Micromorphological differences between some European and American <Emphasis Type="Italic">Fraxinus</Emphasis> (<Emphasis Type="Italic">Oleaceae</Emphasis>) species

Micromorphological differences in leaves and pollen between two American (Fraxinus americana L., F. pennsylvanica Marshall) and two European (F. angustifolia Vahl, F. excelsior L.) ash species were studied using scanning electron microscope. The types, dimensions and distribution of characteristic trichomes were established and measured. Capitate hairs on the leaves had the same shape in all researched ash species. Acicular hairs were regularly present in two American ash species, but very rarely in the glabrous phase of F. angustifolia and F. excelsior. Only F. americana had coronulate abaxial surface of leaves. Pollen of F. angustifolia and F. excelsior had 3 (tricolpate) apertures, and F. americana and F. pennsylvanica 4 (stephanocolpate) apertures. Based on the appearance of the reticulum it’s possible to clearly distinguish all four species. F. angustifolia and F. pennsylvanica had muri with transversal ridges and seldom granules. Muri of F. excelsior and F. americana had slightly visible transversal ridges, and because of that noticeable granules.     

Thank you for not flowering: conservation genetics and gene flow analysis of native and non-native populations of Fraxinus (Oleaceae) in Ireland

The risks of gene flow between interfertile native and introduced plant populations are greatest when there is no spatial isolation of pollen clouds and phenological patterns overlap completely. Moreover, invasion probabilities are further increased if introduced populations are capable of producing seeds by selfing. Here we investigated the mating system and patterns of pollen-mediated gene flow among populations of native ash (Fraxinus excelsior) and mixed plantations of non-native ash (F. angustifolia and F. excelsior) as well as hybrid ash (F. excelsior × F. angustifolia) in Ireland. We analysed the flowering phenology of the mother trees and genotyped with six microsatellite loci in progeny arrays from 132 native and plantation trees (1493 seeds) and 444 potential parents. Paternity analyses suggested that plantation and native trees were pollinated by both native and introduced trees. No signs of significant selfing in the introduced trees were observed and no evidence of higher male reproductive success was found for introduced trees compared with native ones either. A small but significant genetic structure was found (φ_ft=0.05) and did not correspond to an isolation-by-distance pattern. However, we observed a significant temporal genetic structure related to the different phenological groups, especially with early and late flowering native trees; each phenological group was pollinated with distinctive pollen sources. Implications of these results are discussed in relation to the conservation and invasiveness of ash and the spread of resistance genes against pathogens such as the fungus Chalara fraxinea that is destroying common ash forests in Europe.     

Organization of nuclear ribosomal DNA and species-specific polymorphism in closely related Fraxinus excelsior and F. oxyphylla

The ribosomal DNA repeat units of two closely related species of the genus Fraxinus, F. excelsior and F. oxyphylla, were characterized. The physical maps were constructed from DNA digested with BamHI, EcoRI, EcoRV and SacI, and hybridized with three heterologous probes. The presence or the absence of an EcoRV restriction site in the 18s RNA gene characterizes two ribosomal DNA unit types found in both species and which coexist in all individuals. A third unit type appeared unique to all individuals of F. oxyphylla. It carries an EcoRI site in the intergenic spacer. Each type of unit displayed length variations. The rDNA unit length of F. excelsior and F. oxyphylla was determined with EcoRV restriction. It varied between 11kb and 14.5kb in F. excelsior and between 11.8kb to 13.8kb in F. oxyphylla. Using SacI restriction, at least ten spacer length variants were observed in F. excelsior, for which a detailed analysis was conducted. Each individual carries 2–4 length variants which vary by a 0.3-kb step multiple. This length variation was assigned to the intergenic spacer. By using the entire rDNA unit of flax as probe in combination with EcoRI restriction, each species can be unambiguously discriminated. The species-specific banding pattern was used to compare trees from a zone of sympatry between the two species. In some cases, a conflicting classification was obtained from morphological analysis and the use of the species-specific rDNA polymorphism. Implications for the genetic management of both species are discussed.     

Assortative mating and differential male mating success in an ash hybrid zone population

Background  

The structure and evolution of hybrid zones depend mainly on the relative importance of dispersal and local adaptation, and on the strength of assortative mating. Here, we study the influence of dispersal, temporal isolation, variability in phenotypic traits and parasite attacks on the male mating success of two parental species and hybrids by real-time pollen flow analysis. We focus on a hybrid zone population between the two closely related ash species Fraxinus excelsior L. (common ash) and F. angustifolia Vahl (narrow-leaved ash), which is composed of individuals of the two species and several hybrid types. This population is structured by flowering time: the F. excelsior individuals flower later than the F. angustifolia individuals, and the hybrid types flower in-between. Hybrids are scattered throughout the population, suggesting favorable conditions for their local adaptation. We estimate jointly the best-fitting dispersal kernel, the differences in male fecundity due to variation in phenotypic traits and level of parasite attack, and the strength of assortative mating due to differences in flowering phenology. In addition, we assess the effect of accounting for genotyping error on these estimations.     

