Protein relationships between natural populations ofUlex europaeus andU. gallii (Faboideae,Genisteae) and their hybrids

Prostrate ecotypes ofUlex gallii, an atlantic species with autumnal flowering, grow on capes in Brittany (France).U. europaeus, a widespread species, has winter and spring flowering, but some plants can precociously flower in autumn, so that hybridization can be possible between these two species in areas where their ranges overlap. By using different isoenzyme systems and crossed immunoelectrophoresis obtained with seed proteins, it is possible to consider plants with intermediate morphological characters betweenU. gallii andU. europaeus as natural hybrids.     

Plant phenology and seed predation: interactions between gorses and weevils in Brittany (France)

Exapion ulicis (Forster) and Exapion lemovicinum (Hoffmann) (Coleoptera: Curculionoidea: Apionidae) are seed predators of the three gorse species occurring in Brittany (France): Ulex europaeus L., Ulex gallii Planch., and Ulex minor Roth.(Fabaceae). Host‐plant phenology plays a major role in the relationship between apionid weevils and their gorse species, because larvae develop within gorse pods and adults have to wait for pod dehiscence to be released. We monitored flowering and fruiting phenology of gorse species, weevil reproductive behaviour, and egg‐laying patterns in six natural populations in the native area of these gorse species. At each site, U. europaeus, which flowers mainly in spring, was sympatric with one of two autumn flowering gorse species, U. gallii and U. minor. We noticed that E. ulicis laid eggs in spring and was restricted to U. europaeus whereas E. lemovicinum laid eggs in autumn and was restricted to the two autumn‐flowering species U. gallii and U. minor. Therefore, host specificity depended on gorse phenology, and not on geographic proximity. In addition, the infested pod content showed that E. ulicis laid several eggs per pod and suggested that females chose pods with the highest numbers of seeds. In contrast, E. lemovicinum laid a single egg per pod and showed no preference for pods with many seeds. Finally, the impact of seed predation by E. ulicis was higher than that of E. lemovicinum.     

Chromosome numbers and geographic distribution of Ulex gallii and U. minor (Leguminosae)

FERNANDEZ PRIETO, J. A., NAVA, H. S., VERA, M. L., ALVAREZ MARTINEZ, M. J., DIAZ, T. E., FERNANDEZ CASADO, M. A., FERNANDEZ-CARVAJAL, M. C. & GUTIERREZ VILLARIAS, M. I., 1993. Chromosome numbers and geographic distribution of Ulex gallii and U. minor (Leguminosae). Chromosome numbers from 188 samples of the Ulex gr. gallii-minor from Portugal, Spain, France and Great Britain are reported. The gametic and somatic chromosome numbers are n = 16 and In = 32, respectively, in all samples of U. minor. Three levels of ploidy (diploid, tetraploid and hexaploid) have been detected in samples of U. galli, sensu lata from N Spain. In samples from Brittany and Great Britain, only the tetraploid level has been found. New data are compared with all previous reports for the species of this group of gorses.     

Hordein polymorphism in diploid and tetraploid Mediterranean populations of the Hordeum murinum L. complex

Polymorphism analyses of the hordeins, main storage proteins in barley, were conducted on 35 natural populations of Hordeum murinum s.l. from North Africa; this specific complex includes three subspecies with two ploidy levels: H. murinum subsp. glaucum (2n=2x=14), H. murinum subsp. leporinum and subsp. murinum (2n=4x=28). Twenty of these populations belong to the diploid subsp. glaucum, 14 other tetraploid populations belong to the subsp. leporinum. In addition, six populations of the tetraploid murinum were sampled in France: two along the Mediterranean coast and four in Brittany. The polymorphism observed in the electrophoretic patterns highlights strong correlations between bioclimatic features and di- and tetraploid taxa distribution. Moreover, the variation was not randomly distributed within the different ploidy levels, and is correlated with environmental factors. The ecological differentiation of the two main taxa, H. murinum subsp. leporinum and subsp. glaucum is clearly highlighted.     

