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Genetic differentiation of continental and island populations of Bombus terrestris (Hymenoptera: Apidae) in Europe.

Ten microsatellite loci and a partial sequence of the COII mitochondrial gene were used to investigate genetic differentiation in B. terrestris, a bumble bee of interest for its high-value crop pollination. The analysis included eight populations from the European continent, five from Mediterranean islands (six subspecies altogether) and one from Tenerife (initially described as a colour form of B. terrestris but recently considered as a separate species, B. canariensis). Eight of the 10 microsatellite loci displayed high levels of polymorphism in most populations. In B. terrestris populations, the total number of alleles detected per polymorphic locus ranged from 3 to 16, with observed allelic diversity from 3.8 +/- 0.5 to 6.5 +/- 1.4 and average calculated heterozygosities from 0.41 +/- 0.09 to 0.65 +/- 0.07. B. canariensis showed a significantly lower average calculated heterozygosity (0.12 +/- 0.08) and observed allelic diversity (1.5 +/- 0.04) as compared to both continental and island populations of B. terrestris. No significant differentiation was found among populations of B. terrestris from the European continent. In contrast, island populations were all significantly and most of them strongly differentiated from continental populations. B. terrestris mitochondrial DNA is characterized by a low nucleotide diversity: 0.18% +/- 0.07%, 0.20% +/- 0.04% and 0.27% +/- 0.04% for the continental populations, the island populations and all populations together, respectively. The only haplotype found in the Tenerife population differs by a single nucleotide substitution from the most common continental haplotype of B. terrestris. This situation, identical to that of Tyrrhenian islands populations and quite different from that of B. lucorum (15 substitutions between terrestris and lucorum mtDNA) casts doubts on the species status of B. canariensis. The large genetic distance between the Tenerife and B. terrestris populations estimated from microsatellite data result, most probably, from a severe bottleneck in the Canary island population. Microsatellite and mitochondrial DNA data call for the protection of the island populations of B. terrestris against importation of bumble bees of foreign origin which are used as crop pollinators.

Alleles↗

Inbreeding and population structure in two pairs of cryptic fig wasp species.

We used recently developed microsatellites to directly estimate inbreeding levels in two pairs of coexisting cryptic fig wasp species ('Pegoscapus hoffmeyeri sp. A and sp. B', 'P. gemellus sp. A and sp. B'). Previous tests of Hamilton's local mate competition (LMC) theory in fig wasps have used the number of dead foundresses in a fig fruit to indirectly estimate the relative contribution of each to the common brood and thereby the level of local mate competition. Further, the population level of inbreeding has been indirectly estimated using the distribution of foundress numbers across broods. Our direct genetic estimates confirmed previous assumptions that the species characterized by lower foundress numbers showed higher relative levels of inbreeding. However, there were quantitative differences between the observed level of inbreeding and the expectation based on the distribution of foundress numbers in both pollinator species associated with Ficus obtusifolia. Here, genotype compositions of broods revealed that only 23% of fruits with multiple foundresses actually contained brood from more than one foundress, thus explaining at least part of the underestimate of actual sibmating. Within the four wasp species there was no evidence for genetic differentiation among the wasp populations sampled from different trees across 20 km and from different points in time. Further, no genotypic disequilibrium was detected within any of the species. Although F1 hybrids were observed between the two species pollinating F. obtusifolia, there was no evidence of genetic introgression. Finally, we found that 11% of the sons of allospecifically mated mothers were diploid hybrids suggesting a break down of the sex determination system in hybrids.

Animals↗

Increased selfing and correlated paternity in a small population of a predominantly outcrossing conifer, Pinus sylvestris.

