Korea and Japan: evidence from nuclear rDNA ITS sequence data

As part of our ongoing phylogenetic study of genusHypericum, nuclear ribosomal DNA internal transcribed spacer sequences were analyzed for 36 species ofHypericum as ingroup and two species ofThornea as outgroup. This sampling included most of the previously described species from both Korea and Japan. The ITS phylogeny suggested that the surveyedHypericum species belong to a monophyletic section,Trigynobrathys, and a polyphyletic section,Hypericum. In addition, two monotypic sections,Sampsonia andRoscyna, were identified. Members of sectionHypericum occur in four different lineages worldwide, which imply at least four independent origins. The Korean and Japanese species of sectionHypericum form a monophyletic group, except forH. vulcanicum. Instead, that particular species belongs to a distinct monophyletic group withH. scoreri andH. formosa from other geographic areas, and is a sister to sectionTrigynobrathys. The Korean and Japanese species of sectionTrigynobrathys show a monophyletic origin.H. sampsonii is now recognized as a distinct section rather than being a member of sectionsHypericum orDrosocarpium, as had been indicated previously. Our results differ somewhat from those of recent morphological and cytological studies. The phylogenetic relationships among Korean and Japanese species have now been mostly resolved via ITS phylogeny.     

Molecular genetic mapping of dwarfing genes in oat

 Restriction fragment length polymorphism (RFLP) analysis provides a valuable tool for characterizing and understanding relationships among genes for useful traits in crop species, particularly in ones with complex genomes such as the hexaploid cultivated oat Avena sativa L. (2n=6x=42). Using Bulked Segregant Analysis (BSA) and F₂ RFLP linkage data, we mapped three dominant oat dwarfing loci to different regions of the oat genome. Dw6, in oat line OT207, is 3.3±1.3 cM from the Xumn145B locus, which has not been placed on the hexaploid oat linkage map. Dw7, in line NC2469-3, is 4.3±2.3 cM from Xcdo1437B and 33±4.1 cM from Xcdo708B. This places Dw7 to linkage group 22. Dw8, in the Japanese lines AV17/3/10 and AV18/2/4, mapped 4.9±2.2 cM from Xcdo1319A in an AV17/3/10בKanota’ F₂ population and 6.6±2.6 cM from it in an AV18/2/4בKanota’ population. This places Dw8 to linkage group 3. Aneuploid analysis of markers linked to the dwarfing genes located Dw6 on the smallest oat chromosome (chromosome 18) and Dw7 on the longest satellited chromosome (chromosome 19). The RFLP markers closely linked to the three dwarfing genes identify distinct regions of the oat genome that contribute to plant height and they should be useful in characterizing new genetic sources of dwarfness in oat. Received: 8 May 1997 / Accepted: 20 May 1997     

The major carotenoid in all known species of heliobacteria is the C30 carotenoid 4,4′-diaponeurosporene, not neurosporene

The carotenoids of five species of heliobacteria (Heliobacillus mobilis, Heliophilum fasciatum, Heliobacterium chlorum, Heliobacterium modesticaldum, and Heliobacterium gestii) were examined by spectroscopic methods, and the C₃₀ carotene 4,4′-diaponeurosporene was found to be the dominant pigment; heliobacteria were previously thought to contain the C₄₀ carotenoid neurosporene. In addition, trace amounts of the C₃₀ diapocarotenes diapolycopene, diapo-ζ-carotene, diapophytofluene, and diapophytoene were also found. Up to now, diapocarotenes have been found in only three species of chemoorganotrophic bacteria, but not in phototropic organisms. Furthermore, the esterifying alcohol of bacteriochlorophyll g from all known species of heliobacteria was determined to be farnesol (C₁₅) instead of the usual phytol (C₂₀). Heliobacteria may be unable to produce geranylgeranyol (C₂₀). Received: 10 March 1997 / Accepted: 3 June 1997     

Relationship between parental chromosomic contribution and nuclear DNA content in the coffee interspecific hybrid C. pseudozanguebariae×C. liberica var ‘dewevrei’

