<Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>. The most important bacterial warfare agents — <Emphasis Type="Italic">review</Emphasis>

There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes: Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed.     

Host specificity of <Emphasis Type="Italic">Euops chinesis</Emphasis>, a potential biological control agent of <Emphasis Type="Italic">Fallopia japonica</Emphasis>, an invasive plant in Europe and North America

Fallopia japonica (Houttuyn) Ronse Decraene (Polygonaceae) is a serious invasive weed in North American and Europe. In its native China, a leaf-rolling weevil, Euops chinesis (Coleoptera: Attelabidae) was found attacking F. japonica in the field. No-choice tests, multiple-choice tests, open field tests and field surveys were conducted as a measure of its host specificity. Forty-six plant species were selected from 17 families for host range testing, among which, six species, F. multiflora, F. japonica, Persicaria perfoliata, Rumex acetosa, R. japonicus and R. aquaticus, were exposed to adults in no-choice tests. However, larvae could only develop successfully on F. japonica, and this plant appeared to be the only host in the field, suggesting the weevil is host-specific. As larval development appears to depend on a fungus in the leaf rolls, the insect–fungus mutualism and risks including host specificity of the fungus should be evaluated before the insect’s introduction.     

Interaction of <Emphasis Type="Italic">Bdellovibrio bacteriovorus</Emphasis> with bacteria <Emphasis Type="Italic">Campylobacter jejuni</Emphasis> and <Emphasis Type="Italic">Helicobacter pylori</Emphasis>

Interaction of Bdellovibrio bacteriovorus 100NCJB with bacteria Campylobacter jejuni (strains 1, 2, 3, 4, and 5) and Helicobacter pylori, strain TX30a, was confirmed. The results indicate that lytic activity of bdellovibrios both in liquid media and cells attached to a surface was observed. The potential use of the antimicrobial activity of predatory bacteria for environmental bioprotection and public health is discussed.     

Utilization of dibenzothiophene as sulfur source by <Emphasis Type="Italic">Microbacterium</Emphasis> sp. <Emphasis Type="Italic">NISOC-06</Emphasis>

Oil-polluted soils were sampled from National Iranian South Oil Company (NISOC) for isolation and screening of C–S and not C–C targeted Dibenzothiophene (DBT) degrading microorganisms. Microbacterium sp. NISOC-06, a C–S targeted DBT degrading bacterium, was selected and its desulfurization ability was studied in aqueous phase and water-gasoline biphasic systems. The 16srRNA gene was amplified using universal eubacteria-specific primers, PCR product was sequenced and the sequence of nearly 1,500 bp 16srDNA was studied. Based on Gas Chromatography results Microbacterium sp. NISOC-06 utilized 94.8% of 1 mM DBT during the 2 weeks of incubation. UV Spectrophotometry and biomass production measurements showed that the Microbacterium sp. NISOC-06 was not able to utilize DBT as a carbon source. There was no accumulation of phenolic compounds as Gibb’s assay showed. Biomass production in a biphasic system for which DBT-enriched gasoline was used as the sulfur source indicated the capability of Microbacterium sp. NISOC-06 to desulfurize gasoline.     

Mapping species differences for adventitious rooting in a <Emphasis Type="Italic">Corymbia torelliana</Emphasis> × <Emphasis Type="Italic">Corymbia citriodora</Emphasis> subspecies <Emphasis Type="Italic">variegata</Emphasis> hybrid

Quantitative trait loci (QTL) detection was carried out for adventitious rooting and associated propagation traits in a second-generation outbred Corymbia torelliana × Corymbia citriodora subspecies variegata hybrid family (n = 186). The parental species of this cross are divergent in their capacity to develop roots adventitiously on stem cuttings and their propensity to form lignotubers. For the ten traits studied, there was one or two QTL detected, with some QTL explaining large amounts of phenotypic variation (e.g. 66% for one QTL for percentage rooting), suggesting that major effects influence rooting in this cross. Collocation of QTL for many strongly genetically correlated rooting traits to a single region on linkage group 12 suggested pleiotropy. A three locus model was most parsimonious for linkage group 12, however, as differences in QTL position and lower genetic correlations suggested separate loci for each of the traits of shoot production and root initiation. Species differences were thought to be the major source of phenotypic variation for some rooting rate and root quality traits because of the major QTL effects and up to 59-fold larger homospecific deviations (attributed to species differences) relative to heterospecific deviations (attributed to standing variation within species) evident at some QTL for these traits. A large homospecific/heterospecific ratio at major QTL suggested that the gene action evident in one cross may be indicative of gene action more broadly in hybrids between these species for some traits.     

