Multiple diploid hybrid speciation of the Canary Island endemicArgyranthemum sundingii (Asteraceae)

There are several well-documented examples of multiple hybrid origins of polyploid species. Herein we report the first, to our knowledge, explicit example of a species that most probably has originated recurrently by diploid hybrid speciation. Genetic relationships and stabilization of twoArgyranthemum populations of putative hybrid origin on Tenerife, the Canary Islands, were investigated using chromosomal, morphometric, and fertility analyses of cultivated progeny families and artificial F₁ and F₂ hybrids. These data were compared to a recently published chloroplast DNA phylogeny of the genus, in which the same populations were included. The results suggest that the two populations must be referred to a single species,A. sundingii, which is diploid, fully fertile, genetically stabilized, and occurs in an ecologically intermediate habitat opened by deforestation, and that this species has originated at least twice following local hybridization in two valleys. The same parental species were involved in each origin; the montaneA. broussonetii and the coastalA. frutescens. The montane species was the chloroplast donor in one of the valleys and the coastal species in the other.     

The allopolyploid origin of Narcissus obsoletus (Alliaceae): identification of parental genomes by karyotype characterization and genomic in situ hybridization

Karyological and genomic in situ hybridization (GISH) approaches provided evidence of the parentage and origin of the hybrid species Narcissus obsoletus. Here, we demonstrate that the putative parental species, N. serotinus L. and N. elegans (Haw.) Spach, recently proposed because of their intermediate morphological traits, have participated in the hybridization process forming this taxon. Karyotype characterization of parental genomes in populations from S Spain and N Morocco has revealed differences in chromosome length and karyotype asymmetry, highlighting their diploid nature. Multicolour GISH on metaphase plates of N. obsoletus, with N. serotinus and N. elegans DNA used as probes, showed differential fluorescent staining of 10 and 20 chromosomes from parental genomes, respectively. Both parental genomes were detected in the allopolyploid, albeit in a duplicated manner. Secondary hybridization between N. obsoletus and N. serotinus was also detected karyologically. Little karyological differentiation between different geographic regions was found in either N. serotinus or N. obsoletus. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 477–498.     

Polyploidy and karyotype variation in Turkish Bellevalia (Hyacinthaceae)

There are 20 Bellevalia species in Turkey, half of which are endemic. Chromosome numbers are known for 15 species. A chromosome survey of 145 Bellevalia individuals showed that the karyotype is remarkably stable. All are based on x  = 4. The majority are diploid with 2 n  = 8, but there is also a polyploid series of 2 n  = 16, 24 and 32. Aneuploidy occurs only at the octoploid level. Eleven individuals had metacentric B chromosomes, one had acrocentric Bs and one had telocentric Bs. Bellevalia pycnantha and B. paradoxa are morphologically similar, with B. pycnantha reduced to a synonom of B. paradoxa .  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 87–98.     

Microsporogenesis and meiotic abnormalities in the hybrid complex of Phlomis composita (Lamiaceae)

The extent of chromosome structural differences such as permanent translocations and paracentric inversions has been studied in the hybrid complex of Phlomis composita and its putative parental taxa P. lychnitis and P. crinita . They are detected as meiotic abnormalities during microsporogenesis. In total, 117 plants from 19 populations in Andalusia (southern Spain) and Levante (eastern Spain) were studied for ploidy level, chromosome clumping, multivalent or univalent formation, late bivalent disjunction with bridges, chromatin bridges and fragments, irregular polysporads and variable pollen size. All individuals had the diploid chromosome number 2 n  = 20, with no significant difference in the frequencies of the above meiotic abnormalities either at taxon or population levels. However, very different frequencies were observed on the regional scale between populations in Andalusia and Levante. The results are discussed within the theoretical framework of homoploid hybrid or recombinational speciation.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 79−85.     

A new Pyrenean hybrid Cirsium (Asteraceae) as revealed by morphological and molecular analyses

A new nothosubspecies Cirsium  ×  vivantii is described after a molecular and morphological characterization of eight individuals collected in two separate valleys of the French central Pyrenees. Both hypervariable Amplified Fragment Length Polymorphism (AFLP) markers and nuclear rDNA (ITS) and plastid ( trn L-F, TRNT-L) DNA sequences were analysed. The profiles of these hybrid samples were compared to those of 43 individuals belonging to their presumed parental taxa C. carniolicum ssp. rufescens and C. palustre . A total of 133 AFLP bands were scored from three primer-pair combinations. All 130 AFLP bands that amplified in the hybrid samples were present in either C. carniolicum ssp. rufescens , C. palustre or both taxa, supporting the participation of these plant genomes in the resulting hybrids. Several Additive Polymorphic Sites (APS) detected in the ITS sequences of the hybrid samples also confirmed their derived origins from ribotypes of the two parental taxa. The lack of exclusive AFLP markers and the nonconcerted evolution of rDNA polymorphisms towards either of the parental ribotypes indicated their recent origin. Plastid trn T-L sequences were used to identify C. palustre as the maternal parent of six of these hybrid individuals; either parent could have acted as the plastid genome donor for the other two individuals. The morphological study revealed that all hybrid individuals were morphologically intermediate between their parents showing largely lobed and less spiny basal leaves as in C. carniolicum ssp. rufescens and decurrent leaf bases and pinkish corollas as in C. palustre .  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 421–434.     