 F₁ hybrids were obtained between two coffee species with the same chromosome number (2n=22) but with different nuclear DNA contents [C. pseudozanguebariae (PSE) 2C=1.13 pg and C. liberica var ‘dewevrei’ (DEW) 2C=1.42 pg]. G2 hybrids were obtained by open-pollination of the F₁ hybrids. Genomic in situ hybridisation (GISH) and flow cytometry were used on six F₁ hybrids and seven G2 hybrids to determine their parental chromosomic contribution and their nuclear DNA content (qDNA), respectively. GISH efficiently identified chromosomes from both species. F₁ hybrids had a qDNA intermediate between that of the parental species and contained the expected 11 chromosomes from each species. There was a linear relationship between the number of PSE chromosomes and the nuclear DNA content, which indicates that flow cytometry can be used to give a rough estimate of the parental chromosomic contribution in G2 hybrids. Received: 1 August 1997/Accepted: 25 August 1997     

Geographical variation in female sex pheromones of the rice leaffolder moth, Cnaphalocrocis medinalis: identification of pheromone components in Japan

Sex pheromone components of the Japanese rice leaffolder moth, Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae) were identified from ovipositor extracts of virgin females as (Z)-11-octadecenal, (Z)-13-octadecenal, (Z)-11-octadecen-1-ol and (Z)-13-octadecen-1-ol at a ratio of 11:100:24:36 by GC-EAD, GC, GC-MS. The total amount was estimated to be ca.0.9 ng/female. Field bioassays in Kagoshima, Japan, showed that the two aldehydes are essential for male attraction and the alcohols may have a synergistic effect on the aldehydes. A rubber septum containing 0.9 mg of the four components at the natural ratio was shown to be an effective lure for monitoring this pest in Japan. The above four components are quite different from the sex pheromone components reported previously for the same species of either Philippine or Indian origin; components were shown to be (Z)-11-hexadecenyl acetate and (Z)-13-octadecenyl acetate at a ratio of 98:2 in the Philippine blend and 1:10 in the Indian blend. Furthermore, in the field tests in Japan, neither the Philippine blend nor the Indian blend showed any attractive activity, while the Japanese blend attracted significant numbers of male moths. These results suggest that there are remarkable geographical variations in the sex pheromone composition of this species or there are several distinct species using different sex pheromone blends.     

FLOW CYTOMETRIC APPLICABILITY OF FLUORESCENT VITALITY PROBES ON PHYTOPLANKTON1

The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein‐AM], 5‐chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2′,7′‐dichlorofluorescein diacetate [H₂DCFDA]; and two membrane probes: bis‐(1,3‐dibutylbarbituric acid) trimethine oxonol [DiBAC₄(3)] and SYTOX‐Green) as vitality stains was tested on live and killed cells of 40 phytoplankton strains in exponential and stationary growth phases, belonging to 12 classes and consisting of four cold‐water, 26 temperate, and four warm‐water species. The combined live/dead ratios of all six probes indicated significant differences between the 12 plankton classes (P < 0.01) and between individual species (P < 0.05). No specific differences were observed among strains of one species, among species or strains from different origin, nor between cells in exponential and stationary growth phase except for FDA. FDA showed a significant (P < 0.05) drop of <20% in fluorescence intensity in stationary cells. Of the four esterase probes, the live/dead ratios of FDA and CMFDA were not significantly different from each other, and both performed better than Calcein‐AM and H₂DCFDA (P < 0.001). Of the two membrane probes, DIBAC₄(3) stained rhodophytes and euglenophytes much better than SYTOX‐Green. The 13 algal strains best stainable (high live/dead ratios) among all six probes belonged to nine genera from six classes of phytoplankton. In conclusion, FDA, CMFDA, DIBAC₄(3), and SYTOX‐Green represent a wide choice of vitality probes in the study of phytoplankton ecology, applicable in many species from different algal classes, originating from different regions and at different stages of growth.     

Genetic diversity in the threatened insular endemic plantAster asa-grayi (Asteraceae)

Genetic diversity in an insular endemic plantAster asa-grayi was examined using enzyme electrophoresis. Distribution ofA. asa-grayi is restricted to only four subtropical islands of Japan, and this species is listed as vulnerable to extinction in the Red Data Book of Japanese wild plants. A total of 161 individuals were sampled from five populations on four islands. Genetic diversity values at the population level were very low, compared to other plant species with a similar life history. Genetic variability at the species level is comparable to the mean value of endemic species. Genetic differentiation among populations is extremely high (G_ST= 0.71), indicating that the gene flow among populations is highly impeded, and pollen and seed dispersal is limited due to the pollinators and the seed morphology. This is because the four islands are geographically isolated. Fixation indices suggested that most populations do not randomly cross. To conserve the genetic diversity of the species, artificial crossings among different island populations are necessary.