Strict requirement of fluctuating temperatures as a reliable gap signal in <Emphasis Type="Italic">Picris hieracioides</Emphasis> var. <Emphasis Type="Italic">japonica</Emphasis> seed germination

Picris hieracioides var. japonica (Asteraceae), which grows in occasionally disturbed habitats such as riverbanks, is rarely observed under dense vegetation. We examined the effect of the experience and timing of receiving leaf-transmitted light in gap-detecting seed germination in this plant. Seeds under unfiltered light, which simulated the light conditions of seeds on the soil surface in a canopy gap, germinated at a constant temperature of 20°C. However, most seeds in darkness, which simulated the light conditions of seeds buried in the soil without receiving leaf-transmitted light, germinated under temperature fluctuations of over 4°C. Seeds in darkness after receiving leaf-transmitted light for 1 week, which simulated the light conditions of seeds buried in the soil after receiving leaf-transmitted light, germinated under temperature fluctuations of over 8°C. Finally, seeds under continuous leaf-transmitted light, which simulated the light conditions of seeds on the soil surface below preexisting vegetation, germinated under temperature fluctuations of over 12°C. Seeds that experience unfiltered light, which suggests that they are in a gap, should not delay germination. In contrast, seeds that have received leaf-transmitted light should delay germination until the vegetation above is removed. Seeds exposed to leaf-transmitted light required larger temperature fluctuations in darkness than did untreated seeds, and seeds under continuous leaf-transmitted light required the largest temperature fluctuations. The various germination reactions to each gap signal in P. hieracioides var. japonica seeds allow the more reliable detection of gaps for subsequent seedling establishment. The requirement for gap signals that created high precision of timing in the germination process results in the germination of this species only in gaps. Therefore, P. hieracioides var. japonica is rarely found under dense vegetation.     

Frequencies of the <Emphasis Type="Italic">DRB1</Emphasis>, <Emphasis Type="Italic">DQA1</Emphasis>, <Emphasis Type="Italic">DQB1</Emphasis> and <Emphasis Type="Italic">TNFA</Emphasis> alleles in immigrant population of west Siberia

Based on population analysis of the DRB1, DQA1, DQB1 and TNFA allele frequency distribution patterns, regional features of immunogenetic structure of the population of West Siberia were investigated. Statistically significant linkage disequilibrium within the HLA class II region, as well as between the TNFA and DRB1, DQA1, and DQB1 was demonstrated. Population frequency distribution patterns of two- and multilocus haplotypes were examined.     

The seasonal occurrence of endophytic fungus, <Emphasis Type="Italic">Mycosphaerella buna</Emphasis>, in Japanese beech, <Emphasis Type="Italic">Fagus crenata</Emphasis>

The seasonal occurrence of Mycosphaerella buna in leaves and contiguous organs of Fagus crenata was studied in a Japanese beech forest, Ibaraki, Japan, in 1998 and 1999. Mycosphaerella buna was not isolated from newly developed leaves in May, but it was isolated from asymptomatic leaves after June. The frequency of its occurrence gradually increased until leaffall. In contrast, M. buna was not isolated from overwintered buds, leaf petioles, or contiguous current-year twigs. The spermogonia and pseudothecia were observed in dead leaves after leaffall. The mature pseudothecia were found on dead leaves from May to July. The ascospores produced in the pseudothecia were suggested to infect newly developed leaves.Contribution no. 173, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

<Emphasis Type="Italic">In vitro</Emphasis> biosynthesis of strictosidine using<Emphasis Type="Italic">lonicera japonica</Emphasis> leaf extracts and recombinant yeast

Strictosidine is a key intermediate in the biosynthesis of the terpenoid indole alkaloid (T1A) pathway. It results from a condensation reaction, catalyzed by strictosidine synthase (STR), between tryptamine and secologanin. We have now developed a useful method, based on enzyme-assisted synthesis, to produce strictosidine. Our procedure utilizes leaf extracts from Japanese honeysuckleLonicera japonica Thunb. as a secologanin source. In these experiments, an enzyme extract was prepared from transgenic yeastSaccharomyces cerevisiae that expresses theCatharanthus roseus STR (CrSTR) coding region. Strictosidine was then isolated with a 38% yield based on the initial amount of tryptamine in the enzymatic reaction.     