Crossing relationships and chromosome numbers of Solanum section Solanum in Uganda

The ability of taxa to cross/hybridize is useful information for plant systematists and breeders. Crossability reflects reproductive isolation and the biological species concept stresses the need for reproductive isolation between species to maintain morphological distincness. For plant breeders knowledge on crossing ability facilitate selection of taxa for character improvement breeding. In this study, the crossing relationships and chromosome numbers within and among Ugandan species of Solanum sect. Solanum is studied by making 800 crosses involving 246 combinations. Less than half of these combinations were successful, producing F1 offspring. All studied accessions are self‐compatible and most accessions crossed readily with accessions of their own species. Interspecific crossings failed either to yield seeds, yielded F1 seeds that did not germinate, or resulted in F1s that did not have stainable pollen – implying a crossing barrier; or stainable pollen, but with chromosome numbers that indicated reproduction by apomixis. The results support the taxonomic treatment of Solanum based on classical, numerical and partly molecular evidences. The material studied represents eight Ugandan taxa: S. americanum, a diploid (2n = 2x = 24); five tetraploids (2n = 4x = 48) S. florulentum, S. memphiticum, S. tarderemotum, S. villosum ssp. villosum and S. villosum ssp. miniatum; and two hexaploids (2n = 6x = 72) S. scabrum subsp. scabrum and S. scabrum subsp. laevis. In addition to confirming the ploidy levels of the Ugandan accessions, the ploidy levels of S. florulentum, S. memphiticum and S. tarderemotum are reported for the first time. Non‐Ugandan material of Solanum sarrachoides was found to be diploid. Knowledge of the crossing behaviour and ploidy levels in Solanum will facilitate breeding for character improvement in these important species that are used commonly as food and/or medicine in eastern Africa.     

The association among herbivory tolerance,ploidy level,and herbivory pressure in <Emphasis Type="Italic">cardamine pratensis</Emphasis>

We tested whether differences in ploidy level and previous exposure to herbivory can affect plant tolerance to herbivory. We conducted a common garden experiment with 12 populations of two ploidy levels of the perennial herb Cardamine pratensis (five populations of tetraploid ssp. pratensis and seven populations of octoploid ssp. paludosa). Earlier studies have shown that attack rates by the main herbivore, the orange tip butterfly Anthocharis cardamines, are lower in populations of octoploids than in populations of tetraploids, and vary among populations. In the common garden experiment, a combination of natural and artificial damage significantly reduced seed and flower production. We measured tolerance based on four plant-performance metrics: survival, growth, seed production and clonal reproduction. For three of these measurements, tolerance of damage did not differ between ploidy levels. For clonal reproduction, the octoploids had a higher tolerance than the tetraploids, although they experience lower herbivore attack rates in natural populations. Populations from sites with high levels of herbivory had higher tolerance, measured by seed production, than populations with low levels of herbivory. We did not detect any significant costs of tolerance. We conclude that high intensity of herbivory has selected for high tolerance measured by seed production in C. pratensis.     

Population responses of Ulex shrubs to fire in a lowland heath community

Question: How does the frequency of heathland fire events affect population growth rates of two woody shrub species, Ulex gallii and U. minor? Location: Dry heathland on the south coast of England, UK. Methods: The population dynamics of U. gallii and U. minor were modelled at each phase of the heathland cycle — pioneer, building, mature and degenerate — using periodic matrix products to investigate the response to different fire regimes. Results: Population growth rates of both Ulex species declined under annual burning. Initially, as the time between burns increased, population growth rate increased for both species. Maximum population growth rates for each Ulex species were achieved under a 16‐yr fire return interval. Fire return intervals > 16 yr resulted in declining population growth rates. Conclusions: A species‐specific critical fire frequency can be predicted, the minimum fire return intervals permitting persistence were 4 yr for U. minor and 3 yr for U. gallii. These patterns are similar to those reported for a range of woody plant species within savanna environments.     