Parnassia palustris: a genetically diverse species in Scandinavia

Patterns of variation at nine enzyme loci were examined in 528 plants representing diploid and tetraploid populations of Parnassia palustris s. l. in Europe to assess genetic variation patterns and migration history. Half of the plants showed a unique multilocus phenotype and 75% of all phenotypes occurred only in Scandinavia. Diploid populations showed similar levels of genetic diversity as other widespread outbreeding species with animal-mediated pollination and F -statistics indicated excessive heterozygosity and low rates of gene flow among them. In spite of dramatic population histories caused by the ice ages, diploid populations have maintained the same genetic diversity in Scandinavia as in central and southern Europe. Northern populations have apparently been established through the gradual advance of genetically variable populations and patterns of variation at individual loci indicate different migration routes, from the south-south-west and the east-north-east, respectively. The data strongly support a repeated autoploid origin of the tetraploid cytotype which has been much more successful than the diploid progenitors in colonizing new land since the last ice age. High genetic diversity in Scandinavia has apparently been obtained by a combination of immigration of plants from different source areas and recurrent formation of autotetraploids from diploid progenitors.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 347−372.     

Karyotype evolution in the genus Boronia (Rutaceae)

New chromosome counts are reported for Boronia clavata 2 n  = 14, B. heterophylla 'Near White' 2 n  = 15, B.  'Carousel' 2 n  = 16, B. deanei 2 n  = 22, B. chartacea 2 n  = 32, B. keysii 2 n  = 32, B. pilosa 2 n  = 44, B. anethifolia 2 n  = 36 and B. citriodora 2 n  = 108. Studies in 20 genotypes of 18 species and one interspecific hybrid revealed that they are highly complex in terms of chromosome number, ploidy level, chromosomal length, karyotype constitution and asymmetry. Karyotype analysis indicated that Boronia taxa with high chromosome numbers are primitive and those with lower numbers are derived. The basic chromosome number for this genus is suggested to be x = 18. Analysis of chromosome number, variations of total chromosome length (TCL) and average chromosome length (ACL), Nombre Fondamental (NF) and karyotype asymmetry suggest that dysploid reduction is the major mechanism in Boronia karyotype evolution. Chromosomal rearrangements might also have been involved. Origin, chromosome number changes and spread of Boronia are discussed in relation to the species divergence and the geological and climatic changes of the Australian continent.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 309–320.     

Isozyme variation among and within North Atlantic species of Papaver sect. Meconella (Papaveraceae) and taxonomic implications

The high-ploid species Papaver dahlianum , P. lapponicum , P. laestadianum and P. radicatum were compared by means of enzyme electrophoresis. The diploid P. alpinum from the Alps and garden material of the assumed diploid P. croceum from Asia were also included. The results are compared with those of previous morphological and cytological investigations. Generally, only small differences were found among the species. The maintenance of 13 subspecies of P. radicatum as published from the Nordic area was not supported. The octoploid P. laestadianum was nearly identical to the decaploid P. radicatum and inclusion in P. radicatum is justified. The octoploid P. lapponicum was also very similar to P. radicatum and inclusion of it in the latter should be considered. The most distinct of the taxa was the decaploid P. dahlianum , and the maintenance of two subspecies, ssp. polare and ssp. dahlianum can be defended. A hybrid origin of P. radicatum (10x) from P. lapponicum (8x) and P. alpinum (2x), and of P. dahlianum (10x) from P. lapponicum and P. croceum (2x) has been proposed, but a more likely explanation might be that P. dahlianum , P. radicatum and P. lapponicum have all arisen by polyploidization from closely related diploids without hybridization.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 255−269.     