Overexpression of the Chitosanase Gene in <Emphasis Type="Italic">Fusarium solani</Emphasis> via <Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-Mediated Transformation

To overexpress the chitosanase gene (csn) in F. solani, a vector based on pCAMBIA 1300 was constructed. The csn gene, which is under control of the Aspergillus nidulans gpdA promoter and A. nidulans trpC terminator, was introduced back into the F. solani genome by Agrobacterium tumefaciens-mediated transformation, and the herbicide-resistance gene bar from Streptomyces hygroscopicus was used as the selection marker. Transformants which showed a significant increase in chitosanase production (~2.1-fold than control) were obtained. Southern blot analysis indicated that most transformants had a single-copy T-DNA integration.     

Multiple host-ant use by the predatory social parasite <Emphasis Type="Italic">Phengaris</Emphasis> (=<Emphasis Type="Italic">Maculinea</Emphasis>) <Emphasis Type="Italic">arion</Emphasis> (Lepidoptera,Lycaenidae)

Phengaris (=Maculinea) arion is an endangered social parasite of Myrmica ants, and for a very long time was considered as specific to Myrmica sabuleti. Previous studies carried out in Poland suggested some discrepancies within this assumption, and therefore a much more intensive survey was undertaken. The host ant use of P. arion was studied at five sites in different types of biotopes in Poland, i.e. xerothermal grasslands where Thymus pulegioides was used as a larval food plant by the butterfly, and more or less sandy biotopes with Thymus serpyllum. Altogether nine Myrmica species were recorded, and considerable variation in species composition and density of nests was recorded. At four localities M. sabuleti proved to be the most common ant. A total of 529 Myrmica nests were examined, and only 20 of them contained larvae and pupae of P. arion. Host ants belonged to five different species, i.e. M. sabuleti, Myrmica scabrinodis, Myrmica schencki, Myrmica lobicornis and Myrmica hellenica. Only at one site (NE Poland) was a significant heterogeneity in parasitation rates among Myrmica species detected. M. lobicornis was the most often infested ant there, which may suggest local specialisation of the butterfly. Overall low parasitism rates may explain the vulnerability of P. arion in Central Europe but further studies are also necessary.     

<Emphasis Type="Italic">Barleria compacta</Emphasis>: a new species in <Emphasis Type="Italic">Barleria</Emphasis> sect. <Emphasis Type="Italic">Prionitis</Emphasis> (Acanthaceae) from Somalia

Following a re-examination of the material treated under Barleria brevispina (Fiori) Hedrén in the recent Flora of Somalia account of the Acanthaceae, it is concluded that two distinct species are involved and Barleria compacta Malombe & I. Darbysh. is described here from north-eastern Somalia. Its affinities and conservation status are discussed.     

A silvering index for the Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To establish a simple and reliable index for determining silvering stages of the Japanese eel, Anguilla japonica, we observed the colorations of various body parts and biological characteristics of the eels collected in a coastal area of Japan (Mikawa Bay). The four silvering stages are characterized by the colorations of pectoral fins and ventral skin as follows: (1) Y1, yellow eel without a metallic hue at the base of pectoral fins, (2) Y2, late yellow eel with a metallic hue at the base of the pectoral fins but without melanization at the tip of pectoral fins, (3) S1, silver eel with complete melanization at the tip of pectoral fins but without full pigmented belly in black or dark brown, and (4) S2, late silver eel with black or dark brown belly. The body size, eye diameter and sexual maturity of each stage increased in the order of Y1, Y2, S1 and S2 stages, whereas the digestive tract degenerated in the same order, suggesting a sequential development of these ontogenetic stages identified in the study. The Y1, Y2 and S1 stages could be also distinguished by canonical discriminant function analysis using three internal (gonad-somatic index, GSI; hepato-somatic index, HIS; and gut index) and two morphometric (condition factor and eye index) parameters, supporting the significance of these stages. This method of staging for the silvering process of the Japanese eel appeared to be applicable to all specimens of this species, since this index used only simple external characteristics that would be easy to observe during field surveys.