Dissimilar molecular and morphological patterns in an introgressed peripheral population of a sand dune species (Armeria pungens, Plumbaginaceae).

Introgression is a poorly understood evolutionary outcome of hybridisation because it may remain largely undetected whenever it involves the transfer of small parts of the genome from one species to another. Aiming to understand the early stages of this process, a putative case from the southernmost border of the Armeria pungens range from its congener A. macrophylla is revisited following the discovery of a subpopulation that does not show phenotypic signs of introgression and resembles typical A. pungens. We analysed morphometrics, nuclear ribosomal DNA ITS and plastid DNA (trnL-trnF) sequences, genome size, 45S and 5S rDNA loci-FISH data and nrDNA IGS sequences. Within the study site, most individuals match morphologies of either of the two hybridising species, particularly the new subpopulation, with intermediate phenotypes being scarce. This pattern does not fully fit molecular evidence revealing two ITS ribotypes co-occurring intragenomically in most plants from the study site and one single plastid haplotype. Genome size and structural features of the IGS sequences both indicate that A. pungens from the study site is genetically more similar to its sympatric congener than to the remainder of its conspecifics. Introgression of A. macrophylla into A. pungens and plastid capture explain all the evidence analysed. However, important features to understand the origin and fate of the introgressed population, such as the degree and direction of introgression, which are important for understanding early stages of hybridisation in plants with low reproductive barriers, should be addressed with new data.

Past climate changes facilitated homoploid speciation in three mountain spiny fescues (Festuca, Poaceae).

Apart from the overwhelming cases of allopolyploidization, the impact of speciation through homoploid hybridization is becoming more relevant than previously thought. Much less is known, however, about the impact of climate changes as a driven factor of speciation. To investigate these issues, we selected Festuca picoeuropeana, an hypothetical natural hybrid between the diploid species F. eskia and F. gautieri that occurs in two different mountain ranges (Cantabrian Mountains and Pyrenees) separated by more than 400 km. To unravel the outcomes of this mode of speciation and the impact of climate during speciation we used a multidisciplinary approach combining genome size and chromosome counts, data from an extensive nuclear genotypic analysis, plastid sequences and ecological niche models (ENM). Our results show that the same homoploid hybrid was originated independently in the two mountain ranges, being currently isolated from both parents and producing viable seeds. Parental species had the opportunity to contact as early as 21000 years ago although niche divergence occurs nowadays as result of a climate-driven shift. A high degree of niche divergence was observed between the hybrid and its parents and no recent introgression or backcrossed hybrids were detected, supporting the current presence of reproductive isolation barriers between these species.

Balearic insular isolation and large continental spread framed the phylogeography of the western Mediterranean Cheirolophus intybaceus s.l. (Asteraceae).

Recent Quaternary geological and climate events have shaped the evolutionary histories of plant species in the Mediterranean basin, one of the most important hotspots of biodiversity. Genetic analyses of the western Mediterranean Cheirolophus intybaceus s.l. (Asteraceae) based on AFLP were conducted to establish the relationships between its close species and populations, to reconstruct the phylogeography of the group and to analyse potential unidirectional versus bidirectional dispersals between the Ibero-Provençal belt and the Balearic Islands. AFLP data revealed two main genetic groups, one constituted by the Balearic populations and Garraf (NE Iberia) and the other formed by the remaining mainland populations that were further sub-structured into two geographically separated subgroups (SE + E Iberia and NE Iberia + SW France). Genetic diversity and spatial structure analyses suggested a mid-Pleistocene scenario for the origin of C. intybaceus in southern Iberia, followed by dispersal to the north and a single colonisation event of the Balearic archipelago from the near Dianic NE Iberian area. This hypothesis was supported by paleogeographic data, which showed the existence of terrestrial connections between the continent and the islands during the Middle-Late Pleistocene marine regressions, whereas the more recent single back-colonisation of the mainland from Mallorca might be explained by several hypotheses, such as long-distance dispersal mediated by migratory marine birds or sea currents.