Outcrossing rate, the rates of ovule and seed abortion, and levels of correlated paternity were estimated in a small population of Pinus sylvestris, a predominantly outcrossing conifer, and were compared with estimates from two widely dispersed woodlands of the same species, showing a range of densities. On average, seed trees of the small population showed an eight-fold higher selfing rate (25 vs. 3%) and a 100-fold greater incidence of correlated paternity (19.6 vs. 0.2%) than did trees from the large populations. No evidence was found of pollen limitation within the remnant stand, as suggested by ovule abortion rates. Investigation of the mating patterns in the small population, based on the unambiguous genealogy of 778 open-pollinated seeds, showed a large departure from random mating. Only 8% of the possible mating pairs within the stand were observed. Correlated paternity rate within a maternal sibship was negatively associated (rs = -0.398, P < 0.050) with the distance to the nearest neighbour, and shared paternity among maternal sibships was negatively correlated (rs = -0.704, P < 0.001) with the distance between seed trees. Numerical simulations, based on the estimated individual pollen dispersal kernel, suggest that restricted dispersal might have been the key factor affecting mating patterns in the small population and, together with low population density, may account for the observed mating system variation between the small and the large populations. The results of this study show that a severe size reduction may substantially affect the mating system of a wind-pollinated, typically outcrossed plant species.

Computer Simulation↗

Spatial genetic structure within a metallicolous population of Arabidopsis halleri, a clonal, self-incompatible and heavy-metal-tolerant species.

Arabidopsis halleri, a close wild relative of A. thaliana, is a clonal, insect-pollinated herb tolerant to heavy metals (Zn, Pd, Cd) and a hyperaccumulator of Zn and Cd. It is of particular interest in the study of evolutionary processes and phytoremediation. However, little is known about its population gene flow patterns and the structure of its genetic diversity. We used five microsatellite loci to investigate the genetic structure at a fine spatial scale (10 cm to 500 m) in a metallicolous population of A. halleri. We also studied the contributions made by clonal propagation and sexual reproduction (seed and pollen dispersal) to the genetic patterns. Clonal diversity was high (D(G) > 0.9). Clonal spread occurs only at short distances (< 1 m). Both clonal spread and limited dispersal, associated with sexual reproduction, contribute to the significant spatial genetic structure revealed by spatial autocorrelation analysis. The shape of the autocorrelogram suggests that seed dispersal is restricted and pollen flow extensive, which may be related to intense activity by insect pollinators. Clonal spread was more extensive in the lowly polluted zone than in the highly polluted zone. This cannot be interpreted as a strategy for promoting the propagation of adapted genotypes under the harshest ecological constraints (highest heavy metal concentrations). The higher fine-scale spatial genetic structure found in the lowly polluted zone can be ascribed to plant densities that were lower than in the highly polluted zone. No evidence of genetic divergence due to spatial heavy metal heterogeneity was found between lowly and highly polluted zones.

Analysis of Variance↗

Interannual genetic heterogeneity of pollen pools accepted by Quercus salicina individuals.

Since flowering often varies among years in wind-pollinated woody species, the genetic composition of pollen pools accepted by seed parents can differ between years. The interannual heterogeneity of pollen flow may be important for maintaining genetic diversity within populations because it can increase genetic variation within populations and the effective sizes of the populations. In this study we examined heterogeneity, using paternity analysis and analysis of molecular variance, in the genetic composition of pollen pools among different reproductive years for six Quercus salicina seed parents in an 11.56-ha plot in a temperate old-growth evergreen broadleaved forest. The genotypes at seven microsatellite loci were determined for 111 adult trees and 777 offspring of the six seed parents in 2-5 reproductive years. Genetic differentiation of pollen pools among different reproductive years for each seed parent was significant over all seed parents and for each of four seed parents that were analysed for more than 2 years, but not for either of the other two seed parents (analysed for 2 years). For both the pollen pools originating from inside the plot and those originating from outside it, genetic differentiation among different reproductive years for each seed parent was significant over all seed parents. However, among-year genetic differentiation in the pollen pools originating from within the plot was detected for all four of the seed parents that were analysed for more than 2 years, but for only one of the four in the pools originating from outside the plot. Genetic diversity (estimated as allelic richness and gene diversity) was higher for pollen pools over all reproductive years than for pollen pools in single years. These results indicate that the year-to-year genetic variation of pollen pools increases genetic diversity in offspring and is strongly affected by the variation in pollen parents within the plot because of their high pollination contributions. The high year-to-year variation in pollen parents within the plot and overall supports the hypothesis that the offspring produced across years represent a larger genetic neighbourhood.

