Natural hybridization in Cardamine (Brassicaceae) in the Pyrenees: evidence from morphological and molecular data

While researching Cardamine (Brassicaceae) in the Pyrenees, putative hybrid plants were found at two natural sites. Pollen grain viability, AFLP, and multivariate morphometric analyses were performed in order to assess the plants' presumed hybrid origin, establishing that natural hybridization between the diploids C. crassifolia and C. amara ssp. pyrenaea had occurred. A new diploid nothospecies, C. × enriquei (2 n = 2x = 16), is described. Examination of 18 morphological characters showed the intermediacy of the hybrid between the parental taxa in most characters. AFLP analyses of C. amara ssp. amara , ssp. austriaca , ssp. olotensis and C. raphanifolia , demonstrated the close position of the hybrid to C. crassifolia , and revealed that the highest number of markers were shared with the parents. Polymorphism found in the AFLP pattern of the hybrid suggested recurrent origin, segregation and/or backcrosses, although assessment of pollen viability indicated high male sterility. The hybridization event reported here represents the second documented case between the C. pratensis group and C. amara . An account of the nomenclature of C. crassifolia is also presented, including lectotypification of relevant names.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 275–294.     

Hybridization between two polyploid Cardamine (Brassicaceae) species in North-western Spain: discordance between morphological and genetic variation patterns

BACKGROUND AND AIMS: Hybridization is an important evolutionary phenomenon, and therefore a detailed understanding of the dynamics of interspecific gene flow and resulting morphological and genetic patterns is of widespread interest. Here hybridization between the polyploids Cardamine pratensis and C. raphanifolia at four localities is explored. Using different types of data, the aim is to provide simultaneous and direct comparisons between genotype and phenotype variation patterns in the studied hybrid populations. METHODS: Evidence of hybridization has been gathered from morphology, molecular markers (amplified fragment length polymorphism and chloroplast DNA sequences), pollen viability, karyology and nuclear DNA content. KEY RESULTS: All data support extensive gene flow occurring in the hybrid populations. A wide range of morphological and genetic variation is observed, which includes both parental and intermediate types. Unbalanced pollen fertility and several ploidy levels are recorded. CONCLUSIONS: Incongruence reported between genotype and phenotype suggests that parental phenotypes are affected by introgression, and intermediate hybrid phenotypes can be genetically closer to one of the parents. Thus, it is evident that morphology, when used alone, can be misleading for interpreting hybridization, and critical evaluation of other data is needed.     

Cardamine apennina: a new endemic diploid species of the C. pratensis group (Brassicaceae) from Italy

Italian representatives of the Cardamine pratensis group were investigated using morphometric, karyological, and molecular (AFLP-fingerprinting) analyses. Focus was on resolving the taxonomic and evolutionary position of C. granulosa from Piemonte (NW Italy) and of populations from central Italy, previously included in C. granulosa or C. pratensis s. str. Results show that C. granulosa is a morphologically and genetically well defined diploid species, but is extremely rare and restricted to a small area in Piemonte. Central Italian populations differ from both C. granulosa and C. pratensis s. str., and, although the morphological differentiation is not strongly pronounced, they show marked molecular divergence, suggesting recognition as a new species, Cardamine apennina. Because of their restricted distribution and diploid chromosomal level, C. apennina and C. granulosa may represent basal and quite isolated lineages, whereas reticulation on diploid and polyploid level has affected remaining lineages of the C. pratensis group. Both Italian endemics are endangered due to rarity and habitat loss. A key for identification of the Italian taxa of the C. pratensis group is also provided.     

A new species of Cardamine subgen. Dentaria (Cruciferae), apoendemic in Calabria (Southern Italy)

ABSTRACT

Cardamine battagliae Cesca & Peruzzi sp. nova is related to Cardamine heptaphylla (Vill.) O. E. Schulz. (2n = 48). On the other hand, the new species is well distinguished both from a morphological and karyological point of view. Because of its distribution and high ploidy level (2n = ca. 160) C. battagliae can be considered as a new apoendemic taxon. Morphological and karyological data are presented and discussed.     

