Foliar nutrient concentrations of small Chamaecyparis in Taiwan

Summary Seedlings ofChamaecyparis lawsoniana, C. formosensis andC. taiwanensis were grown in a forest nursery in Taiwan. Native saplings of the latter two species were sampled in a clearcut and in the forest understory. Foliar nutrient concentrations were mostly in the mid-range of values for other conifers. However, Ca (.59–1.5%) and Cu (7–20 ppm) were almost always high. Concentrations of N, P, and K in field saplings in Taiwan were low enough (.7–1.5%, .07–.15%, .47–.60%, respectively) to indicate that these elements are probably limiting growth.Among the three species,C. lawsoniana usually had the lowest foliar nutrient values. When a difference among the species from Taiwan occurred,C. taiwanensis usually was lower. Foliar nutrient concentration varied among provenances, especially withinC. formosensis. Seasonal variability did not occur consistently; nutrients which did change increased their concentrations from the dormant to the growing season, except for K which decreased.     

Population Structure and Historical Demography of the Oriental River Prawn (Macrobrachium nipponense) in Taiwan

The oriental river prawn (Macrobrachium nipponense) is a non-obligatory amphidromous prawn, and it has a wide distribution covering almost the entire Taiwan. Mitochondrial DNA fragment sequences of the cytochrome oxidase subunit I (COI) and 16S rRNA were combined and used to elucidate the population structure and historical demography of oriental river prawn in Taiwan. A total of 202 individuals from six reservoirs and three estuaries were separately collected. Nucleotide diversity (π) of all populations was 0.01217, with values ranging from 0.00188 (Shihmen Reservoir, SMR, northern Taiwan) to 0.01425 (Mingte Reservoir, MTR, west-central Taiwan). All 76 haplotypes were divided into 2 lineages: lineage A included individuals from all sampling areas except SMR, and lineage B included specimens from all sampling locations except Chengching Lake Reservoir (CLR) and Liyu Lake Reservoir (LLR). All F _ST values among nine populations were significantly different except the one between Jhonggang River Estuary (JGE, west-central Taiwan) and Kaoping River Estuary (KPE, southern Taiwan). UPGMA tree of nine populations showed two main groups: the first group included the SMR and Tamsui River Estuary (TSE) (both located northern Taiwan), and the second one included the other seven populations (west-central, southern and eastern Taiwan). Demographic analyses implied a population expansion occurred during the recent history of the species. The dispersal route of this species might be from China to west-central and west-southern Taiwan, and then the part individuals belonging to lineage A and B dispersed southerly and northerly, respectively. And then part individuals in west-central Taiwan fell back to and stay at estuaries as the sea level rose about 18,000 years ago.     

Development of a dot immunobinding assay to detect Phytophthora spp. in naturally dark-rooted woody plants

An indirect dot immunobinding assay (DIBA) with a PAb raised against an isolate of P. cinnumomi was evaluated to detect Phytophthora spp. in naturally dark-rooted woody plants. Screening of different modifications of the procedure were done with root samples of Chamaecyparis lawsoniana non-infected and infected with P. cinnamomi. With the optimised DIBA procedure, root infection could be diagnosed by a clearly defined halo around the spot and by a colour change in the spot using Fast Red or Fast Blue substrate. Detection of different Phytophthora species in Chamaecyparis plants from commercial nurseries was successful with the optimised procedure.     

The commercial exploitation of the white rot fungus Lentinula edodes (shiitake)

The Japanese wood mushroom, Lentinula edodes, can be cultivated under UK climatic conditions using extensive cultivation systems adapted from traditional growing methods developed in Japan and Taiwan. As well as producing a desirable and high value food product, cultivation offers a viable alternative enterprise for farmers. The growing methods described could provide incentives for the management of broad-leaved woodland in Britain with consequent conservation benefits. Four different strains of Lentinula edodes were grown on oak logs under natural environmental conditions below the woodland canopy and fruited in polyethylene tunnels. Yields ranged from 0.5–0.9 kg wet weight shiitake per cordwood log per year over a three year fruiting period.     