Relationships between flowering development of Ulmus pumila and Fraxinus excelsior and their airborne pollen

The flowering patterns of Ulmus pumila and Fraxinus excelsior were studied during the 1990 growing season in order to investigate their pollen dispersion curves. The use of airborne pollen records as predictors of flowering of allergenic trees was evaluated. A sampling method to describe quantitative flowering phenophases was applied. A Burkard trap recorded airborne pollen in the city of Mar del Plata. The phenological sampling method showed that floral phenophase development is not always synchronous between trees. The variability registered was larger for F. excelsior than for U. pumila. A delay between flowering and the airborne pollen recorded was not noticed for Ulmus. The greatest amount of Ulmus pollen came from the local vegetation, mainly from U. pumila. Fraxinus pollen was recorded for a longer period after the F. excelsior flowering season the ended. Fraxinus pollen income came from F. excelsior but also from F. americana and F. excelsior var. aurea that flower later. An immediate decrease in airborne concentration was noticed for both pollen types following rain. When relatively strong rainfall was registered a decrease in the number of open flowers occurred only in F. excelsior.     

Effects of the 2003 heatwave and climatic warming on mollusc communities of the Saône: a large lowland river and of its two main tributaries (France)

The study of the dynamics of mollusc populations of the Saône and its two main tributaries, the Doubs and Ognon, over several years has provided us with the opportunity of highlighting the consequences of climatic warming and especially of the heatwave of 2003 on these organisms. From 1987 to 2003, the mean temperature of the waters of the Saône upstream of Lyon (Couzon) increased by 1.5°C. In addition, the summer of 2003 was the hottest since 1500 at least. We used correspondence analysis to identify structure change in mollusc data dating from September 1996 to December 2004. The results revealed: (1) during the period from September 1996 to July 2003, a significant progressive change in the mollusc community structure of the Saône upstream of Lyon, probably linked to the increase of temperature; (2) from July to August 2003 during the heatwave, a sudden change in the structure of mollusc communities and a significant decrease of species richness and density of gastropods and bivalves. During 2004, mollusc density and particularly that of Pisidium remained dramatically low. Similar observations were performed at four other sites along the Saône and in the lower reaches of its two main tributaries. This suggests that the resilience of the mollusc populations (i.e., the speed with which they return to a predisturbance state) to the heatwave is low. In this way, as different climatic models have predicted an increase in the frequency of summers as hot as that of 2003 during this century, more than half the mollusc species currently inhabiting the potamic area of the Saône, Doubs and Ognon, and probably other large rivers, are probably directly threatened with extinction.     

Seasonal pattern of mannitol and malate accumulation in leaves of two manna ash species (Fraxinus ornus L. and F. angustifolia Vahl) growing in Sicily

ABSTRACT

The content of mannitol and malate was assayed enzimatically during spring, summer and autumn, in leaves of two species of ash, Fraxinus ornus L. and Fraxinus angustifolia Vahl, traditionally cultivated in Sicily for the extraction of manna. Both species contain high levels of mannitol and show, on a dry weight basis, a 65–80% increase in the summer content of this polyol. The malate content differs in the two species: in F. ornus it shows a summer increase, but it is relatively low (65–115 µmol g^-1 DW), while in F. angustifolia it is higher (275–318 µmol g^-1 DW), but remains more or less constant throughout the year. The results suggest that in these species, under the local field conditions, mannitol has a more relevant role than malate in the response to summer drought.     

Molecular biodiversity and population structure in common ash (Fraxinus excelsior L.) in Britain: implications for conservation

Current forestry policy promotes the use of local seed for new plantings, on the assumption that local material may be better adapted to local conditions. However, landscape‐scale genetic studies which are necessary to underpin conservation and breeding strategies are often lacking. We investigated molecular diversity in common ash (Fraxinus excelsior L.) sampled from 42 British and six French sites with microsatellites. Chloroplast haplotype H04 was the most common and widespread in Britain, although rare and localized individuals with H02 and H09 were also detected. In addition, three new chloroplast haplotypes were identified, and these were rare and highly localized. In terms of nuclear microsatellite markers, allelic richness differed between sites and decreased in an east to west direction. Differentiation between sites was often very low (mean F_ST 0.025), indicating few differences between the majority of sites. There was a clear excess of homozygotes (mean H_O 0.669, mean H_E 0.818) and a relatively high F_IS (mean 0.182), suggests a consistent level of inbreeding or a widespread Wahlund effect in many F. excelsior sites. Gene pool ancestry analysis suggested that the majority of British F. excelsior belongs to a single meta‐population which covers mainland western and central Europe. Three northern and western sites diverged markedly from the dominant population, and may represent remnants of two late potential Ice Age refugia in northern Britain. The data provide new information which will aid development of appropriate conservation policies for ash and other wind pollinated tree species.     

