Núria Garcia-Jacas

The cardueae (Compositae) revisited : Insights from its, trnL-trnF, and matK nuclear and chloroplast DNA analysis

Abstract The new outline of relationships in basal branches of the family Compositae Giseke confirms that the sister group to the tribe Cardueae Cass. are not Mutisieae Cass., but rather a group of African genera now classified as the tribe Tarchonantheae Kostel. This change implies that the monophyly of the Cardueae must be reassessed on a molecular basis. Moreover, new collections in recent years allow us to extend our sampling to 70 of the 74 genera of the tribe. We performed a new molecular study of the tribe using one nuclear region (ITS) and two chloroplastic markers (trnL-trnF and matK) in addition to a more appropriate outgroup. Our results confirm that the Cardueae is a natural group but indicate some changes in subtribal delineation: the subtribe Cardopatiinae Less. is recognized and some genera are moved to other subtribes (Myopordon Boiss., Nikitinia Iljin, Syreitschikovia Pavlov, and the Xeranthemum L. group). A recapitulation of a number of interesting questions that remain unresolved in the classification of some large genera is presented.

The natural delimitation ofCentaurea (Asteraceae: Cardueae): ITS sequence analysis of theCentaurea jacea group

Taxonomic complexity has hindered partitioning the genusCentaurea into natural subdivisions, even though it has long been recognized as an unwieldy, artificial assemblage. Most of the remaining difficulties center in theCentaurea jacea group, whose taxa share a common advanced type of pollen. Because it comprises half the species of the genusCentaurea, as well as five other disputed genera previously segregated fromCentaurea (Chartolepis, Cheirolepis, Cnicus, Grossheimia andTomanthea), theCentaurea jacea group is a significant taxonomic challenge. Newer molecular approaches are useful for resolving complex relationships because they provide more precise inferences of evolutionary relationships than traditional morphological characters. Sequences of the Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA were analyzed for a comprehensive sample of this group. Results indicated that theCentaurea jacea group is monophyletic and includes the segregated genera, but not two other genera (Oligochaeta andZoegea), whose inclusion in theCentaurea jacea group was doubtful. In addition to pollen morphology, the ITS phylogeny is also supported by karyological evidence and by good correlation with biogeographic distribution of the species. The monophyly of theCentaurea jacea group suggests that a natural delimitation ofCentaurea that minimizes nomenclatural changes is possible, but only if a new type of the genus is designated.

Generic delimitation and phylogeny of the Carduncellus-Carthamus complex (Asteraceae) based on ITS sequences

Within the Mediterranean complexCarduncellus-Carthamus, taxonomic classification has proven problematic. Numerous attempts to clarify the relative systematic boundaries have included splittingCarduncellus andCarthamus into several genera, but none of these proposed classifications have been generally accepted. For a comprehensive resolution of the relationships within this group, we used sequences of the Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA. The results indicate that the complex should be classified into four genera:Carduncellus, Carthamus, Femeniasia andPhonus. The relationship between the western group (Carduncellus, Femeniasia andPhonus) and the eastern genusCarthamus are not resolved by analysis of ITS sequences, but the two groups are probably not close relatives. The ITS classifications corresponded with biogeography and less with morphological characters, which have also been the main source of confusion in traditional classifications. Most of the unusual morphological features in theCarduncellus-Carthamus complex appear to be reversals to ancestral character states.

Reconstructing the history of Campanulaceae with a Bayesian approach to molecular dating and dispersal–vicariance analyses

We reconstruct here the spatial and temporal evolution of the Campanula alliance in order to better understand its evolutionary history. To increase phylogenetic resolution among major groups (Wahlenbergieae-Campanuleae), new sequences from the rbcL region were added to the trnL-F dataset obtained in a previous study. These phylogenies were used to infer ancestral areas and divergence times in Campanula and related genera using a Bayesian approach to molecular dating and dispersal-vicariance analyses that takes into account phylogenetic uncertainty. The new phylogenetic analysis confirms Platycodoneae as the sister group of Wahlenbergieae-Campanuleae, the two last ones inter-graded into a well-supported clade. Biogeographic and dating analyses suggest that Western Asia and the Eastern Mediterranean have played a major role as centers of migration and diversification within the Campanula alliance, probably in relation to the intense orogenic activity that took place in this region during the Late Neogene, and that could have promoted isolation and allopatric speciation within lineages. Diversification rates within several Campanula lineages would have increased at the end of the Miocene, coinciding with the Messinian Stage. Strong selective pressures from climate changes and the expansion of mountainous regions during this period are suggested to explain the adaptation to drought, cold or disturbed environments observed in many Campanula species. Several independent long-distance dispersal events to North America are inferred within the Rapunculus clade, which seem to be related to high ploidy levels.