Investigation of a gamasid mite infestation in a UK textile mill caused by Dermanyssus gallinae (DeGeer, 1778) (Mesostigmata: Dermanyssidae) special lineage L1

A recurrent mite infestation affecting a room used to inspect fabric in a UK textile mill was investigated to allay concerns of any potential health risks to factory staff, and to inform the unknown risk of downgrading of the product. The approach integrated conventional morphological examination of adult female mites by referring to published identification keys, with molecular speciation based on amplification of a 16S ribosomal DNA fragment. The methods enabled the mites to be unambiguously identified as Dermanyssus gallinae ‘special lineage L1’. Subsequent investigations showed the source of infestation to be pigeons nesting in the air ducts, with the gamasid mites moving into the room once the young birds had fledged. This is the first report of D. gallinae ‘special lineage L1’ in northern Europe. Previous reports of nosocominal gamasoidosis caused by D. gallinae ‘special lineage L1’ originating from feral pigeon populations have been from southern Europe. Confirmation of the mite identity was important in allowing the mill to take remedial and preventive action. In this clinical communication, we provide images of the key morphological features used to identify D. gallinae and describe a molecular protocol to confirm ‘special lineage L1’.     

Effects of environment,seedling age,and seed source on leaf resistance of three species of Chamaecyparis and Tsuga chinensis

Summary The objectives of this study were to determine how stomata of Chamaecyparis spp. react to environmental changes and to determine in what ways leaf resistance patterns may have adaptive value for these confers. Leaf resistances were measured of Chamaecyparis taiwanensis, C. formosensis and C. lawsoniana seedlings in a forest nursery, and of saplings of the first two species and Tsuga chinensis growing in natural forest and clearcuts. Light, temperature, humidity, and sometimes xylem pressure potential were measured concurrently.In the nursery, C. lawsoniana, the species from the driest climate, had the lowest resistance, daily means for well-watered yearold plants being 2–5 s cm^-1. Leaf resistances of well-watered C. formosensis and C. taiwanensis from more humid habitats were two and three times greater. Leaf resistance was more sensitive to environment in those nursery populations of C. formosensis from habitats where a dry season is more severe. There were no clear patterns of variation within the other species. Resistance of scale leaves increased with seedling size and age. Primary leaves of C. formosensis had higher average resistance than its scale leaves, and were more responsive to changes in the environment.In the field, C. taiwanensis had higher resistance than C. formosensis. Leaf resistance of Tsuga chinesis was quite similar to that of C. formosensis. Resistance decreased from winter to spring except during a severe dry spell in March.Changes in leaf resistance were most consistently correlated with humidity, which often increased during the day as humid air rose from the lowlands. Stomata opened as humidity rose even when plant xylem pressure potentials were -15 to -23 bars. Stomatal reaction of Chamaecyparis spp. varies in ways apparently adaptive to their environment.     

Historical biogeography and phylogenetic relationships of the genus Chamaecyparis (Cupressaceae) inferred from chloroplast DNA polymorphism

The genus Chamaecyparis comprises five species and one variety native to Taiwan, Japan, Canada, and USA, which demonstrates a classical eastern Asian, western North American, and eastern North American disjunct distributional pattern. The phylogenetic relationships of the species of Chamaecyparis were inferred by comparing 1130 bp of the combined data set of chloroplast trnV intron and petG-trnP intergenic spacer. The phylogenetic tree shows that Chamaecyparis nootkatensis (Cupressus nootkatensis or Xanthocyparis nootkatensis) is clearly diverged from other Chamaecyparis species. For Chamaecyparis species, C. thyoides is sister to C. formosensis and C. pisifera and these together form a monophyletic group. C. lawsoniana is sister to C. obtusa and C. taiwanensis; and these form another monophyletic group. Homogeneity in evolutionary rates was found among species in these two monophyletic groups. Results indicate the divergent evolution of C. taiwanensis and C. formosensis and molecular evidence in this investigation supports C. taiwanensis as a variety of C. obtusa. Utility of cpDNA intergenic spacer petG-trnP in Chamaecyparis is also discussed. Several biogeographical implications were inferred: (1) at least two divergence events have produced the eastern Asian, and both western and eastern North American disjunct distribution in Chamaecyparis; (2) intercontinental sister species pairs are found in Chamaecyparis; (3) cpDNA divergence between two intercontinental sister pairs of C. thyoides and C. pisifera, and C. lawsoniana and C. obtusa is 2.8% and 1.1%, which suggest an estimated divergence time of 14 and 5.5 million years ago during middle and late Miocene, respectively; (4) cpDNA divergence of two Asian Chamaecyparis groups between C. obtusa and C. taiwanensis, and between C. pisifera and C. formosensis is 0.25% and 0.57%, which suggest an estimated divergence time of 1.3 and 2.9 million years ago during Pleistocene and late Pliocene, respectively; these estimated divergence times suggest a relatively recent migration of Chamaecyparis to Taiwan from the Japanese Archipelago; (5) that climatic deterioration caused the disappearance of Chamaecyparis in continental Asia is probable.     