Towards a better understanding of the Chenopodium album aggregate (Amaranthaceae) in the Middle East: A karyological,cytometric and morphometric investigation

The study of variation in nuclear genome size, especially when combined with common garden experiments, significantly contributes to disentangling interspecies relationships within taxonomically complicated plant groups. The Chenopodium album aggregate is among the morphologically most variable groups and consists of many weakly differentiated cosmopolitan entities. We analysed nuclear genome size variation in diploid and polyploid species of the aggregate from Iran using flow cytometry of 282 accessions from 88 populations of 7 species. To this end, we also determined chromosome numbers and performed a morphometric study to reveal the extent of intraspecific morphological variation. We found that Iranian species are exclusively diploid (C. vulvaria), tetraploid (C. novopokrovskyanum, C. strictum, C. sosnowskyi and C. chaldoranicum) or hexaploid (C. album subsp. album, C. album subsp. iranicum and C. opulifolium). Six homogeneous relative genome size groups were distinguished among the species studied. Our morphometric study surprisingly revealed that under similar ecological conditions Chenopodium species are morphologically stable and well distinguishable, exhibited very little morphological variation. Hence, immense variation in leaf shapes, branching and inflorescence organization seen in the field has not been repeated under greenhouse conditions. The only exception was C. album s. str. which exhibited numerous morphotypes, covering the variation of remaining species.     

A new species of Haplophyllum A. Juss. (Rutaceae) from the Iberian Peninsula: evidence from morphological, karyological and molecular analyses

BACKGROUND AND AIMS: The discovery of a new species, Haplophyllum bastetanum F.B. Navarro, V.N. Suárez-Santiago & Blanca sp. nov., in the south-east of Spain has prompted the comparative study of species of the Iberian Peninsula, and others related, through morphological, cytogenetic, molecular, distributional and ecological characterization. METHODS: The morphological study involved a quantitative analysis of the species present in the Iberian Peninsula and a comparative analysis of the morphological characteristics between H. bastetanum and other related species. Mitotic analyses were made with root meristems taken from germinating seeds. Phylogenetic analyses of the internal transcribed spacer sequences of nuclear ribosomal DNA were performed using neighbour-joining (NJ) and maximum-parsimony methods. KEY RESULTS: Haplophyllum bastetanum is a diploid species (2n = 18) distinguished primarily for its non-trifoliate glabrous leaves, lanceolate sepals, dark-green petals with a dorsal band of hairs, and a highly hairy ovary with round-apex locules. The other two Iberian species (H. linifolium and H. rosmarinifolium) are tetraploid (2n = 36) and have yellow petals. Both phylogenetic methods generated a well-supported clade grouping H. linifolium with H. rosmarinifolium. In the NJ tree, the H. linifolium-H. rosmarinifolium clade is a sister group to H. bastetanum, while in the parsimony analysis this occurred only when the gaps were coded as a fifth base and the characters were reweighted according to the rescaled consistency index. This latter group is supported by the sequence divergence among taxa. CONCLUSIONS: The phylogenies established from DNA sequences together with morphological and cytogenetic analyses support the separation of H. bastetanum as a new species. The results suggest that the change in the number of chromosomes may be the key mechanism of speciation of the genus Haplophyllum in the Iberian Peninsula. An evolutionary scheme for them is propounded.     

A new species of Armeria (Plumbaginaceae) from southern Spain with molecular and morphometric evidence on its origin

The taxonomic status of the populations of Armeria from the Horconera massif (Cordoba province), previously misidentified or referred to without an explicit identification, is re-examined on the basis of morphological and molecular data. A multivariate analysis indicates that they can be discriminated from morphologically and geographically close taxa using morphometric characters. According to these results, and taking into account the previous evidence that supports extensive reticulation in the genus, a new species, A. trianoi , is formally described. Molecular evidence from our previous works, in particular the nuclear ribosomal ITS sequence data, provides the frame for a discussion on the origin of the new entity. It is suggested that A. trianoi may be the result of introgression of sympatric populations of A. villosa subsp. longiaristata into a pre-existing taxon, possibly A. filicaulis var. minor presently occurring in Sierra Tejeda some 50 km apart.     