Butterfly seed predation: effects of landscape characteristics,plant ploidy level and population structure

Polyploidization has been suggested as one of the most common mechanisms for plant diversification. It is often associated with changes in several morphological, phenological and ecological plant traits, and therefore has the potential to alter insect–plant interactions. Nevertheless, studies evaluating the effect of plant polyploidy on interspecific interactions are still few. We investigated pre-dispersal seed predation by the butterfly Anthocharis cardamines in 195 populations of two ploidy levels of the herb Cardamine pratensis (tetraploid ssp. pratensis, 2n = 30 vs. octoploid ssp. paludosa, 2n = 56–64). We asked if differences in incidence and intensity of predation among populations were related to landscape characteristics, plant ploidy level and population structure. The incidence of the seed predator increased with increasing plant population size and decreasing distance to nearest population occupied by A. cardamines. The intensity of predation decreased with increasing plant population size and was not affected by isolation. Probability of attack decreased with increasing shading, and intensity of predation was higher in grazed than in non-grazed habitats. The attack intensity increased with increasing mean flower number of plant population, but was not affected by flowering phenology. Individuals in tetraploid populations suffered on average from higher levels of seed predation, had higher mean flower number, were less shaded and occurred more often in grazed habitats than octoploid populations. When accounting for differences in habitat preferences between ploidy levels there was no longer a difference in intensity of predation, suggesting that the observed differences in attack rates among populations of the two ploidy levels are mediated by the habitat. Overall, our results suggest that polyploidization is associated with differentiation in habitat preferences and phenotypic traits leading to differences in interspecific interaction among plant populations. This, in turn, may facilitate further divergence of ploidy levels.     

Flow cytometric evidence for multiple ploidy levels in the endosperm of some gymnosperm species

 Considered to be haploid tissue, the endosperm of coniferous trees has been extensively used by forest geneticists. Using laser flow cytometry, we show that endosperm ploidy level depends on the systematic position. The Abies, Cedrus and Pinus species tested exhibited uniform haploid endosperm compared to the diploid DNA content of the corresponding embryo. Endosperm of Cupressaceae contained multiple ploidy levels: Cupressus arizonica, Juniperus oxycedrus and Thuja orientalis endosperms exhibited a mixture of haploid–diploid nuclei, while C. atlantica and C. sempervirens endosperms contained six ploidy levels: 1C, 2C, 3C, 4C, 5C and 6C. Physiological and genetic implications of this original feature are discussed. Received: 17 August 1996 / Accepted: 18 October 1996     

Phylogenetic analysis of Hordeum marinum Huds. based on nucleotide sequences linked to the vrs1 locus

 More than 100 accessions of Hordeum marinum ssp. marinum, ssp. gussoneanum 2x and ssp. gussoneanum 4x were studied by analyzing the nuclear DNA homologous to the chloroplast translation elongation factor. PCR amplification using the locus-specific primers generated a 932 bp fragment from ssp. marinum, a 915 bp fragment from ssp. gussoneanum 2x, and 915 bp and 931 bp fragments from ssp. gussoneanum 4x. Sequencing of the entire DNAs showed that the 915-bp fragments of diploids and tetraploids are identical, indicating that the diploid probably is the immediate donor of the 915-bp fragment of tetraploids. Although the donor of the 931-bp fragment was not determined, phylogenetic analysis of the data showed that the clade of the 931-bp fragment was included in the H. marinum complex. A MspI site was diagnostic to distinguish ssp. marinum into two clades with disjunct distributions, the Iberian Peninsula (type 2) and other regions (type 1). Nucleotide variation was revealed in the type 2 accessions but all the other clades were highly uniform. Received January 3, 1999 Accepted January 24, 2001     

Correlation between nuclear DNA values and differing optimal ploidy levels inNarcissus,Hyacinthus andTulipa cultivars

4C nuclear DNA amounts were determined in 16 large decorative cultivars ofNarcissus (Amaryllidaceae), 13 ofHyacinthus (Hyacinthaceae) and 12 ofTulipa (Liliaceae) at different levels of ploidy. Within each genus, nuclear DNA amounts and ploidy levels are positively correlated, with no DNA loss in polyploids.Based on wide surveys of chromosome numbers, the maximum numbers of cultivars, interpreted as the optimum levels of selective success or horticultural fitness, were found to be at the tetraploid level inNarcissus (2n=4x=28), the triploid inHyacinthus (2n=3x=24) and the diploid inTulipa (2n=2x=24). All these ploidy optima were shown to correspond to a small range of nuclear DNA amounts (4C=96-139 pg), which could suggest the existence of a single DNA value optimal for the three biologically similar but unrelated genera. In each case the optimum is at an equilibrium reached between enhanced size and other morphological characteristics on one hand and reduced growth rate on the other, both resulting from increase in ploidy and nuclear DNA amounts.     