A new set of mono‐ and dinucleotide chloroplast microsatellites in Fagaceae

A new set of 14 chloroplast microsatellites, represented by mono‐ and dinucleotide repeats, was optimized in the three main species of the Fagaceae (Castanea sativa, Fagus sylvatica and Quercus petraea). The intraspecific variation was tested in some natural populations. The polymorphic microsatellites displayed two or three variants. Conservation of the primer pairs was checked on an additional set of six species of the Fagaceae and on Fraxinus excelsior. All the primer pairs produced a fragment of the expected size in the Fagaceae species while no amplification was obtained with 36% of the primers in F. excelsior.     

Molecular–cytogenetic studies of ribosomal RNA genes and heterochromatin in three European Fraxinus species

Three European representatives of the genus Fraxinus were studied for the first time for their rDNA and heterochromatin patterns. The physical mapping of two rRNA gene families 5S and 18S–5.8S–26S (45S) and the distributional pattern of GC-rich regions in the chromosomes have been established by means of fluorescence in situ hybridization (FISH) and fluorochrome banding with chromomycin A₃. The genome size was assessed by flow cytometry. Heterochromatin and rDNA organization was conserved and almost identical for two species from Fraxinus section (F. angustifolia and F. excelsior). The number and position of rDNA loci in F. ornus (section Ornus) were similar; however, the organization of genes was quite different. In this species the 5S and 45S rRNA genes were colocalized at the level of satellites of two chromosome pairs bearing nucleolar organizing regions (NORs). One 5S locus was also observed under the 45S one of one chromosome pair. In F. angustifolia and F. excelsior, only 45S loci were situated at the level of satellites and secondary constrictions, while 5S was located just under 45S in the distal part of the short arm of one out of two marked pairs. The number and position of GC-rich DNA correspond to those of 45S loci. The genome size (2C value) was of 1.54 and 1.68 pg for F. angustifolia and F. excelsior, respectively. Fraxinus ornus possessed the highest 2C DNA value (1.98 pg). In the light of these cytogenetic features the clear differentiation between two sections (Fraxinus and Ornus) was observed both at the rDNA and genome size levels.     

Defining the twig fungal communities of Fraxinus species and Fraxinus excelsior genotypes with differences in susceptibility to ash dieback

Ash dieback disease (caused by Hymenoscyphus fraxineus) has affected European ash species (Fraxinus spp.) in recent decades. However, some Asian and American species of Fraxinus and certain genotypes of Fraxinus excelsior are less affected by the disease. We used ITS1-metabacoding to explore the drivers influencing diversity and composition of the twig fungal communities of Fraxinus species and F. excelsior genotypes. Our results revealed that fungi in the classes Eurotiomycetes and Dothideomycetes were among the most prevalent taxa in both Fraxinus species and F. excelsior genotypes. The diversity of the fungal communities differed significantly among Fraxinus species and could be explained by seed origin. Neither host genotype nor season had a significant effect on the community diversity of F. excelsior genotypes. On the other hand, the composition of twig fungal communities differed significantly among host species and among F. excelsior genotypes, and in F. excelsior there was also a significant effect of season on the composition of the fungal community. We did not find a clear effect of ash dieback susceptibility on either diversity or composition of fungal communities in twigs of Fraxinus species, although the effect was significant on the composition of fungal communities among F. excelsior genotypes. Our results demonstrated differences in fungal communities among species of Fraxinus and of F. excelsior genotypes, suggesting specific relationship between individual host genotypes and endophytic fungi.     

Cloning and analysis of a 6.8-kb rDNA intergenic spacer region of the European ash (Fraxinus excelsior L.)