Evolutionary history of the polyploid complex of Cardamine amara (Brassicaceae): isozyme evidence

 In samples from 56 populations of Cardamine amara, representing four diploid subspecies (subspp. amara, opicii, balcanica, and pyrenaea) and two tetraploid subspecies (subspp. austriaca and olotensis) from different parts of the European distribution area, four enzyme systems with 23 alleles were studied. These data, together with previous morphological and karyological data, suggest that the distribution and variation pattern within the species was strongly influenced by the last glacial period and postglacial migrations. Cardamine amara subsp. pyrenaea is monomorphic for a unique allele, and subsp. balcanica has a unique allele, too which, however, is not fixed in all populations of the taxon. Both taxa seem to be relic ones, although otherwise subsp. balcanica in respect of allelic spectrum much resembles subsp. amara. The other two diploid subspecies, subsp. amara and subsp. opicii, are not characterised by presence of unique alleles but differ in allele frequencies. The two tetraploid subspecies have different evolutionary histories. C. amara subsp. austriaca seems to be an autopolyploid derivative of subsp. amara which colonised open space offered by retreating glaciers in the Eastern Alps. C. amara subsp. olotensis from the Iberian Peninsula represents most probably a polyploid of preglacial time. Received June 22, 2001; accepted May 17, 2002 Published online: September 13, 2002     

Allopolyploid origin of Cardamine asarifolia (Brassicaceae): incongruence between plastid and nuclear ribosomal DNA sequences solved by a single-copy nuclear gene

Interspecific hybridization and polyploidization have played central roles in plant diversification. However, technical difficulties in the analyses of low-copy genes have limited the study of the origins of hybrid and polyploid plants. Here, we present a phylogenetic analysis of the hexaploid Cardamine asarifolia, distributed in the southern European Alps and northern Apennines. Our study included all relevant taxa of the genus found in Europe. A marked discrepancy was revealed between the trnL-trnF region of cpDNA and internal transcribed spacer (nrDNA ITS) sequences. To solve the incongruence, we sequenced a single-copy nuclear CHS gene (chalcone synthase) using a novel method to design homoeologue-specific PCR primers to bypass artefacts caused by artificial recombination of homoeologues during PCR and/or cloning. Three homoeologues were isolated from C. asarifolia, providing evidence for its allopolyploid origin. One homoeologue, showing the same phylogenetic position as the ITS sequences, most likely originated from an extinct parent. Furthermore, we documented recurrent polytopic hybridizations between C. asarifolia and diploid C. amara. The allohexaploidization and the following hybridization with a diploid species exemplify the ongoing dynamic processes of speciation in the genus Cardamine.     

Taxonomy and cytogeography of Cardamine raphanifolia and C. gallaecica (Brassicaceae) in the Iberian Peninsula

For a long time it was considered that Cardamine raphanifolia is widely distributed with several infraspecific taxa in the Mediterranean region. Recent studies showed that C. raphanifolia is restricted to the northern Iberian Peninsula. Cardamine gallaecica from NW Spain was regarded to be closely related, but was originally regarded as C. amara, and later described as C. raphanifolia subsp. gallaecica. In this study, we validate the treatment of C. raphanifolia and C. gallaecica as distinct species, and show their clear differentiation based on morphometric and molecular AFLP analyses. Moreover, both species are clearly distinct from C. amara subsp. pyrenaea, which has an overlapping distributional range with C. raphanifolia. Hexaploid (2n = 48) and octoploid cytotypes (2n = 64) of C. raphanifolia, and tetraploid (2n = 32) and hexaploid (2n = 48) cytotypes of C. gallaecica were found. Due to their weak morphological and genetic differentiation, these polyploid races are not classified as separate taxa.     