Isolation and Identification of Pythiaceous Fungi from Irrigation Water and their Pathogenicity to Antirrhinum, Tomato and Chamaecyparis lawsoniana

Pythiaceous fungi were isolated from irrigation water using a variety of natural and artificial baits. Isolates were also obtained by plating water samples directly on the surface of selective agar media. The selective medium of Ocana and Tsaq (1966) PlOVP, was modified by substituting rifampicin and ampicillin (10 and 500 μg cm^?3 respectively) for vancomycin to suppress bacterial growth from water samples. The pythiaceous fungi were identified as Pythiitm dissotocitm, P. middletonii, P. mamillatum, P. rostratum, Pythium“group 1”, “group 2” and “group 3” and Phytophtbom gona-podyides. All isolates of P. gonapodyides were the A1 strain and produced oospores when paired with an A2 isolate of P. drechsleri. Isolates were tested for their pathogenicity to Antirrhinum, tomato and Chatmaecyparis lawsoniana cv. Ellwoodii. Pythium middletonii and Pythium“group 1” caused severe pre-emergence damping-off of Antirrhinum seedlings, P. mamillatuni, P. rostratum and Pythium“group 3” were less pathogenic to the same host while P. dissotocum, Pythium“group 2” and Phytophthora gonapodyides were non-pathogenic. Only isolates of Pythium“group 1” were pathogenic to tomato seedlings. None of the fungi was pathogenic to rooted cuttings of Chamaecyparis lawsoniana cv. Ellwoodii.     

Different dispersal histories of lineages of the earthworm <Emphasis Type="Italic">Aporrectodea caliginosa</Emphasis> (Lumbricidae,Annelida) in the Palearctic

Earthworms are among the most abundant and ecologically important invasive species, and are therefore a good object for studying genetic processes in invasive populations. Aporrectodea caliginosa is one of the most widespread invasive earthworms in the temperate zone. It is believed to have dispersed from Europe to all continents except Antarctica. It is known that A. caliginosa consists of three genetic lineages, and genetic diversity is high both among and between them. We attempted to use that high genetic diversity to study A. caliginosa dispersal in the Palearctic based on a sample of 40 localities ranging from eastern Europe to the Russian Far East, and to compare our data to other studies on this species in western Europe and North America. Two genetic lineages were found in the studied sample. Only negligible decrease in genetic diversity was observed for the lineage 2 of A. caliginosa from West Europe to the Far East, suggesting multiple human-mediated introductions. In contrast, lineage 3 is abundant in West Europe and Belarus, but is absent from the East European Plain, the Urals, and the Far East. However, it is present in West Siberia, where it has greatly reduced genetic diversity, indicating long-distance dispersal accompanied by a bottleneck event. Thus, although these two lineages of A. caliginosa are morphologically indistinguishable, they have dramatic differences in their distributions and dispersal histories.     

Some effects of differently-acting nematicides on the cereal cyst-nematode (Heterodera avenae) and on the appearance of ''scorch'' in spring wheat on light loamy sand