The 6.8-kb rDNA intergenic spacer region of F. excelsior was isolated from a CsCl/actinomycin-D gradient and cloned into pUC18 for further characterization. We observed the presence of subrepeats delimited by HaeIII enzyme sites. These subrepeats were sub-cloned and 11 clones were sequenced. These corresponded to subrepeated elements of either 32 bp or 41 bp that shared a 23-bp common sequence in the 5 end. Within each family of subrepeats, the percentage of common nucleotides was 84.4% for the 5 32-bp subrepeats and 67.4% for the 640-bp subrepeats. Non-repeated HaeIII fragments of 450 bp and 650 bp were also sub-cloned. To compare homology at the IGS region between the rDNA spacers of F. excelsior and the three related species (F. oxyphylla, F. americana, F. ornus), we conducted Southern hybridization analyses using each member of the 32-bp and 40-bp subrepeat families and the unique 450-bp and 650-bp fragments as probes. These analyses indicated that (1) the American ash is more genetically distant from the other three species that the latter are from each other and (2) F. oxyphylla and F. excelsior are more closely related to each other than to F. ornus.     

Estimating Coextinction Risks from Epidemic Tree Death: Affiliate Lichen Communities among Diseased Host Tree Populations of Fraxinus excelsior

At least 10% of the world’s tree species are threatened with extinction and pathogens are increasingly implicated in tree threats. Coextinction and threats to affiliates as a consequence of the loss or decline of their host trees is a poorly understood phenomenon. Ash dieback is an emerging infectious disease causing severe dieback of common ash Fraxinus excelsior throughout Europe. We utilized available empirical data on affiliate epiphytic lichen diversity (174 species and 17,800 observations) among 20 ash dieback infected host tree populations of F. excelsior on the island Gotland in the Baltic Sea, Sweden. From this, we used structured scenario projections scaled with empirical data of ash dieback disease to generate probabilistic models for estimating local and regional lichen coextinction risks. Average coextinction probabilities (Ā) were 0.38 (95% CI ±0.09) for lichens occurring on F. excelsior and 0.14 (95% CI ±0.03) when considering lichen persistence on all tree species. Ā was strongly linked to local disease incidence levels and generally increasing with lichen host specificity to F. excelsior and decreasing population size. Coextinctions reduced affiliate community viability, with significant local reductions in species richness and shifts in lichen species composition. Affiliates were projected to become locally extirpated before their hosts, illuminating the need to also consider host tree declines. Traditionally managed open wooded meadows had the highest incidence of ash dieback disease and significantly higher proportions of affiliate species projected to go extinct, compared with unmanaged closed forests and semi-open grazed sites. Most cothreatened species were not previously red-listed, which suggest that tree epidemics cause many unforeseen threats to species. Our analysis shows that epidemic tree deaths represent an insidious, mostly overlooked, threat to sessile affiliate communities in forested environments. Current conservation and management strategies must account for secondary extinctions associated with epidemic tree death.     

Nitrate reduction and nitrate content in ash trees (Fraxinus excelsior L.): distribution between compartments,site comparison and seasonal variation

Summary Nitrate reductase activity (NRA), nitrate content and biomass components of leaflets, leaf stalks, old stem, current-year stem and roots of ash trees (Fraxinus excelsior L.) growing in their natural habitats were investigated. In addition, NRA, total nitrogen and nitrate concentration were analyzed in the leaves and roots of ash trees from four different field sites. The highest NRA per gram biomass and also per total compartment biomass was found in the leaflets, even though root biomass was much higher than total leaflet biomass. The highest nitrate concentrations were found in the leaf stalks. Correlations between nitrate availability in the soil and NRA in leaves were not significant due to high variability of the actual soil nitrate concentrations. The seasonal variation in foliar NRA, nitrate concentration and total nitrogen concentration is much smaller in F. excelsior than reported for herbaceous species and is mainly caused by changes in the actual soil nitrate availability and by senescence of the leaves.     

Success in the demographic expansion of Fraxinus excelsior L.

This review examines the ecological traits and ecophysiological characteristics of the common ash (Fraxinus excelsior L.) that help to explain the current spread of ash communities in Europe. As a consequence of anthropogenic disturbance, the common ash has experienced new environmental conditions and the frequency of the species has fluctuated over space and time. Owing to its ubiquity, over the last few decades the common ash has been greatly favoured in areas abandoned by agriculture and also along impounded rivers. F. excelsior is a mesophilic species that usually thrives on alluvial soils and can also survive the strong water deficit on hillslopes. Drought tolerance in ash trees is related to stomatal regulation, a decrease in osmotic potential and also an increase in the elastic modulus ). Malic acid plays a central role in drought tolerance, first as one of the major solutes involved in osmotic adjustment, and second as an effector for stomatal regulation through a drought-induced increase in its xylem concentration.     

