Amplificación cruzada de microsatélites en el género Centaurea (Compositae).

11p. Fotog. en color. Tablas.[EN] Microsatellites are widely used for population genetic studies although the development of these species-specific markers is costly and time-consuming. One strategy for saving time and money is the use of markers developed for one species (source species) in a different species (target species). This is known as cross-amplification. In the present work, two sets of microsatellites are used to test their transferability to six narrow endemic Centaurea species: i) 16 nuclear loci previously published for three congeneric species and ii) 10 universal chloroplast markers designed from Nicotiana tabacum sequences. Seventeen of the 26 markers tested were transferable and 14 of them were also polymorphic and therefore useful for future works. Nuclear markers were more variable and thus more informative than chloroplast markers. Interspecific amplification performed better for the nuclear loci developed for different Centaurea species than for the universal chloroplast markers developed for Nicotiana tabacum. Likewise, transferability was more successful for the species from sect. Phalolepis than for sect. Lepteranthus. Therefore, our results support the idea that the success of the cross-amplification is influenced by the evolutionary distance between the target and the source species.[ES] Los microsatélites se usan habitualmente en estudios de genética de poblaciones aunque su desarrollo es un proceso caro y largo dada su elevada especificidad. Una estrategia que permite ahorrar tiempo y dinero es la llamada amplificación cruzada que consiste en amplificar el DNA de una especie determinada (especie objetivo) usando marcadores que han sido diseñados para una especie diferente (especie fuente). En este trabajo se ha realizado un ensayo sobre amplificación cruzada usando seis endemismos del género Centaurea y dos conjuntos de microsatélites: i) 16 marcadores nucleares desarrollados para tres especies congenéricas y ii) 10 marcadores cloroplásticos universales diseñados para Nicotiana tabacum. Diecisiete de los 26 marcadores evaluados resultaron transferibles, de los cuales 14 fueron polimórficos siendo así útiles para futuros trabajos. Las regiones nucleares se mostraron más variables y por lo tanto más informativas que las cloroplásticas. La amplificación cruzada funcionó mejor para los marcadores nucleares específicos de Centaurea que para los cloroplásticos universales específicos de Nicotiana tabacum. A su vez, se obtuvo mejor resultado para las especies de la sección Phalolepis que para las de la sección Lepteranthus. En consecuencia, nuestros resultados apoyan la idea de que el éxito de la amplificación cruzada está estrechamente ligado a la distancia evolutiva entre especie fuente y especie objetivo.Financial support from the Spanish MICINN (Project CGL2007-60781 and CGL2010/18631) and the Generalitat de Catalunya (Ajuts a Grups de Recerca Consolidats 2009/ SGR/00439-GREB) is gratefully acknowledged.Peer reviewe

Speciation and genetic diversity in Centaurea subsect. Phalolepis in Anatolia

14 p., mapas, tablasMountains of Anatolia are one of the main Mediterranean biodiversity hotspots and their richness in endemic species amounts for 30% of the flora. Two main factors may account for this high diversity: the complex orography and its role as refugia during past glaciations. We have investigated seven narrow endemics of Centaurea subsection Phalolepis from Anatolia by means of microsatellites and ecological niche modelling (ENM), in order to analyse genetic polymorphisms and getting insights into their speciation. Despite being narrow endemics, all the studied species show moderate to high SSR genetic diversity. Populations are genetically isolated, but exchange of genes probably occurred at glacial maxima (likely through the Anatolian mountain arches as suggested by the ENM). The lack of correlation between genetic clusters and (morpho) species is interpreted as a result of allopatric diversification on the basis of a shared gene pool. As suggested in a former study in Greece, post-glacial isolation in mountains would be the main driver of diversification in these plants; mountains of Anatolia would have acted as plant refugia, allowing the maintenance of high genetic diversity. Ancient gene flow between taxa that became sympatric during glaciations may also have contributed to the high levels of genetic diversity.This work was supported by the Spanish Ministerio de Ciencia e Innovación [project CGL2010/18631] and Generalitat de Catalunya [Ajuts a Grups de Recerca Consolidats 2014-SGR514-GREB]Peer reviewe

ISOLATION AND CHARACTERIZATION OF NOVEL MICROSATELLITE MARKERS FOR ARCTIUM MINUS (COMPOSITAE)

[EN] Premise of the study: Microsatellite primers were developed for the invasive plant Arctium minus to investigate the effects of facultative self-pollination and the biannual habit on population genetic structure, as well as the colonization of the Americas by this Eurasian species. Methods and Results: Sixteen di- and trinucleotide microsatellite loci were identifi ed in six populations. The number of alleles per locus ranged from one to 10, observed heterozygosities ranged from 0.000 to 0.897, and the mean value of F IS was 0.316. Conclusions: These results indicate the utility of these loci in future studies of population genetics in A. minus .Peer reviewe

Arctium L.