In fungitoxicity tests against Phytophthora cinnamomi on Chamaecyparis lawsoniana cv. Ellwoodii, a drench of furalaxyl (1000 mg a.i./l) applied to the compost in which 1-yr-old plants were growing, 1 wk before they were inoculated with 650 000 zoospores, controlled disease for at least 12 months. With an inoculum dose of 650 zoospores/plant, furalaxyl at 500 mg a.i./l controlled disease even when inoculation was 12 wk after fungicide treatment. Aluminium tris (ethyl phosphonate) (2000 mg a.i./l) applied as a drench 1 wk before inoculation with 650 000 zoospores/plant did not prevent root infection but delayed foliar symptoms for 9 months: the same treatment, using etridiazole (500 mg a.i./l) only slightly reduced disease incidence. When applied as a single drench 2 days before inoculation, prothiocarb (2000 mg a.i./l) and cuprammonium compounds (200 mg a.i./l) were much less effective than furalaxyl (1200 mg a.i./l), sodium ethyl phosphonate (1500 mg a.i./l), aluminium tris (ethyl phosphonate) (1500 mg a.i./l) or etridiazole (500 mg a.i./l). However, a drench of furalaxyl at 1000 mg a.i./l, aluminium tris (ethyl phosphonate) at 2000 mg a.i./l or etridiazole at 500 mg'a.i./l did not eradicate P. cinnamomi from compost containing infected root debris. Pre-planting drenching of the compost was ineffective. All fungicide treatments were non-phototoxic to 1-yr-old C. lawsoniana cv. Ellwoodii. These results are of special relevance to the control of P. cinnamomi on container-grown woody ornamentals.     

Genomic patterns and the evolutionary origin of an invasive fungal pathogen (Hymenoscyphus fraxineus) in Europe

In the 1990s, a highly pathogenic and invasive lineage of the Asian ascomycete Hymenoscyphus fraxineus appeared in Europe. Spreading across the continent, the fungal lineage devastates common ash (Fraxinus excelsior) populations and is a threat to ash species worldwide.In contrast to highly diverse reference populations in Japan and East Russia, the expanding European population is characterized by the persistence of a mosaic of two ancestral haplotypes, and presumed constraints to recombination. Four hypotheses were proposed for the evolutionary origin of the European lineage. The first hypothesis H1 states that two, and only two, pathogenic founder individuals were transferred from Asia to Europe and successfully established. Alternatively, in addition to the fungus arriving in Europe, a change in function might have occurred and only then the lineage became invasive. The cause of such a change might have been H2: one to two mutations during meiosis; H3: the lateral transfer of a European cytoplasmatic element modifying the behavior of H. fraxineus; or H4: a hybridization event with a closely related European ascomycete.Only the hybridization hypothesis seems to explain all of the genome-wide characteristics observed for the nuclear genome. Existing reconstructions of phylogenetic relationships at different taxonomic levels provide the evolutionary context of the lineages potentially involved. Vital information was gained from three configurations of nucleotide diversity and population differentiation associated with the genomic mosaic of monomorphic tracts and diversity islands.This synthesis points to the importance of identifying the taxonomic and geographic origins of the conserved two genomic ancestries within the European lineage of H. fraxineus and of reconstructing the mosaic of ancestral parental haplotypes (ancestry blocks) across the genome. Only with this information at hand will it be possible, to reliably interpret population-genomic data in the search for the functional elements underlying this unbalanced host-pathogen system.     

Evolution of the Euphrasia transmorrisonensis complex (Orobanchaceae) in alpine areas of Taiwan

Aims To unravel isolation and differentiation of the genetic structure of the Euphrasia transmorrisonensis complex, a showy herb, among alpine regions of mountain peaks in subtropical Taiwan and to infer its evolutionary history. Location Alpine ecosystems of high‐montane regions of Taiwan. Methods Phylogenetic analyses of the trnL intron and the trnL–trnF intergenic spacer of chloroplast (cp) DNA, and the intertranscribed spacer (ITS) of nuclear ribosomal (nr) DNA between 18S and 26S were carried out on 18 populations of the E. transmorrisonensis complex in Taiwan. Results In total, 10 haplotypes for cpDNA and 14 haplotypes for nrDNA were detected. Three population groups located in the northern, north‐eastern, and south‐central regions of the Central Mountain Range (CMR) were revealed according to the frequencies of haplotypes and haplotype lineages of nrDNA. Balancing selection might have played a role in the evolution of Euphrasia in Taiwan. Main conclusions By integrating the spatial‐genetic patterns of cpDNA and nrDNA, two possible evolutionary histories of Euphrasia in Taiwan were inferred. The favourable hypotheses for interpreting the data suggest at least three origins of the E. transmorrisonensis complex in Taiwan, corresponding to each nuclear lineage in the northern (II), northern/north‐eastern (I), and central/southern regions (III) with subsequent hybridization between lineages I and II and lineages II and III. These lineage boundaries are strengthened by the finding that haplotypes of C derived from cpDNA were found in the geographical region of lineage II of nrDNA, while haplotypes of A derived from cpDNA were found in the region of lineage III of nrDNA. Thus, the origin of chloroplasts exclusive to lineages II and III supports their long‐term isolation from one another.     