Molecular phylogeny ofCheirolophus (Asteraceae:Cardueae-Centaureinae) based on ITS sequences of nuclear ribosomal DNA

The genusCheirolophus has an interesting western Mediterranean and Macaronesian distribution. Here we investigate the delimitation of the genus and its exclusion from the large genusCentaurea, the systematic position of the related genusPaleocyanus, the delimitation of some species and the phylogeny of the group. We have carried out a phylogenetic analysis of the PCR-generated sequences of the internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA. The results suggest that the genus, includingPaleocyanus crassifolius is monophyletic; thus, a new combination of this species underCheirolophus is proposed. The Macaronesian group of species is also monophyletic, indicating a single colonization of the archipelago. The poor resolution of microspecies in the Macaronesian group reinforces the hypothesis of a very recent differentiation of the group.

Evolution and biogeography of Centaurea section Acrocentron inferred from nuclear and plastid DNA sequence analyses

BACKGROUND AND AIMS Section Acrocentron of the genus Centaurea is one of the largest sections of Centaurea with approx. 100 species. The geographic distribution, centred in the Mediterranean, makes it an excellent example for studies of the biogeographic history of this biodiversity-rich region. METHODS Plastid (trnH-psbA) and nuclear (ITS and ETS) DNA sequence analysis was used for phylogenetic reconstruction. Ancestral biogeographic patterns were inferred by dispersal-vicariance analysis (DIVA). KEY RESULTS The resulting phylogeny has implications for the sectional classification of Acrocentron and confirms merging sect. Chamaecyanus into Acrocentron as a subsection. Previous suggestions of an eastern Mediterranean origin of the group are confirmed. The main centres of diversification established in previous studies are now strongly supported. Expansion of the group in two different radiations that followed patently diverse paths is inferred. CONCLUSIONS Radiation followed two waves, widely separated in time scale. The oldest one, from Turkey to Greece and the northern Balkans and then to North Africa and Iberia, should be dated at the end of the Miocene in the Messinian period. It reached the Iberian Peninsula from the south, following a route that is landmarked by several relictic taxa in Sicily and North Africa. A later radiation during the Holocene interglacial periods followed, involving species from the north of the Balkan Peninsula, along a Eurasian pathway running from Central Iberia to the steppes of Kazakhstan. A generalized pattern of reticulation is also evident from the results, indicating past contacts between presently separated species. Molecular data also confirmed the extent of hybridization within Acrocentron and were successful in reconstructing the paleogeography of the section.

Delineation and phylogeny of Centaurea sect. Acrocentron based on DNA sequences: a restoration of the genus Crocodylium and indirect evidence of introgression

Abstract. The Mediterranean group Acrocentron of the genus Centaurea is defined mainly on the basis of pollen type, but also by achene characters and involucral bracts morphology. We have revised the delineation of the group by cladistically comparing the sequences of the ITS spacers of the nuclear ribosomal DNA. Our results confirm that the Acrocentron group is a natural one, with a different delimitation from the commonly accepted. The ITS phylogeny supports that Centaurea sect. Chamaecyanus and sect. Stephanochilus belong to the Acrocentron group and suggests that sect. Chamaecyanus should be merged in sect. Acrocentron as a subsection. Contrary, sect. Aegialophila and sect. Crocodylium form a natural group that cannot be placed in the Acrocentron group and should be considered a different genus. The inclusion of Centaurea crocodylium in Aegialophila makes that the prioritary name for the generic level is Crocodylium; thus, two new nomenclatural combinations are proposed: Crocodylium creticum and Crocodylium pumilum. The groups suggested by the ITS sequences are correlated to the main geographical centers of speciation of Acrocentron. However, support for internal nodes of the tree is extremely poor. The low support within the tree and the absence of correlation between karyology and molecular phylogeny suggest that hybridization has played an important role in the diversification of Acrocentron.