Relationship between ploidy level and genome size in strawberries

This study investigated the pattern of variation in nuclear DNA content at different ploidy levels in Fragaria (Strawberry, Rosaceae) using flow cytometry based on mean fluorescent intensity (MFI) reflected by propidium-iodide-stained nuclei. On average, MFI values were 237 for diploids F. vesca, F. viridis, and F. nubicola, 416.5 for tetraploid F. orientalis, 621.5 for hexaploid F. moschata, and 798 for octoploids F. × ananassa, F. virginiana, and F. chiloensis. Within diploids MFI ranged from 225.9 in F. vesca ssp. vesca to 255.4 in F. nubicola, and within octoploids varied from 766 in F. × ananassa to 808 in F. virginiana. The nuclear DNA variation was significant among diploid species (N = 21, P < 0.008), but not across octoploid species (N = 17, P>0.386). MFI values were also variable among different genotypes of a given species though not significant. The values of mean basic genome DNA (MFI divided by ploidy level) were 118.5, 104, 103.5, and 99.8, respectively, for diploids, tetraploid, hexaploid, and octoploid species. This indicates that relative genomic size decreases by increasing ploidy level, and that there is no direct proportional relationship between DNA content and ploidy levels in Fragaria, supporting the idea of genome downsizing during polyploidization in plants.     

How is the invasive gorse <Emphasis Type="Italic">Ulex europaeus</Emphasis> pollinated during winter? A lesson from its native range

Many examples of plant-insect interactions have shown that selection from herbivores can act on flowering and fruiting phenology. In Ulex europaeus (Fabaceae), escaping seed predation resulted in extended, but variable flowering periods, with some plants flowering from autumn until spring and others flowering only in spring. The present study aims at understanding how gorses can have a high reproductive success during winter despite harsh climatic conditions and low number of pollinators. We measured pollen production, flower size and seed production in spring and winter, and compared the different seasons. The pollination success of flowers was high in both seasons. The flowers produced as much pollen, and were of comparable size in spring and winter, but they stayed open twice as long in winter than in spring. The high pollination rate we observed was thus due to the longer opening period of flowers and the high attractiveness of flowers during winter. However, pod abortion was higher in winter, with 43% of the flowers in winter and 75% in spring producing ripe pods. Antagonistic selective pressures exerted by biotic and abiotic interactions may, therefore, have lead to the observed flowering polymorphism, and allow U. europaeus to thrive in various climates, thus, increasing its invasiveness in different countries.     

Visual selection and maintenance of the cell lines with high plant regeneration ability and low ploidy level in Dianthus acicularis by monitoring with flow cytometry analysis

Efficient plant regeneration system from cell suspension cultures was established in D. acicularis (2n=90) by monitoring ploidy level and visual selection of the cultures. The ploidy level of the cell cultures closely related to the shoot regeneration ability. The cell lines comprising original ploidy levels (2C+4C cells corresponding to DNA contents of G1 and G2 cells of diploid plant, respectively) showed high regeneration ability, whereas those containing the cells with 8C or higher DNA C-values showed low or no regeneration ability. The highly regenerable cell lines thus selected consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.     

The application of the n-alkane technique for estimating the composition of diets consumed by equines and cattle feeding on upland vegetation communities

The application of n-alkanes as faecal markers to estimate diet composition of equines and cattle was studied. Twelve mature crossbreed mares (385 ± 47 kg live weight – LW) and six adult non-lactating cows (499 ± 36 kg LW) of Asturiana de los Valles breed were divided in groups of three animals (H1, H2, H3 and H4 in equines and C1 and C2 in cattle) and housed in individual stalls. Animals received a daily total amount of 1.0 kg DM/100 kg LW of different experimental diets composed of herbaceous (Lolium perenne L.) and woody species (Ulex gallii Planchon and heather: Erica spp. and Calluna vulgaris L.). Diet composition varied among treatments: H1 and C1 – L. perenne as sole component; H2 and C2 – L. perenne (0.70) and heather (0.30); H3 – L. perenne (0.70) and U. gallii (0.30); H4 – L. perenne (0.40), heather (0.30) and U. gallii (0.30).     