Nested Allele-Specific PCR Primers Distinguish Genetic Groups of Uncinula necator

Isolates of the obligately biotrophic fungus Uncinula necator cluster in three distinct genetic groups (groups I, II, and III). We designed PCR primers specific for these groups in order to monitor field populations of U. necator. We used the nucleotide sequences of the gene that encodes eburicol 14α-demethylase (CYP51) and of the ribosomal DNA internal transcribed spacer 1 (ITS1), ITS2, and 5.8S regions. We identified four point mutations (three in CYP51 and one in ITS1) that distinguished groups I and II from group III based on a sample of 132 single-spore isolates originating from Europe, Tunisia, Israel, India, and Australia. We developed a nested allele-specific PCR assay in which the CYP51 point mutations were used to detect and distinguish groups I and II from group III in crude mildewed samples from vineyards. In a preliminary study performed with samples from French vineyards in which isolates belonging to genetic groups I and III were present, we found that a shift from a population composed primarily of group I isolates to a population composed primarily of group III isolates occurred during the grapevine growing season.     

A relict lineage and new species of green palm-pitviper (Squamata,Viperidae, Bothriechis) from the Chortís Highlands of Mesoamerica

A new species of palm-pitviper of the genus Bothriechis is described from Refugio de Vida Silvestre Texíguat in northern Honduras. The new species differs from congeners by having 19 dorsal scale rows at midbody, a bright green dorsal coloration in adults, the prelacunal scale fused to the second supralabial, and in representing a northern lineage that is sister to Bothriechis lateralis, which is distributed in Costa Rica and western Panama and is isolated from the new taxon by the Nicaraguan Depression. This represents the 15th endemic species occurring in Refugio de Vida Silvestre Texíguat, one of the richest herpetofaunal sites in Honduras, itself being the country with the highest degree of herpetofaunal endemism in Central America. We name this new species in honor of a Honduran conservationist slain in fighting against illegal logging, highlighting the sacrifices of rural activists in battling these issues and the critical importance of conservation in these areas.     

Whole Genome-Sequencing and Phylogenetic Analysis of a Historical Collection of Bacillus anthracis Strains from Danish Cattle

Bacillus anthracis, the causative agent of anthrax, is known as one of the most genetically monomorphic species. Canonical single-nucleotide polymorphism (SNP) typing and whole-genome sequencing were used to investigate the molecular diversity of eleven B. anthracis strains isolated from cattle in Denmark between 1935 and 1988. Danish strains were assigned into five canSNP groups or lineages, i.e. A.Br.001/002 (n = 4), A.Br.Ames (n = 2), A.Br.008/011 (n = 2), A.Br.005/006 (n = 2) and A.Br.Aust94 (n = 1). The match with the A.Br.Ames lineage is of particular interest as the occurrence of such lineage in Europe is demonstrated for the first time, filling an historical gap within the phylogeography of the lineage. Comparative genome analyses of these strains with 41 isolates from other parts of the world revealed that the two Danish A.Br.008/011 strains were related to the heroin-associated strains responsible for outbreaks of injection anthrax in drug users in Europe. Eight novel diagnostic SNPs that specifically discriminate the different sub-groups of Danish strains were identified and developed into PCR-based genotyping assays.     