Género con 41 especies de Eurasia templada y norte de África, desde la Península Ibérica hasta las cordilleras del este del Asia central y desde el África norsahariana hasta el sur de Escandinavia. Algunas especies introducidas en América del Norte, Japón y Australia, y en América del Sur, una especie en la Argentina, Brasil, Chile y Uruguay.Este estudio fue realizado sobre la base del análisis de bibliografía específica, base de datos, especímenes de herbario, análisis morfométricos y viajes de campo. Como resultado se reconoció para Argentina una única especie, Arctium minus, bardana. Dicha especies es originaria de Eurasia templada y norte de África. Naturalizada en Groenlandia, Canadá, Estados Unidos de América, Japón y Australia, y en América del Sur en la Argentina, Brasil, Chile y Uruguay. Habita ambientes húmedos y sombreados, con suelos fértiles, bajos, nitrificados, comúnmente ruderales como baldíos, jardines, caminos, pasturas, márgenes de ríos, arroyos, canales y acequias, bosques perturbados y vías férreas, entre los 0 y 1000 m s. m.Peer Reviewe

Chromosome Numbers in the Genera Cousinia, Olgaea and Syreitschikovia (Compositae)

14 p. Il.[EN] The scarce karyological data available for both the Arctium-Cousinia complex and the Onopordum group has led us to provide more data essential to understand the karyological evolution of these taxa. Chromosome counts were made on somatic metaphases using the squash technique. We report 20 chromosome number records for the genus Cousinia s. lat. from the area of Near East and Central Asia. Thirteen of them are first chromosome counts for the studied species, and the remaining seven records provide confirmation of scarce or uncertain previous data. We also present the first chromosome counts for three of the 13 sections of this genus. Our records for Arctioid species show karyological uniformity with 2n=36. In the Cousinioid group, 13 records agree with the three major numbers of its characteristic dysploidy series 2n=22, 24 and 26. We report first chromosome counts for the genera Olgaea and Syreitschikovia from Kazakhstan, being 2n=26 and 24, respectively. Our results confirm a hypothesis that the Arctioid and Cousinioid clades, although forming a monophyletic group, have followed different evolutionary paths. In the Onopordum group, our results confirm the existence of two lineages; the colonizing biennial taxa are characterized by n=17, while the perennial genera have n=12, 13. The evidence for recent polyploidization is absent in both the Arctium-Cousinia complex and the Onopordum group.Peer reviewe

Genetic diversity in Silene sennenii Pau (Caryophyllaceae) assayed through DNA-based techniques

15 p.[EN]Genetic diversity in Silene sennenii Pau (Caryophylaceae) asayed through DNA-based techniques.— Silene sennenii is a narrow endemic species from the NE of the Iberian Peninsula. It is considered as EN (“Endangered”) according to the IUCN criteria and is under legal protection in Catalonia. In the present work we report an assay using three different approaches for surveying the genetic diversity in this rare, endangered campion: analysis of chloroplast haplotypes, AFLPs and transferability of microsatellites previously developed for Silene latifolia. None of the nine chloroplast regions sequenced showed any variability. Five out of the 21 microsatellite loci tested were polymorphic although more loci are required in order to perform a robust population genetics study. Regarding the AFLP analysis, five out of the 26 pairs of primers tested exhibited moderate levels of variability and therefore they could be useful for further investigating the genetic structure of S. sennenii. Although preliminary, our results based on three different genetic markers are in agreement with the low values of genetic variation already reported for this species employing allozymes.[ES]Diversidad genética de Silene sennenii Pau (Caryophyllaceae) a partir de técnicas basadas en ADN.— Silene sennenii es una especie endémica, circunscrita a un área extremadamente reducida al NE de la Península Ibérica. Está catalogada como EN («En Peligro») según criterios UICN y se encuentra legalmente protegida en Cataluña. En el presente trabajo se expone el ensayo de tres aproximaciones diferentes al estudio de su diversidad genética: análisis de haplotipos cloroplásticos, AFLPs y transferibilidad de microsatélites diseñados previamente para Silene latifolia. Ninguna de las nueve regiones cloroplásticas secuenciadas ha presentado variabilidad. Se han obtenido cinco loci microsatélites polimórficos de los 21 ensayados, cantidad insuficiente para llevar a cabo un estudio robusto sobre genética poblacional. En cuanto a AFLPs, cinco combinaciones de cebadores de las 26 probadas han mostrado niveles moderados de variabilidad siendo así útiles para posteriores trabajos sobre la estructura genética de S. sennenii. Aun siendo preliminares, los resultados aquí expuestos, fruto del uso de tres tipos de marcadores basados en ADN, confirman la escasa diversidad genética anteriormente obtenida para esta especie mediante la técnica de electroforesis de aloenzimas.This study was funded by Project CGL2007-60475/BOS from MEC (Ministerio de Educación y Ciencia, Spain) and also from grants from the Generalitat de Catalunya (Ajuts a Grups de Recerca Consolidats 2009/SGR/00439-GREB). In addition, J. L.-P. was supported by a post-doctoral “Beatriu de Pinós” fellowship (Generalitat de Catalunya) and S. M. A. was supported by a pre-doctoral ADR fellowship (Universitat de Barcelona).Peer reviewe

Systematics of the Arctioid group: Disentangling Arctium and Cousinia (Cardueae, Carduinae)