Karyotype analysis and DNA content in some species of Chrysolaena (Vernonieae,Asteraceae)

Cytogenetic characterization by karyotyping and determination of DNA content by flow cytometry of five species of Chrysolaena (Vernonieae, Asteraceae) was performed. This is the first study of nuclear DNA content realized in the genus. The 2C-values were compared with the ploidy level and the total karyotype length (TKL) of each species. Mitotic analysis revealed a base chromosome number x = 10 for all entities and different ploidy levels, from diploid (2n = 2x = 20) to octoploid (2n = 8x = 80). All species showed bimodal karyotypes composed of metacentric and submetacentric chromosomes. The average chromosome size (ML) varied from 1.86 μm to 2.70 μm, while the TKL ranged from 18.65 μm to 80.55 μm. The intrachromosomal asymmetry index (A₁) varied from 0.27 to 0.38, while the interchromosomal asymmetry index (A₂) ranged from 0.19 to 0.25. A new cytotype is reported for the first time for C. propinqua. Accessory chromosomes found in C. verbascifolia, C. cognata, C. flexuosa, and C. propinqua are also reported as new.     

Flow cytometric evidence for endopolyploidy in seedlings of some Brassica species

Flow cytometric analysis of the nuclear DNA contents of somatic tissues from seedlings of Brassica rapa L. and B. oleracea L. revealed extensive endoreduplication, resulting in tissues that contain cells with multiple ploidy levels (also called ’endopolyploidy’). Multiples of the haploid nuclear genome complement (1C) corresponding to 2C, 4C, 8C, 16C, 32C and 64C were observed in Brassica rapa, while B. oleracea exhibited a mixture of cells with five ploidy levels, 2C, 4C, 8C, 16C and 32C. The distribution of cells with the different ploidy levels was tissue-specific and characteristic of the stage of development. Multiploidy was not found in the embryos of dry seeds. Rapid endoreduplication occurred during seedling development. It is most probable that multiploidy is, if not a general feature, at least very common in Brassica species. The physiological and genetic implications of this original feature are discussed. Received: 17 March 2000 / Accepted: 17 April 2000     

Characterization of polymorphic microsatellite loci in the tropical ant‐plant Leonardoxa africana (Fabaceae: Caesalpinioideae)

Ten microsatellite loci were isolated from the African ant‐plant Leonardoxa africana (Fabaceae: Caesalpinioideae). They differentiate the two most divergent subspecies of this polytypic complex, L. a. africana and L. a. gracilicaulis, showing promise for the study of gene flow. In each of these two subspecies, high levels of within‐population variation were observed, with a number of alleles ranging from one to 10 in ssp. africana and from one to 7 in ssp. gracilicaulis, and heterozygosity from 0 to 0.933 in ssp. africana and to 0.867 in ssp. gracilicaulis. All loci amplified successfully in the two other subspecies of L. africana.     

Dental Ontogeny in Macroscelides proboscideus (Afrotheria) and Erinaceus europaeus (Lipotyphla)

We investigated the state of dental eruption in specimens of Macroscelides proboscideus and Erinaceus europaeus of known age. When M. proboscideus reaches adult size and sexual maturity, few or none of its replaced permanent cheek teeth have erupted. The approximate sequence of upper tooth eruption is P1, [I3, C, M1], [I1–2], M2, P4, [P2, P3]. Chronologically, E. europaeus erupts its molars and most premolars prior to M. proboscideus; but its first two upper incisors erupt after those of M. proboscideus, and its canines erupt around the same time. The approximate sequence of upper tooth eruption in E. europaeus is [M1, M2, P2, I3], C, M3, P4, P3, I2, I1. Unlike M. proboscideus, E. europaeus does not reach adult size until all permanent teeth except for the anterior incisors have erupted. While not unique among mammals, the attainment of adult body size prior to complete eruption of the permanent cheek teeth is particularly common among macroscelidids and other afrotherians.