Genetic Diversity of Clinical and Environmental Vibrio parahaemolyticus Strains from the Pacific Northwest

Since 1997, cases of Vibrio parahaemolyticus-related gastroenteritis from the consumption of raw oysters harvested in Washington State have been higher than historical levels. These cases have shown little or no correlation with concentrations of potentially pathogenic V. parahaemolyticus (positive for the thermostable direct hemolysin gene, tdh) in oysters, although significant concentrations of tdh⁺ V. parahaemolyticus strains were isolated from shellfish-growing areas in the Pacific Northwest (PNW). We compared clinical and environmental strains isolated from the PNW to those from other geographic regions within the United States and Asia for the presence of virulence-associated genes, including the thermostable direct hemolysin (tdh), the thermostable-related hemolysin (trh), urease (ureR), the pandemic group specific markers orf8 and toxRS, and genes encoding both type 3 secretion systems (T3SS1 and T3SS2). The majority of clinical strains from the PNW were positive for tdh, trh, and ureR genes, while a significant proportion of environmental isolates were tdh⁺ but trh negative. Hierarchical clustering grouped the majority of these clinical isolates into a cluster distinct from that including the pandemic strain RIMD2210633, clinical isolates from other geographical regions, and tdh⁺, trh-negative environmental isolates from the PNW. We detected T3SS2-related genes (T3SS2β) in environmental strains that were tdh and trh negative. The presence of significant concentrations of tdh⁺, trh-negative environmental strains in the PNW that have not been responsible for illness and T3SS2β in tdh- and trh-negative strains emphasizes the diversity in this species and the need to identify additional virulence markers for this bacterium to improve risk assessment tools for the detection of this pathogen.     

Bias in the Listeria monocytogenes Enrichment Procedure: Lineage 2 Strains Outcompete Lineage 1 Strains in University of Vermont Selective Enrichments

Listeria monocytogenes can be isolated from a range of food products and may cause food-borne outbreaks or sporadic cases of listeriosis. L. monocytogenes is divided into three genetic lineages and 13 serotypes. Strains of three serotypes (1/2a, 1/2b, and 4b) are associated with most human cases of listeriosis. Of these, strains of serotypes 1/2b and 4b belong to lineage 1, whereas strains of serotype 1/2a and many other strains isolated from foods belong to lineage 2. L. monocytogenes is isolated from foods by selective enrichment procedures and from patients by nonselective methods. The aim of the present study was to investigate if the selective enrichment procedure results in a true representation of the subtypes of L. monocytogenes present in a sample. Eight L. monocytogenes strains (four lineage 1 strains and four lineage 2 strains) and one Listeria innocua strain grew with identical growth rates in the nonselective medium brain heart infusion (BHI), but differed in their growth rate in the selective medium University of Vermont medium I (UVM I). When coinoculated in UVM I, some strains completely outgrew other strains. This outcome was dependent on the lineage of L. monocytogenes rather than the individual growth rate of the strains. When inoculated at identical cell densities in UVM I, L. innocua outcompeted L. monocytogenes lineage 1 strains but not lineage 2 strains. In addition, lineage 2 L. monocytogenes strains outcompeted lineage 1 L. monocytogenes strains in all combinations tested, indicating a bias in strains selected by the enrichment procedures. Bias also occurred when coinoculating two lineage 2 or lineage 1 strains; however, it did not appear to correlate with origin (clinical versus food). Identical coinoculation experiments in BHI suggested that the selective compounds in UVM I and II influenced this bias. The results of the present study demonstrate that the selective procedures used for isolation of L. monocytogenes may not allow a true representation of the types present in foods. Our results could have a significant impact on epidemiological studies, as lineage 1 strains, which are often isolated from clinical cases of listeriosis, may be suppressed during enrichment by other L. monocytogenes lineages present in a food sample.     

Discovery of a fourth evolutionary lineage of Phytophthora ramorum: EU2

Phytophthora ramorum is a recently introduced, aggressive Phytophthora species that has caused extensive mortality of oak and tanoak trees in the western USA and Japanese larch trees in the UK. P. ramorum is also present on Rhododendron, Camellia, and Viburnum in the nursery industry, which is thought to have been the pathway for its spread into new geographic regions including forests and natural ecosystems. Three lineages of P. ramorum have been described, informally designated EU1, NA1, and NA2, and each lineage is believed to originate from an as yet unknown exotic centre of origin. Preliminary SSR and sequence analysis of isolates from a UK P. ramorum survey revealed seven isolates with profiles that did not match the previously known lineages. Detailed SSR and multilocus sequence analysis of these isolates are presented, allowing us to assign these isolates to a new P. ramorum lineage, designated EU2. Although the known geographical origin of these isolates is currently limited to Northern Ireland and western Scotland, the EU2 lineage isolates have been obtained from four different host plants, including Japanese larch. All isolates are of A1 compatibility type, which implies that this finding does not increase the risk of outcrossing with the EU1 lineage isolates already present in the UK. The oldest EU2 strain was isolated in 2007 but no SSR-based intraEU2 lineage genotypic diversity was detected. The combination of these elements points to a recent introduction, despite emergency phytosanitary measures to control introduction and spread. A PCR-RFLP method for the rapid identification of EU2 lineage isolates is presented.     