THE OCCURRENCE OF ADVENTITIOUS AND PREVENTITIOUS BUDS WITHIN THE BARK OF SOME TEMPERATE AND TROPICAL TREES

Endogenous adventitious buds develop in situ from dedifferentiated parenchyma cells of the trunk-bark in the temperate trees Tilia platyphyllos, Acer pseudoplatanus, and Fraxinus excelsior, as well as in the tropical cauliflorous trees Artocarpus integrifolia, Swartzia schomburgkii, and Couroupita guianensis. On aerial roots of Clusia rosea endogenous adventitious buds originate within the proliferated phelloderm beneath lenticels. In Salix alba, Fraxinus excelsior, and Terminalia arjuna, exogenous dormant buds are overgrown during secondary growth and engulfed within the bark tissue, so that they give the impression of apparent endogeny. In the leaf axils of young shoots of Araucaria angustifolia, superficial tissue layers divide and form axillary protrusions, which soon become parenchymatic and partly suberized. A few cells at their bases stay meristematic, however, and develop as rudimentary endogenous bud primordia, which persist in the bark for many years.     

Characterization and primer development for amplification of chloroplast microsatellite regions of Fraxinus excelsior

This study reports the characterization of chloroplast DNA (cpDNA) variation of Fraxinus excelsior at five loci and the successful development of primer pairs for the amplification of three of these containing mononucleotide microsatellites. We detected high levels of haplotype variation among provenances of Fraxinus around Europe and within Ireland.     

The response of life‐history traits to a new species in the community: a story of Drosophila parasitoids from the Rhône and Saône valleys

In the present study, we investigated the evolution of life‐history traits in the main species of a community, after the arrival of a new competitor. Two parasitoid species, Leptopilina heterotoma and Asobara tabida, are present throughout the Rhône and Saône valleys, whereas a third species, Leptopilina boulardi, is slowly extending its distribution northwards. In the presence of L. boulardi, competing parasitoids experience a higher mortality and lower host availability. Resources should thus be re‐allocated between traits according to these new factors. We compared life‐history traits of populations of L. heterotoma and A. tabida in areas with and without L. boulardi. As predicted by both Price's balanced mortality hypothesis and the theory of life‐history traits, we found that investment in reproduction is higher in southern populations for both native species, coupled with higher travelling abilities. However, only A. tabida paid their higher fecundity by a lower longevity. The absence of a clear trade‐off between these traits in L. heterotoma may be explained by a lower metabolic rate in southern populations. These results highlight the importance of the community change over climate in the evolution of life‐history traits in this parasitoid community. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ?? , ??–??.     

Spatio-temporal dynamics of endophyte diversity in the canopy of European ash (Fraxinus excelsior)

Leaf-inhabiting endophytic fungi of Fraxinus excelsior growing in a floodplain forest were isolated during 2008 to investigate vertical community structure, species richness and seasonal variation. The analysis of 848 fungal endophytes from 213 leaves resulted in 50 different species. In the understorey, infection density and species richness were higher than in the crowns of mature trees throughout the whole vegetation period. Within tree crowns, sun-exposed leaves of the top canopy exhibited the lowest infection rates. Most species were rare or absent in spring and in the light crowns and frequent in autumn and the understorey. However, some species, especially the two most frequent, Alternaria infectoria and A. alternata, deviated from these patterns. Young leaves were nearly free of endophytes. Apparently, the subsequent infection and establishment of fungi strongly depend on microclimatic parameters and leaf characters, which create highly variable spatial and temporal colonisation patterns within an individual tree.     

Presence of natural genetic resistance in Fraxinus excelsior (Oleraceae) to Chalara fraxinea (Ascomycota): an emerging infectious disease

Fraxinus excelsior, common ash native to Europe, is threatened by a recently identified pathogenic fungus Chalara fraxinea, which causes extensive damage on ash trees across Europe. In Denmark, most stands are severely affected leaving many trees with dead crowns. However, single trees show notably fewer symptoms. In this study, the impact of the emerging infectious disease on native Danish ash trees is assessed by estimating presence of inherent resistance in natural populations. Disease symptoms were assessed from 2007 to 2009 at two different sites with grafted ramets of 39 selected clones representing native F. excelsior trees. A strong genetic variation in susceptibility to C. fraxinea infections was observed. No genetic or geographic structure can explain the differences, but strong genetic correlations to leaf senescence were observed. The results suggest that a small fraction of trees in the Danish population of ash possess substantial resistance against the damage. Though this fraction is probably too low to avoid population collapse in most natural or managed ash forests, the observed presence of putative resistance against the emerging infectious disease in natural stands is likely to be of evolutionary importance. This provides prospects of future maintenance of the species through natural or artificial selection in favour of remaining healthy individuals.