Contribution to the karyological knowledge of Echinops (Asteraceae, Cardueae) and related genera

Twenty-six chromosome counts were made of several genera of the tribe Cardueae from various European and Asian provenances: Acantholepis (one species studied), Amphoricarpus (1), Chardinia (1), Echinops (14 species, 15 populations), Siebera (1), Staehelina (3) and Xeranthemum (4). Eleven of the reports are made for the first time, ten confirm previous counts, while the remainder report disparities with earlier records. The existence of different basic chromosome numbers and ploidy levels suggests dysploidy and polyploidy as the main mechanisms of chromosomal evolution in the taxa considered.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 337−344.     

The Stegana (sensu stricto) species from China,with morphological and molecular evidence (Diptera: Drosophilidae)

Four new species of the subgenus Stegana (sensu stricto) were found and described from China: S. (S.) antha sp. nov., S. (S.) latiorificia sp. nov., S. (S.) huangjiai sp. nov. and S. (S.) nigrifoliacea sp. nov. Three known Stegana (s. str.) species from China: S. (S.) antlia Okada, 1991, S. (S.) cheni Sidorenko, 1997 Sidorenko, V. S. (1997). New Asian species and new records of the genus Stegana Meigen (Diptera, Drosophilidae). I. Subgenera Oxyphortica Duda and Stegana s. str. Annales d la Société Entomologique de France (N. S.), 33, 65–79.[Web of Science ®] , [Google Scholar] and S. (S.) emeiensis Sidorenko, 1997 Sidorenko, V. S. (1997). New Asian species and new records of the genus Stegana Meigen (Diptera, Drosophilidae). I. Subgenera Oxyphortica Duda and Stegana s. str. Annales d la Société Entomologique de France (N. S.), 33, 65–79.[Web of Science ®] , [Google Scholar] are redescribed as supplementary information. A key to the species, based on morphological characters, is provided. The phylogenetic relationships among 14 species of the subgenus Stegana are reconstructed with the combined sequences of the mitochondrial genes, ND2 (NADH dehydrogenase subunit 2) and COI (cytochrome c oxidase, subunit I). Moreover, the intra- and interspecific pairwise K-2P (Kimura's two-parameter) distances among these species are summarized, and the availability of mitochondrial markers in the species identification of the subgenus Stegana are further discussed.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:368C55C3-B101-457C-BD61-BDD8F1B780E5     

Distribution, chromosome numbers and nomenclature conspect of Arabidopsis halleri (Brassicaceae) in the Carpathians

Arabidopsis halleri represents an important model species for the study of phytoremediation. In the Carpathians it is represented by three subspecies: A. halleri subsp. halleri, A. halleri subsp. tatrica and A. halleri subsp. dacica. All three subspecies are diploid with chromosome number 2n = 16. They differ mainly in indument of flower parts, colour of petals and in the position of the longest leaf on stem. A. halleri subsp. halleri occurs in the Eastern and Southern Carpathians and in the northern and eastern part of the Western Carpathians, subsp. tatrica is endemic of the Western Carpathians and subsp. dacica occurs in the Eastern and Southern Carpathians most probably extending in its distribution further to the south to the Balkan mountains. Full synonymy of these three taxa and also a fourth European subspecies, A. halleri subsp. ovirensis is presented including the information on type specimens. Lectotypes are selected for several names. List of the examined herbarium specimens is given as well.     