Swarm of terminal 35S in Cheirolophus (Asteraceae, Centaureinae).

Island radiation constitutes a playground for species diversification, which has long fascinated researchers and still does today. Because only a small subset of taxa within the pool of island colonizers is concerned by this process, the question is raised on whether some factors could make a taxon prone to radiate. Cheirolophus is the only genus of Centaureinae subtribe to have experienced a radiation in the Canary Islands. Cytogenetic characterization through FISH of 5S and 35S ribosomal RNA genes in eight Cheirolophus species from continent and Canary Islands revealed an unusually high number of 35S predominantly at terminal position, together with a single interstitial 5S rDNA locus in all the studied taxa. Such an abundance of 35S rDNA signals is unique among Centaureinae and predates Cheirolophus arrival in Canary Islands. The possible link of the rDNA profile with radiation process is discussed through a comparison with two other case studies, the closely related Rhaponticum group and the genus Centaurea.

Genome size dynamics in Artemisia L. (Asteraceae): following the track of polyploidy.

Polyploidy is a key factor in the evolution of higher plants and plays an important role in the variation of plant genomes, leading to speciation in some cases. During polyploidisation, different balancing processes take place at the genomic level that can promote variation in nuclear DNA content. We estimated genome size using flow cytometry in 84 populations of 67 Artemisia species and one population of Crossostephium chinense. A total of 73 sequences of nrDNA ITS and 3'-ETS were newly generated and analysed, together with previously published sequences, to address the evolution of genome size in a phylogenetic framework. Differences in 2C values were detected among some lineages, as well as an increase of genome size heterogeneity in subgenera whose phylogenetic relationships are still unclear. We confirmed that the increase in 2C values in Artemisia polyploids was not proportional to ploidy level, but 1Cx genome size tended to decrease significantly when high ploidy levels were reached. The results lead us to hypothesise that genome size in polyploids tends to a maximum as it follows saturation behaviour, in agreement with the Michaelis-Menten model. We tested different arithmetic functions with our dataset that corroborated a non-linear relationship of genome size increase in polyploids, allowing us to suggest a theoretical upper limit for the DNA content of this genus.

Molecular cytogenetics of Xeranthemum L. and related genera (Asteraceae, Cardueae).

Seven representatives of the genera Amphoricarpus, Chardinia, Siebera, and Xeranthemum, all of them closely related as demonstraded by molecular phylogeny, have been studied from a cytogenetic perspective. Morphometrical karyotype parameters were calculated and idiograms obtained. Fluorochrome banding was performed with chromomycin A (3) to identify GC-rich regions in the chromosomes. Fluorescence in situ hybridization allowed us to locate the sites of 18S-5.8S-26S and 5S rDNA. Silver nitrate staining was used to count the number of nucleoli and to detect the active nucleolar organizing regions. Systematic and evolutionary issues are addressed in the light of these data.

Multiple introductions from the Iberian peninsula are responsible for invasion of Crupina vulgaris in western North America.

• Crupina vulgaris is a relatively recent invader to grasslands and other open habitats in western North America. Like related Centaurea species, it was introduced from the Mediterranean region, where it does not exhibit ruderal behavior. Determining the number and sources of invasion founders allows fuller interpretation of colonization dynamics and recognition of potential intercontinental carriers, both critical factors for curbing the spread of invasive species. • We chose the molecular technique of random amplified polymorphic DNA (RAPD) to identify the number and sources of invasion founders from the eastern hemisphere, by comparing indigenous and invasive populations. • Our results indicated that the five North American populations derived from three or more successful invasion events whose founders originated in the Iberian peninsula. • Also inferred by the similarity clustering among eastern hemisphere populations is a more ancient origin of the genus to the east of the Mediterranean, a concept supported by the scarcity of suitable nonanthropic habitat in Spain. Its epizoochoric association with migratory movements of domestic herds suggests probable routes of migration first to southern Europe, then later to North America.