Cumulative temperatures for prediction of the beginning of ash (Fraxinus excelsior L.) pollen season

The aim of this study is the analysis of the meteorological conditions leading to the release of ash pollen in order to predict the beginning of the pollen season in Zurich, Switzerland. For 12 years over a period of 14 consecutive years, it appears that ash pollen release begins when the cumulated mean daily temperatures starting on January 1 reach 220.7°C, admitting a variation of ±30°C that more or less corresponds to 3 consecutive days at 10°C.     

Chloroplast DNA variation and postglacial recolonization of common ash (Fraxinus excelsior L.) in Europe

We used chloroplast polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) and chloroplast microsatellites to assess the structure of genetic variation and postglacial history across the entire natural range of the common ash (Fraxinus excelsior L.), a broad-leaved wind-pollinated and wind-dispersed European forest tree. A low level of polymorphism was observed, with only 12 haplotypes at four polymorphic microsatellites in 201 populations, and two PCR-RFLP haplotypes in a subset of 62 populations. The clear geographical pattern displayed by the five most common haplotypes was in agreement with glacial refugia for ash being located in Iberia, Italy, the eastern Alps and the Balkan Peninsula, as had been suggested from fossil pollen data. A low chloroplast DNA mutation rate, a low effective population size in glacial refugia related to ash's life history traits, as well as features of postglacial expansion were put forward to explain the low level of polymorphism. Differentiation among populations was high (GST= 0.89), reflecting poor mixing among recolonizing lineages. Therefore, the responsible factor for the highly homogeneous genetic pattern previously identified at nuclear microsatellites throughout western and central Europe (Heuertz et al. 2004) must have been efficient postglacial pollen flow. Further comparison of variation patterns at both marker systems revealed that nuclear microsatellites identified complex differentiation patterns in south-eastern Europe which remained undetected with chloroplast microsatellites. The results suggest that data from different markers should be combined in order to capture the most important genetic patterns in a species.     

Frost periods and beginning of the ash (Fraxinus excelsior L.) pollen season in Basel (Switzerland)

In order to be able to forecast the beginning of the ash pollen season, several meteorological factors have been considered. During these research activities it appeared that the ash pollen release is closely linked to frost periods happening during the months of February and March. For 24 years over a period of 29 consecutive years, a daily concentration of 30 ash pollen grains/m³/day has been reached when the cumulated mean daily temperatures starting on the last day when the absolute minimum temperature is lower or equal to −2 °C amounts on average to 214 °C – admitting a variation of plus or minus 42 °C. As for the five remaining years, rain is three times responsible for the delay of the pollen release. This revised version was published online in June 2006 with corrections to the Cover Date.     

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.     

Assessment of genetic structure within and among Bulgarian populations of the common ash (Fraxinus excelsior L.)

We analysed genetic variation within and between populations of the common ash from Bulgaria in order to extract biological information useful in the context of conservation management of eastern European genetic resources of noble hardwood species. A total of 321 trees from three regions of Bulgaria were typed at six highly polymorphic microsatellite loci. Analysis of within-population inbreeding suggests an upper boundary value of 2.7% for the selfing rate. Significant spatial genetic structure consistent with models of isolation by distance was detected within four out of 10 populations as well as among populations. Estimates of neighbourhood size in the range 38-126 individual trees were obtained based on spatial genetic structure analyses at either the intrapopulation or interpopulation level. Differentiation among populations explained only about 8.7% of total genetic diversity. These results are discussed in comparison with data from social broad-leaved species such as oak and beech.     

Underwater germination potential of common ash seed (Fraxinus excelsior L.) originating from flooded and non-flooded sites

Oxygen availability is one of the main factors affecting seed germination capacity. On floodplains, oxygen supply is seasonally restricted because of flooding events. Seed ecotypes are able to cope with total submergence to different extents, depending on the species and its position in a given environmental gradient. This investigation tested three provenances of common ash (Fraxinus excelsior L.), one from a lowland site and two from upland sites, for germination capacity and germination velocity under water. The final germination capacity of all provenances was unaffected, but the germination velocity was decreased for all provenances. The adaptational significance of these results is discussed.     

Diversity of management practices required to ensure conservation of rare and locally threatened plant species in grasslands: a case study at a regional scale (Lorraine, France)

The Lorraine region, in northeast France, harbours 77 protected plantspecies whose main localities are in grasslands. About 80% of them are very rareor rare (less than 15 localities) in that territory and are threatened by changes inagriculture. These species occur in a large range of grassland habitats,reflecting variability of climatic, hydrological and edaphic parameters, butalso of agricultural practices. A more precise analysis of managementrequirements was carried out for seven of these species, Euphorbiapalustris, Aster amellus, Pulsatillaalba, Carex hordeistichos, Thalictrumminus ssp. majus, Botrychiummatricariifolium and Ophioglossum vulgatum, whichwere represented on a triangle whose corners correspond to grazing, cutting andno or very low exploitation. It confirmed the large diversity of managementpractices required to ensure conservation of rare and threatened grassland plantspecies, even at a local level. These appropriate conservation measures havetherefore to be carried out or, at least, monitored by specialized natureconservation organizations.     