Phylogenetic relationships in the Mediterranean Helichrysum (Asteraceae, Gnaphalieae) based on nuclear rDNA ITS sequence data

The internal transcribed spacers ITS1 and ITS2 of the nuclear rDNA were sequenced for 41 Helichrysum species (Gnaphalieae), focusing on the Mediterranean group of species, together with eight representatives of other genera of the Gnaphalieae, in order to check the hypothesised monophyly of the Mediterranean Helichrysum group and the correspondence of the sequence data with its traditional sectional classification. The cladistic analysis of sequence data supports monophyly of the Mediterranean Helichrysum excluding H. frigidum and H. montelinasanum. The traditional classification of the Mediterranean species into two sections, Helichrysum and Virginea, is not supported, whereas a group constituted by species from the west Mediterranean area is shown as a moderately supported monophyletic clade in the strict consensus tree. Other results also show and confirm the complexity, still not satisfactorily resolved, of the Helichrysum generic delimitation: Pseudognaphalium luteoalbum appears merged in Helichrysum whereas Helichrysum dasyanthum appears more related to Anaxeton laeve than to any Helichrysum species.

Karyological evolution and molecular phylogeny in Macaronesian dendroid spurges (Euphorbia subsect. Pachycladae)

Abstract. Euphorbia subsect. Pachycladae is a taxon of primarily Macaronesian distribution, defined by morphological and biogeographical criteria. On the basis of morphological data, it is a heterogeneous group within which at least three complexes of species can be distinguished. To ascertain whether it is a natural group and discover its phylogenetic relations, we performed a cladistic analysis of the sequences of ribosomal nuclear DNA and a karyological study. The results of the two studies are concordant and show that the sub-section is polyphyletic and includes three different groups. The first monophyletic group is made up of the Macaronesian endemics E. atropurpurea complex and E. lamarckii complex, which form a polytomy with E. dendroides as the basal species. The lauroid species E. longifolia and E. stygiana represent the second monophyletic group, which derive from Mediterranean forms of E. sect. Helioscopia Dumort. Both species are paleopolyploid (2n=44) with highly symmetrical karyotypes. Finally, E. balsamifera, with a Canarian, African and Arabian distribution, remains isolated in a basal position. Its karyotype, with 2n=20 chromosomes, differs from the Macaronesian model and displays analogies with African cactiform spurges. On the basis of the results, some hypotheses are formulated about speciation processes in the three groups.

Allopolyploid origin of highly invasive Centaurea stoebe s.l. (Asteraceae)

Spotted knapweed (Centaurea stoebe) occurs from Western Asia to Western Europe both as diploid and tetraploid cytotypes, predominantly in single-cytotype populations with higher frequency of diploid populations. Interestingly, only tetraploids have been recorded so far from its introduced range in North America where they became highly invasive. We performed phylogenetic and network analyses of more than 40 accessions of the C. stoebe and C. paniculata groups and other related taxa using cloned internal transcribed spacer (ITS) and sequences of the chloroplast trnT-trnL and atpBrbcL regions to (i) assess the evolutionary origin of tetraploid C. stoebe s.l., and (ii) uncover the phylogeny of the C. stoebe group. Both issues have not been studied so far and thus remained controversial. Cloned ITS sequences showed the presence of two slightly divergent ribotypes occurring in tetraploid cytotype, while only one major ribotype was present in diploid C. stoebe s.str. This pattern suggests an allopolyploid origin of tetraploids with contribution of the diploid C. stoebe s.str. genome. Although we were not able to detect the second parental taxon, we hypothesize that hybridization might have triggered important changes in morphology and life history traits, which in turn may explain the colonization success of the tetraploid taxon. Bayesian relaxed clock estimations indicate a relatively recent--Pleistocene origin of the tetraploid C. stoebe s.l. Furthermore, our analyses showed a deep split between the C. paniculata and C. stoebe groups, and a young diversification of the taxa within the C. stoebe group. In contrast to nrDNA analyses, the observed pattern based on two cpDNA regions was inconclusive with respect to the origin and phylogeny of the studied taxa, most likely due to shared ancient polymorphism and frequent homoplasies.