16p. Incluye suplemento que contiene: Appendix 2. Species examined. Taxon, origin and voucher information. [http://www.ingentaconnect.com/content/iapt/tax/2011/00000060/00000002/art00020/supp-data/11677lopezsuppl][EN] We investigated the phylogeny of the Arctioid lineage of the Arctium-Cousinia complex in an attempt to clarify the conflictive generic boundaries of Arctium and Cousinia. The study was based on analyses of one nuclear (ITS) and two chloroplastic (trnL-trnT-rps4, rpl32-trnL) DNA regions of 37 species and was complemented with morphological evidence where possible. Based on the results, a broadly redefined monophyletic genus Arctium is proposed. The subgenera Hypacanthodes and Cynaroides are not monophyletic and are suppressed. In contrast, the traditional sectional classification of the genus Cousinia is maintained. The genera Anura, Hypacanthium and Schmalhausenia are reduced to sectional level.Peer reviewe

11677_Lopez.indd

Abstract We investigated the phylogeny of the Arctioid lineage of the Arctium-Cousinia complex in an attempt to clarify the conflictive generic boundaries of Arctium and Cousinia. The study was based on analyses of one nuclear (ITS) and two chloroplastic (trnL-trnT-rps4, rpl32-trnL) DNA regions of 37 species and was complemented with morphological evidence where possible. Based on the results, a broadly redefined monophyletic genus Arctium is proposed. The subgenera Hypacanthodes and Cynaroides are not monophyletic and are suppressed. In contrast, the traditional sectional classification of the genus Cousinia is maintained. The genera Anura, Hypacanthium and Schmalhausenia are reduced to sectional level

Phylogeny and evolution of the Arctium-Cousinia complex (Compositae, Cardueae-Carduinae)

[EN] The phylogeny and evolution of the Arctium-Cousinia complex, including Arctium, Cousinia as one of the largest genera of Asteraceae, Hypacanthium and Schmalhausenia, is investigated. This group of genera has its highest diversity in the Irano-Turanian region and the mountains of Central Asia. We generated ITS and rpS4-trnT-trnL sequences for altogether 138 species, including 129 (of ca. 600) species of Cousinia. As found in previous analyses, Cousinia is not monophyletic. Instead, Cousinia subgg. Cynaroides and Hypacanthodes with together ca. 30 species are more closely related to Arctium, Hypacanthium and Schmalhausenia (Arctioid clade) than to subg. Cousinia (Cousinioid clade). The Arctioid and Cousiniod clades are also supported by pollen morphology and chromosome number as reported earlier. In the Arctioid clade, the distribution of morphological characters important for generic delimitation, mainly leaf shape and armature and morphology of involucral bracts, are highly incongruent with phylogenetic relationships as implied by the molecular data. No taxonomic solution for this conflict can be offered, and the characters named are interpreted as homoplasious. Although phylogenetic resolution in the Cousinioid clade is poor, our ITS and rpS4-trnT-trnL sequences contain some phylogenetic information. For example, the six annual species of the Cousinioid clade fall into two groups. Poor phylogenetic resolution probably results from lack of characters and the high number of taxa in this species-rich and comparatively young (ca. 8.7 mya) lineage. We hypothesize that speciation in the Cousinioid clade was mainly allopatric.Peer reviewe

The Polyploid Series of the Achillea millefolium Aggregate in the Iberian Peninsula Investigated Using Microsatellites

10 p., gráf., fot., mapasThe Achillea millefolium aggregate is one of the most diverse polyploid complexes of the Northern hemisphere and has its western Eurasian boundary in the Iberian Peninsula. Four ploidy levels have been detected in A. millefolium, three of which have already been found in Iberia (diploid, hexaploid and octoploid), and a fourth (tetraploid) reported during the preparation of this paper. We collected a sample from 26 Iberian populations comprising all ploidy levels, and we used microsatellite markers analyzed as dominant in view of the high ploidy levels. Our goals were to quantify the genetic diversity of A. millefolium in the Iberian Peninsula, to elucidate its genetic structure, to investigate the differences in ploidy levels, and to analyse the dispersal of the species. The lack of spatial genetic structure recovered is linked to both high levels of gene flow between populations and to the fact that most genetic variability occurs within populations. This in turn suggests the existence of a huge panmictic yarrow population in the Iberian Peninsula. This is consistent with the assumption that recent colonization and rapid expansion occurred throughout this area. Likewise, the low levels of genetic variability recovered suggest that bottlenecks and/or founder events may have been involved in this process, and clonal reproduction may have played an important role in maintaining this genetic impoverishment. Indeed, the ecological and phenologic uniformity present in the A. millefolium agg. in Iberia compared to Eurasia and North America may be responsible for the low number of representatives of this complex of species present in the Iberian Peninsula. The low levels of genetic differentiation between ploidy levels recovered in our work suggest the absence of barriers between them.Dr Joan Uriach Marsal privately funded this research (grant assigned to AS and NGJ). Generalitat de Catalunya partly contributed financial support by grants "Ajuts a Grups de Recerca Consolidats" number 2009SGR/00439 and 2014/SGR/514 to all the authors.Peer reviewe