Genetic and morphological variation in a Mediterranean glacial refugium: evidence from Italian pygmy shrews,Sorex minutus (Mammalia: Soricomorpha)

At the Last Glacial Maximum (LGM), the southern European peninsulas were important refugia for temperate species. Current genetic subdivision of species within these peninsulas may reflect past population subdivision at the LGM, as in ‘refugia within refugia’, and/or at other time periods. In the present study, we assess whether pygmy shrew populations from different regions within Italy are genetically and morphologically distinct. One maternally and two paternally inherited molecular markers (cytochrome b and Y‐chromosome introns, respectively) were analysed using several phylogenetic methods. A geometric morphometric analysis was performed on mandibles to evaluate size and shape variability between populations. Mandible shape was also explored with a functional approach that considered the mandible as a first‐order lever affecting bite force. We found genetically and morphologically distinct European, Italian, and southern Italian groups. Mandible size increased with decreasing latitude and southern Italian pygmy shrews exhibited mandibles with the strongest bite force. It is not clear whether or not the southern Italian and Italian groups of pygmy shrews occupied different refugia within the Italian peninsula at the LGM. It is likely, however, that geographic isolation earlier than the LGM on islands at the site of present‐day Calabria was important in generating the distinctive southern Italian group of pygmy shrews, and also the genetic groups in other small vertebrates that we review here. Calabria is an important hotspot for genetic diversity, and is worthy of conservation attention. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 774–787.     

Species delimitation in the Acomys cahirinus–dimidiatus complex (Rodentia, Muridae) inferred from chromosomal and morphological analyses

Our earlier chromosome banding studies of Acomys cahirinus and Acomys dimidiatus (the latter long considered to be a subspecies of the former) revealed that, despite very close diploid numbers (36 vs. 38), these taxa possess sharply different karyotypes and undoubtedly belong to different species. In this context, the taxonomic status and the relationship between the two chromosomal forms in Sinai (2 n  = 36) and Israel (2 n  = 38), chromosomally homozygous across a vast range except for a very narrow hybrid zone, remain poorly documented. Neither of these forms have previously been studied by chromosome banding; thus, the exact nature of chromosomal differences as well as the species to which these forms should be assigned remain unknown. Here, we present the data on comparative G-banding analysis and morphometrics of Acomys from Israel, Sinai, and Saudi Arabia, and a hybrid obtained in laboratory crosses between latter two. The analysis revealed that karyotype of Acomys from Israel is identical to that of Acomys from Saudi Arabia and both are different from that of Acomys from Sinai by one Robertsonian fusion. Therefore, karyotypically, all three are very different from A. cahirinus . It follows from the study that Sinai and probably Arabian peninsula and Minor Asia must be excluded from geographical distribution of A. cahirinus , which is limited from West Sahara to Egypt along Nile river (except Sinai). Furthermore, the synthesis of chromosomal and recent molecular data suggests a phylogeographical scenario explaining the modern distribution of Acomys in the Sinai and Arabian peninsulas and permits the update of the taxonomic status of these populations.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 203–214.     

Pleistocene refugia and recolonization routes in the southern Andes: insights from Hypochaeris palustris (Asteraceae, Lactuceae)

Hypochaeris palustris (Phil.) De Wild. is a species growing in the southern Andean chain. To elucidate potential Pleistocene refugia and recolonization routes in the southern Andes, we analysed amplified fragment length polymorphisms (AFLPs) in 206 individuals in 21 populations of H. palustris from the coastal Cordillera, the central, northern, and eastern ranges of the southern Andes, and Patagonia. Populations from the coastal Cordillera harboured more private AFLP fragments, and exhibited a higher frequency of polymorphic fragments as well as higher Shannon diversity than all other areas investigated. The comparison among pooled AFLP profiles of each region revealed that the central Andean ranges shared most fragments with populations from the margins of the distributional area in the Andes, in the N, E, and S (Patagonia). Phenetic analysis indicated close relationships among populations of the central ranges. Populations of the coastal Cordillera were shown to be highly differentiated from the Andean populations. It is very likely therefore that (1) H. palustris recolonized the central ranges of the southern Andes from nearby refugia, possibly unglaciated areas N, E, and/or S of its present distributional area; (2) the postglacial spread of H. palustris in the central ranges of the southern Andes occurred rapidly; and (3) the coastal Cordillera served as a refugium for H. palustris, but these populations did not contribute to the recolonization of the central Andean ranges.