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.     

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.     

Shoot initiation in the leaflet axils of compound leaves from micropropagated shoots of juvenile and mature common ash (Fraxinus excelsior L.)

Micropropagation of a mature ash tree has been achieved forthe first time. The main obstacle encountered was contaminationof the initial explants with microorganisms. However, once apparentlysterile shoots had been obtained, shoot proliferation was achievedmost effectively by culturing nodes on Driver and Kuniyuki walnutmedium containing 22.2 µM benzyladenine. After severalsubcultures, a species of Bacillus appeared with the matureculture line, but it did not affect shoot or root developmentadversely. With successive subculturing, shoots of the matureash clone became progressively easier to root. Pinnately compoundleaf explants from micropropagated shoots of two seedlings andthe mature tree, placed on to Murashige and Skoog-based culturemedia supplemented with 4.4µM-phenyl-N-1,2,3-thiadiazol-5-ylurea,produced shoot buds. Most buds developed from the rachis atthe points of attachment of the leaflets. When transferred toDriver and Kuniyuki walnut medium, buds from one of the seedlinglines and from the mature tree, elongated into shoots, and weresubsequently transferred to media for micropropagation and rooting.Shoot initiation is discussed in relation to possible occurrenceof vestigial meristems in the axils of leaflets and the partialshoot theory of leaf structure. Key words: Forestry, leaf structure, partial shoot theory, thidiazuron, tree     

Phenology and life cycle of the annual, <Emphasis Type="Italic">Chamaesyce maculata</Emphasis> (L.) Small (Euphorbiaceae), with multiple overlapping generations in Japan

The phenology of germination, vegetative growth and sexual reproduction in the annual Chamaesyce maculata (L.) Small (Euphorbiaceae) were investigated in a natural population in western Japan. Seedlings emerged from mid-June to early October, with three peaks: mid-June, late July and late August. Plants that emerged in June commenced sexual reproduction from late July, and thereafter both vegetative growth and sexual reproduction occurred together until early November, the plants showing no switching from vegetative growth to sexual reproduction. Seedlings that emerged in June and July suffered high mortality, but most seedlings that emerged from August onward survived until the reproductive stage. The minimum size for reproduction was largest for plants that emerged early in the season, and it decreased with a delay in seedling emergence. The late emergence of seedlings that resulted in low reproductive output may be to some extent compensated for by the increased probability of survival in the seedling stage. A transplant experiment clarified that C. maculata can repeat a maximum of three overlapping generations within a year. Multiple generations per year were attained by non-dormant seeds produced in the first and second generations and clearly resulted in an increased reproductive output per year. The life cycle with multiple overlapping generations may have been acquired in habitats where unpredictable disturbance results in temporally unsuitable conditions for germination, vegetative growth and sexual reproduction of annual plants, but where suitable conditions frequently continue over a period longer than the single generation time of annual plants.     

Nuclear microsatellites reveal contrasting patterns of genetic structure between western and southeastern European populations of the common ash (Fraxinus excelsior L.)

To determine extant patterns of population genetic structure in common ash and gain insight into postglacial recolonization processes, we applied multilocus-based Bayesian approaches to data from 36 European populations genotyped at five nuclear microsatellite loci. We identified two contrasting patterns in terms of population genetic structure: (1) a large area from the British Isles to Lithuania throughout central Europe constituted effectively a single deme, whereas (2) strong genetic differentiation occurred over short distances in Sweden and southeastern Europe. Concomitant geographical variation was observed in estimates of allelic richness and genetic diversity, which were lowest in populations from southeastern Europe, that is, in regions close to putative ice age refuges, but high in western and central Europe, that is, in more recently recolonized areas. We suggest that in southeastern Europe, restricted postglacial gene flow caused by a rapid expansion of refuge populations in a mountainous topography is responsible for the observed strong genetic structure. In contrast, admixture of previously differentiated gene pools and high gene flow at the onset of postglacial recolonization of western and central Europe would have homogenized the genetic structure and raised the levels of genetic diversity above values in the refuges.     

Biotic constraints on the invasion of diffuse knapweed (Centaurea diffusa) in North American grasslands