Analysis of Variance↗

A new seed-based assay for meiotic recombination in Arabidopsis thaliana.

Meiotic recombination is a fundamental biological process that plays a central role in the evolution and breeding of plants. We have developed a new seed-based assay for meiotic recombination in Arabidopsis. The assay is based on the transformation of green and red fluorescent markers expressed under a seed-specific promoter. A total of 74 T-DNA markers were isolated, sequenced and mapped both physically and genetically. Lines containing red and green markers that map 1-20 cM apart were crossed to produce tester lines with the two markers linked in cis yielding seeds that fluoresced both in red and green. We show that these lines can be used for efficient scoring of recombinant types (red only or green only fluorescing seeds) in a seed population derived from a test cross (backcross) or self-pollination. Two tester lines that were characterized during several generations of backcross and self-pollination, one in the background of ecotype Landsberg and one in the ecotype Columbia, are described. We discuss the number of plants and seeds to be scored in order to obtain reliable and reproducible crossing over rate values. This assay offers a relatively high-throughput method, with the benefit of seed markers (similar to the maize classical genetic markers) combined with the advantages of Arabidopsis. It advances the prospect to better understand the factors that affect the rate of meiotic crossover in plants and to stimulate this process for more efficient breeding and mapping.

Arabidopsis↗

Investigation of the involvement of Echium plantagineum (Paterson's curse) in seasonal allergy. IgE antibodies to Echium and other weed pollens.

The possible allergenicity of an insect pollinated weed, Echium plantagineum, was investigated in a rural area of Australia. Sixty-one subjects with respiratory allergy were studies. Positive skin test reactions to defatted ammonium bicarbonate extract of pollen were found in over 60% of subjects, and positive RAST tests in a similar number. The question of crossreactivity between weed pollens is discussed. The pollen of E. plantagineum was shown to reach the atmosphere in significant amounts about 1 month before the peak grass pollinating period. Evidence that the pollen of E. plantagineum becomes airborne and elicits an IgE response suggests that further attention should be directed to weed pollens as potential allergens.

Adolescent↗

Thigmomorphogenesis: anatomical, morphological and mechanical analysis of genetically different sibs of Pinus taeda in response to mechanical perturbation.

Twenty-three open pollinated families (half-sibs) and four controlled pollinated families (full-sibs) of Pinus taeda L. (loblolly pine) were grown in a greenhouse and analyzed for changes induced by mechanical perturbation (MP). These changes included inhibition of stem and needle elongation, bracing of branch nodes, and increased radial growth in the direction of the MP. Inhibition of stem elongation was the least variable feature measured. Leaf extension and stem diameter were highly variable between half-sibs. MP induced increased drag in greenhouse grown P. taeda in wind-tunnel tests. In P. taeda, MP induced decreased flexibility and increased elasticity and plasticity of the stem. The increased radial growth of the stems overrode the increase in elasticity, resulting in an overall decrease in flexibility. MP trees had a higher rupture point than non-MP controls. Increased radial growth is a result of more rapid cell divisions of the vascular cambium, resulting in increased numbers of tracheids. The decreased leader growth is partly due to a decreased tracheid length in response to MP.

Biomechanical Phenomena↗

Trade-offs between sexual and clonal reproduction in an aquatic plant: experimental manipulations vs. phenotypic correlations.