Genetic structure and systematic relationships within the Ophrys fuciflora aggregate (Orchidaceae: Orchidinae): high diversity in Kent and a wind-induced discontinuity bisecting the Adriatic

Background and Aims

A recent phylogenetic study based on multiple datasets is used as the framework for a more detailed examination of one of the ten molecularly circumscribed groups identified, the Ophrys fuciflora aggregate. The group is highly morphologically variable, prone to phenotypic convergence, shows low levels of sequence divergence and contains an unusually large proportion of threatened taxa, including the rarest Ophrys species in the UK. The aims of this study were to (a) circumscribe minimum resolvable genetically distinct entities within the O. fuciflora aggregate, and (b) assess the likelihood of gene flow between genetically and geographically distinct entities at the species and population levels.

Methods

Fifty-five accessions sampled in Europe and Asia Minor from the O. fuciflora aggregate were studied using the AFLP genetic fingerprinting technique to evaluate levels of infraspecific and interspecific genetic variation and to assess genetic relationships between UK populations of O. fuciflora s.s. in Kent and in their continental European and Mediterranean counterparts.

Key Results

The two genetically and geographically distinct groups recovered, one located in England and central Europe and one in south-eastern Europe, are incongruent with current species delimitation within the aggregate as a whole and also within O. fuciflora s.s. Genetic diversity is higher in Kent than in the rest of western and central Europe.

Conclusions

Gene flow is more likely to occur between populations in closer geographical proximity than those that are morphologically more similar. Little if any gene flow occurs between populations located in the south-eastern Mediterranean and those dispersed throughout the remainder of the distribution, revealing a genetic discontinuity that runs north–south through the Adriatic. This discontinuity is also evident in other clades of Ophrys and is tentatively attributed to the long-term influence of prevailing winds on the long-distance distribution of pollinia and especially seeds. A cline of gene flow connects populations from Kent and central and southern Europe; these individuals should therefore be considered part of an extensive meta-population. Gene flow is also evident among populations from Kent, which appear to constitute a single metapopulation. They show some evidence of hybridization, and possibly also introgression, with O. apifera.Key words: AFLP, conservation, gene flow, Kent, migration, Ophrys fuciflora, prevailing winds, pseudocopulation, speciation, species delimitation     

New insights on postglacial colonization in western Europe: the phylogeography of the Leisler's bat (Nyctalus leisleri)

Despite recent advances in the understanding of the interplay between a dynamic physical environment and phylogeography in Europe, the origins of contemporary Irish biota remain uncertain. Current thinking is that Ireland was colonized post-glacially from southern European refugia, following the end of the last glacial maximum (LGM), some 20 000 years BP. The Leisler''s bat (Nyctalus leisleri), one of the few native Irish mammal species, is widely distributed throughout Europe but, with the exception of Ireland, is generally rare and considered vulnerable. We investigate the origins and phylogeographic relationships of Irish populations in relation to those across Europe, including the closely related species N. azoreum. We use a combination of approaches, including mitochondrial and nuclear DNA markers, in addition to approximate Bayesian computation and palaeo-climatic species distribution modelling. Molecular analyses revealed two distinct and diverse European mitochondrial DNA lineages, which probably diverged in separate glacial refugia. A western lineage, restricted to Ireland, Britain and the Azores, comprises Irish and British N. leisleri and N. azoreum specimens; an eastern lineage is distributed throughout mainland Europe. Palaeo-climatic projections indicate suitable habitats during the LGM, including known glacial refugia, in addition to potential novel cryptic refugia along the western fringe of Europe. These results may be applicable to populations of many species.