Knapweeds (Centaurea spp.) are among the most invasive of non-indigenous plant species that have colonized western North America over the last century. We conducted a 4-year experiment in a reconstructed grassland to test hypotheses related to the ability of grasslands to resist the invasion of diffuse knapweed (C. diffusa). We experimentally invaded C. diffusa and three native species into areas where we manipulated soil nitrogen (N) and phosphorus (P) availability and removed extant grasses to reduce competition. We evaluated the growth response of these species to these resources and competitive manipulations. Of the native species that were experimentally added, only one species, Ratibida pinnata (prairie coneflower), established in any numbers. Establishment values in intact vegetation were low for both species, but establishment by C. diffusa (0.02%) clearly outperformed that of R. pinnata (0.001%). Under reduced grass competition, establishment was enhanced, but the values for C. diffusa (0.68%) were not statistically different from those of R. pinnata (0.57%). Neither species performed better under higher soil nutrients in the presence of competing grasses. In plots with both species, biomass of the two planted species was positively correlated, but the biomass of both species was negatively correlated with non-added weedy species. Subsequent harvests of C. diffusa indicated that establishment was enhanced in treatments with higher soil nutrients but that the biomass of these plants could only be enhanced when plant competition was also reduced. These results indicate that C. diffusa can establish in intact grasslands at rates higher than natives, but opportunism rather than competitive ability best describes the invasiveness of C. diffusa. Thus, the mechanisms contributing to the establishment of this knapweed species are different from factors identified as contributing to the dominance of this invader.     

Reproduction and growth of the chaparral geophyte, Zigadenus fremontii (Liliaceae), in relation to fire

Zigadenusfremontii is often a striking component of the flora following firein the chaparral. Like other geophytes, it produces large numbers of flowers inthe first spring after a burn. Although these plants are most conspicuous intheearly postfire environment, the question that remains is, how do they persistinthe interval between fires? To address this we investigated differences in thegrowth and reproduction of Z. fremontii in burned andunburned chaparral. We monitored marked individuals for nine years at threesites: two that were burned in 1990 and one in the same area that was inunburned mature chaparral. We measured leaf area, and production of flowers andfruits. We also conducted seed experiments in the field to determine the ratesand timing of germination. We found that reproduction occurs only in theimmediate postfireperiod: flowering and production of fruits and seeds in the first yearfollowingfire, and seedling establishment by year 3. There was a cost of reproduction;plants that flowered (in the burn area) had negative growth rates the followingyear. In contrast, plants in unburned chaparral, which did not flower, hadpositive growth rates over the same period. Moreover, plants that produced themost flowers had the lowest growth rates. In the unburned chaparral site,plantswere not dormant as predicted from previous literature; instead they producedleaves nearly every year. In most years the average leaf area per plant wasgreater than that in the burned sites. Our results indicate that postfirereproduction depends on growth and carbohydrate storage in the inter-fireperiod. We also suggest that this species is relatively long-lived for aherbaceous perennial.     

Root damage and water stress: treatments affecting the exploitation of the buds of common ash Fraxinus excelsior L., by larvae of the ash bud moth Prays fraxinella Bjerk. (Lep., Yponomeutidae)

Damage to the buds of the common ash (Fraxinus excelsior L.) by larvae of the ash bud moth Prays fraxinella Bjerk. was investigated in trees growing in the presence, or absence, of an adjacent ditch. The presence of a ditch was correlated with increased damage levels due to bud moth. Saplings were used in an experiment to compare the effects of artificial drought and root damage with those of ditches. Both root damage and drought elicited analogous responses to those observed in trees growing near a ditch, being associated with elevated levels of bud moth exploitation and reduced growth of shoots. There exists a documented link between ditches, root disturbance, and the dieback of hedgerow ash trees in Great Britain; the response of P. fraxinella to environmental disturbances associated with dieback is hypothesised to be evidence of the possible role of moth damage as a causal factor in dieback.     

Exploring the use of chamomile (Matricaria chamomilla L.) bioactive compounds to control flixweed (Descurainia sophia L.) in bread wheat (Triticum aestivum L.): Implication for reducing chemical herbicide pollution

Intensive chemical herbicide use has resulted in human health and environmental issues. This study evaluated the phytotoxic potential of chamomile extract as a bioherbicide to minimize chemical herbicide use in wheat production. Treatments including four concentrations (0, 50, 100, and 150 mL/L) of three different chamomile plant parts (root, shoot, and root + shoot) extracts were applied to flixweed as a major weed in wheat production. Except for 50 mL/L of root extract, other concentrations of chamomile extracts decreased the germination rate of flixweed. Germnaiton rate of wheat increased with chamomile extracts except at 150 mL/L concentration of shoot extract at which the germination rate of flixweed and wheat reduced by 71.7 and 35.4%, respectively, compared to respective controls. Compared to wheat, malondialdehyde and proline in flixweed were increased fivefold in flixweed and compared to the control, ranged from 84–473 and 240–1422%, respectively. Chamomile extract also declined cell viability much quicker in flixweed than in wheat reflecting on greater inhibitory effect for flixweed control. Chamomile shoot extract reduced seedling weight and vigor index of flixweed by 63.75 and 59.4%, respectively, compared to the respective control. Results of liquid chromatography mass spectrometry of chamomile extract indicated polyphenols, flavonoids, terpenoids, and bioactive phenolic coumarins, glycosylated derivatives, quercetin and its derivatives, herniarin, umbelliferone, P-cymene, chamazulene, farnesol, amitrole, 1,8-cineole, and limonene were effective in inhibiting the germination and growth of flixweed. We concluded that 150 mg/L of chamomile shoot extract could be used as a bioherbicide to sustainably suppress flixweed in wheat production.