That trade-offs result from the allocation of limited resources is a central concept of life history evolution. We quantified trade-offs between sexual and clonal reproduction in the aquatic plant, Butomus umbellatus, by experimentally manipulating sexual investment in two distinct nutrient environments. Increasing seed production caused a significant but nonlinear trade-off. Pollinating half of all flowers strongly reduced clonal bulbil production, but pollinating the remaining flowers did not cause any further trade-off. Trade-offs were not stronger under low nutrient conditions that clearly limited plant growth. Experimentally induced trade-offs were not reflected in negative phenotypic correlations between sexual and clonal allocation among plants within eight populations grown in a uniform greenhouse environment. Diminishing effects of increased sexual allocation plus a lack of accord between experimental manipulations and phenotypic correlations suggest that trade-offs between sexual and clonal reproduction are unlikely to constrain the evolution of reproductive strategy in this species.

Analysis of Variance↗

Storage proteins in Zea mays (L.): interrelationship of albumins, globulins and zeins in the opaque-2 mutation.

Mature endosperms from opaque-2 mutant seeds with various genetic backgrounds (B 37, W 64A, R 802A and Oh 43) contained twice as much globulin than the corresponding normals, reduced amounts of zein and an increased amount of albumin. The latter is caused by a diminished disappearance of albumins in mutant endosperms during the final phases of development as compared to normal endosperms. Albumins from the opaque-2 endosperms appeared on gel electrophoresis as a similar but heterogenous polypeptide population comparable to that of normal endosperms except that in all opaque-2 forms a 70-kDa polypeptide was increased. Similarly, one to three specific globulin polypeptides (47 kDa, 52 kDa and 58 kDa) were, depending on the genotype, increased in the mutant lines. The accumulation of albumins, globulins and zeins, studied in developing W 64A opaque-2 mutant and corresponding normal maize kernels from 10 days after pollination until maturity, demonstrated two-phasic accumulation patterns for all proteins in the mutant, the first phase extending to about 30 days post pollination and the second one from there on until maturity. The transition time-point from first to second phase was characterized in mutant endosperm by a sudden reduction in accumulating albumins (also seen in normal endosperm), by an enhanced accumulation of globulins and the cessation of further accumulation of zein. Preferential accumulation of certain globulins has been found in the mutant during the second phase of globulin formation. The increased accumulation of globulins in the opaque-2 mutant endosperm is considered a response to the arrest in zein synthesis.

Albumins↗

The evolution of polymorphic sexual systems in daffodils (Narcissus).

Narcissus, the daffodil genus, exhibits an unusual diversity of sexual systems, with populations that are monomorphic, dimorphic or trimorphic for style length. Associated with this variation are striking differences among species in floral morphology and pollination biology. This diversity provides an opportunity to investigate the evolution of mating polymorphisms, and to determine how floral morphology promotes transitions among sexual systems. Because of the absence of heteromorphic incompatibility in Narcissus, floral morphology plays a key role in governing patterns of outcrossed mating. Phylogenetic evidence indicates that stylar monomorphism is ancestral in the genus, with multiple origins of stylar polymorphism, including independent origins of stigma-height dimorphism, distyly and tristyly. Sexual polymorphisms have evolved only in lineages with narrow floral tubes that are pollinated by Lepidoptera and/or long-tongued bees. Populations of polymorphic Narcissus species are typically dominated by the long-styled morph and display imperfect reciprocity in the positions of sexual organs. These features are consequences of the unusual association between stylar polymorphism and a self-incompatibility system that permits intramorph mating.

Biological Evolution↗

Plant venereal diseases: insights from a messy metaphor.

The concept of plant venereal disease is examined from definitional, operational and axiomatic viewpoints. The transmission of many plant pathogens occurs during the flowering phase and is effected either by pollinators or by wind dispersal of spores from inflorescences. Attraction of insects by pseudo-flowers or sugary secretions also serves to spread many diseases. Given the diversity of processes involved, a simple all-encompassing parallel with animal venereal diseases is not possible. Operationally establishing the routes of disease transmission, as well as quantifying the relative magnitudes of these different routes, remains critical for understanding disease dynamics and controlling spread in agricultural contexts. From an axiomatic viewpoint, sexually transmitted diseases are characterized by frequency-dependent transmission, transmission in the adult stage, and by virulence effects involving sterility rather than mortality. These characteristics serve to differentiate the dynamics and evolution of sexually transmitted diseases from that of other diseases and are features that are also shared by many pollinator-transmitted diseases. However, the majority of plant diseases that involve the reproductive structures show a rich biology that defies easy categorization. The experimental convenience of plants and their pathogens is likely to play an important role in understanding the evolution of disease traits, irrespective of what descriptive terms are applied to the natural history of the transmission process.

Animals↗

Local genetic differentiation among populations of the mass-flowering tropical shrub Erythroxylum havanense (Erythroxylaceae).

This study is aimed to understand the role of life-history traits in determining the genetic structure of populations. We used Erythroxylum havanense, a distylous shrub with synchronous and massive patterns of flowering and fruiting. We suggest that the high concentration of ephemeral resources produced by mass flowering satiates both pollinators and frugivores, restricting gene flow and leading to genetic differentiation among populations. Using random amplified polymorphic DNAs as genetic markers, we estimated genetic diversity and structure statistics to quantify the amount and distribution of genetic variation within and among five populations from the Pacific coast of Mexico. High levels of genetic variation within populations and significant differentiation among populations located very near to each other were found. Furthermore, spatial autocorrelation analysis indicated the presence of significant genetic structure at short spatial distances. We suggest that by influencing the foraging behavior of pollinators and frugivores, mass flowering may produce the observed patterns of genetic structure, while small differences in flowering or fruiting phenology could further reinforce the isolation of nearby populations.

Animals↗

Natural selection on floral traits of female Silene dioica by a sexually transmitted disease.

Floral traits endowing high reproductive fitness can also affect the probability of plants contracting sexually transmitted diseases. We explore how variations in floral traits influence the fitness of Silene dioica females in their interactions with pollinators carrying pollen or spores of the sterilizing anther-smut fungus Microbotryum violaceum. We collected healthy and infected plants in a highly diseased population and grew them under conditions that 'cure' infected individuals, and used standard regression methods to detect natural selection on floral traits. Narrow-sense heritabilities, coefficients of additive genetic variation (CV(A)) and genetic correlations among traits were estimated from paternal half-sib groups. Pollinator preferences imposed strong direct and directional selection on traits affecting female attractiveness and pollen-/spore-capturing abilities. Levels of additive genetic variance were high in these traits, suggesting that rapid responses to selection are possible. By considering our results in the light of spatial and temporal heterogeneity resulting from the colonization dynamics typical for this species, we suggest that the conflicting selective effects of pollen/spore loads lead to the maintenance of genetic variation in these traits.

Flowers↗

Three-dimensional reciprocity of floral morphs in wild flax (Linum suffruticosum): a new twist on heterostyly.

Here, we studied the floral morphology and pollination of the distylous plant Linum suffruticosum (Linaceae) in southern Spain. We observed a previously unreported form of distyly that involved twisting and bending of styles and stamens during floral development to achieve three-dimensional reciprocity of anthers and stigmas in the long-styled (pin) and short-styled (thrum) morphs. This developmental pattern causes pin pollen to be placed on the underside of pollinating Usia flies (Bombyliidae), and thrum pollen to be placed on the top of the thorax and abdomen. The pin stigmas contact the flies on the dorsum, apparently picking up predominantly thrum pollen, and the thrum stigmas contact the flies on the ventral surface, apparently picking up predominantly pin pollen. This form of heterostyly would appear on morphological grounds to be far more efficient in dispersing pollen between compatible morphs than the typical pin-thrum system. If so, this plant fits Darwin's prediction of efficient pollen flow between heterostylous morphs more closely than anything Darwin himself reported. Molecular phylogenetic analyses indicate that this form of heterostyly evolved in a lineage that already had typical heterostyly. The analyses also indicate that there have been several independent origins of heterostyly in Linum and at least one reversal to stylar monomorphism.

Animals↗

The chemical nature of fetid floral odours in stapeliads (Apocynaceae-Asclepiadoideae-Ceropegieae).

By emitting strong fetid scents, sapromyiophilous flowers mimic brood and food sites of flies to attract them as pollinators. To date, intensive comparative scent analyses have been restricted to sapromyiophilous Araceae. Here, we analysed flower volatiles of fetid stapeliads to improve our understanding of the floral biology of fly pollinated species, and to learn whether mimicry types comparable to those found in Araceae exist. Floral volatiles of 15 species out of 11 genera within the Asclepiadoideae-Ceropegieae-Stapeliinae were collected via headspace adsorption and thermal desorption and analysed by gas chromatography-mass spectometry (GC-MS). Data were analysed using CNESS-NMDS statistics. Sapromyiophilous stapeliads are highly diverse in their scent composition, in which sulphur compounds, benzenoids, fatty acid derivatives or nitrogen-containing compounds dominate. Four groups are evident: species with high p-cresol content but low amounts of polysulphides (herbivore faeces mimicry); species with mainly polysulphides and low amounts of p-cresol (carnivore/omnivore faeces or carcass mimicry); species with high amounts of heptanal and octanal (carnivore/omnivore faeces or carcass mimicry); and species with hexanoic acid (urine mimicry). Considering the findings in the unrelated Araceae, our results support the universality of different mimicry types that are obviously subsumed under the sapromyiophilous syndrome.

Animals↗

Collinsia sparsiflora in serpentine and nonserpentine habitats: using F2 hybrids to detect the potential role of selection in ecotypic differentiation.

Here we document phenotypic differences between serpentine and nonserpentine ecotypes of Collinsia sparsiflora, as well as patterns of selection in these contrasting soil habitats. We transplanted the two parental ecotypes and experimental F2 hybrids into six field sites, and collected morphological, phenological and fitness data on emergent plants. To focus on edaphically mediated selection, rather than on pollinator-mediated selection, we used pollinator-exclusion cages. Transplanted parentals of the two ecotypes showed genetic differentiation for floral traits, but not for phenological traits or cotyledon size. For the F2 hybrids growing on serpentine soils, there was significant directional selection on cotyledon size, flower size and flower shape. However, the pattern of selection did not differ significantly between serpentine and nonserpentine sites. Overall, we did not see evidence for divergent selection on the two soil types. We conclude that differences in floral traits between the ecotypes do not reflect adaptation to physical conditions associated with soil type, and that there are unmeasured traits that must be contributing to ecotypic differentiation.

Asbestos, Serpentine↗

Investigating the independent evolution of the size of floral organs via G-matrix estimation and artificial selection.

The attractiveness of a plant to pollinators is dependent on both the number of flowers produced and the size of the petals. However, limiting resources often result in a size/number trade-off, whereby the plant can make either more flowers or larger flowers, but not both. If developmental genes underlying sepal and petal identity (some of which overlap) also influence size, then this shared genetic basis could constrain the independent evolution of floral size and attractiveness. Here, we determined whether the size of sepals and petals in the dioecious perennial, Silene latifolia, are developmentally independent by performing two experiments: a genetic variance-covariance experiment to estimate genetic correlations between calyx width, petal-limb length, flower mass, and number and a four-bout artificial-selection experiment to alter calyx width and estimate the correlated response in petal-limb length. In addition, we determined whether variation in petal-limb length is the result of cell expansion or cell proliferation. The first experiment revealed that petal-limb length is not genetically correlated with calyx width, and the second experiment confirmed this; selection on calyx width did not result in a predictable or significant change in petal-limb length. Flower number was negatively correlated with all the floral traits measured, indicating a flower size/number trade-off. Cell number, but not size, explained a significant amount of the variation in petal-limb length. We conclude that the size of the two outer floral organs can evolve independently. This species can therefore increase the number of flowers produced by decreasing investment in the calyx without simultaneously decreasing petal size and the attractiveness of each individual flower to pollinators.

Biological Evolution↗