Status and relationships of the extinct giant Canary Island lizard Gallotia goliath (Reptilia: Lacertidae), assessed using ancient mtDNA from its mummified remains

Ancient mitochondrial DNA sequences (378 base pairs of cytochrome b and 368 of 12S rRNA) extracted from a mummified extinct giant lizard, Gallotia goliath , from eastern Tenerife, Canary Islands, were used to assess the species status and relationship of this form within the genus. G. goliath is clearly a member of the G. simonyi group of the western Canary islands (Tenerife, La Gomera, El Hierro and La Palma) and is not closely related to the giant G. stehlini of Gran Canaria. Contrary to recent opinion, it is phylogenetically distinct, within the G. simonyi group, from the extant G. simonyi of El Hierro and also from the recently discovered live G. gomerana on La Gomera and from G. intermedia in north-western Tenerife. It may be the sister taxon of either all the other members of the G. simonyi group or of G. intermedia . The phylogenetic distinctness of G. goliath makes Tenerife unique among oceanic islands in having had one giant and two medium-sized lizard species that were probably substantially herbivorous, the others being G. intermedia and G. galloti . Gallotia shows great community differences on other islands in the Canaries, two having a single small species, one a single giant, and three a giant and a medium-sized form. © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 80 , 659–670.     

Uniformity of karyotypes in Ligularia (Asteraceae: Senecioneae), a highly diversified genus of the eastern Qinghai–Tibet Plateau highlands and adjacent areas

Ligularia , a highly diversified genus in the eastern Qinghai–Tibet Plateau and adjacent areas, was chosen as a suitable subject in which to study speciation patterns in this 'hot spot' area at the chromosomal level. Chromosome numbers and karyotypes were studied in 23 populations of 14 species, most of which are endemic to this area. The basic number x  = 29 was confirmed for all species. Ligularia virgaurea was found to have diploid and triploid cytotypes, 2 n  = 58 and 87. Other species are only diploid, with 2 n  = 58. The karyotypes of all populations within any species, and all species spanning most sections and covering most of the morphological range in Ligularia , are very similar to each other, belonging to type 2A according to Stebbin's classification. This karyotype was also found in its close allies, e.g. Cremanthodium , Ligulariopsis , Parasenecio , and Sinacalia . Aneuploid reduction of chromosome number from 2 n  = 60 to 58 and karyotypic variation was found in Ligularia and its allies. Such a chromosomal pattern with few polyploids infers that variation of karyotype structure at the diploid level seems to be the predominant feature of chromosomal evolution in this group and sympatric speciation via hybridization and polyploidization has played a minor role in its species diversity.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 329–342.     

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.     

Chromosome numbers in plant taxa endemic to the Balearic Islands

Mitotic chromosome numbers are reported from 20 vascular plant taxa that are endemic to the Balearic Islands and poorly known cytogenetically. The chromosome numbers of Arenaria grandiflora L. ssp. glabrescens (Willk.) G. López & Nieto Feliner (2 n  = 44), Dianthus rupicola Viv. ssp. bocchoriana L. Llorens & Gradaille (2 n  = 30), Solenopsis minuta (L.) C. Presl. ssp. balearica (E. Wimm.) Meikle (2 n  = 28), Romulea asumptionis Font Quer & Garcías Font (2 n  = 56), Scutellaria balearica Barceló (2 n  = 22) and Galium balearicum Briq. (2 n  = 22) have been determined for the first time. A new chromosome number was found in two populations of Helictotrichon crassifolium (Font Quer) Holub (2 n  =  c. 98) suggesting that this species is a high polyploid (14 x ), in contrast to an earlier report of a lower chromosome count (2 n  = 12 x  = 84). Cytogenetic observations suggest that Naufraga balearica Constance & Cannon has a diploid chromosome complement of 2 n  = 20, with 0–2 accessory chromosomes. The banded karyotype (chromomycin A₃) of Crepis triasii (Cambess.) Fries was determined for individuals belonging to eight populations from three islands. Two chromomycin A₃-positive regions were recorded in the diploid complement. These are present on the telomeric regions of the shortest subtelocentric chromosomes. This species is karyologically stable in chromosome number, karyotype and fluorochrome-banding pattern among populations from separate islands.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 219–228.     

Chromosome studies in Hippeastrum (Amaryllidaceae): variation in genome size

This paper presents the karyotype and DNA content of 12 diploid species of Hippeastrum from South America. The variation in genome size is compared with the karyotype and DNA content of Amaryllis belladonna from South Africa. The Hippeastrum species present a uniform and bimodal basic karyotype formula, but significant differences are found in the total chromosome volume (TCV) and nuclear DNA content. A positive correlation between the DNA content and TCV is also observed. The karyotype's constancy is a product of changes in DNA content occurring in the whole chromosome complement. The DNA addition to the long and short sets of chromosomes varies independently. In species with higher DNA contents, the short chromosomes add equal DNA amounts to both arms, maintaining their metacentric morphology, whereas the long chromosomes add DNA only to the short arm, increasing the chromosome symmetry. These data show that the evolutionary changes in DNA amount are proportional to chromosome length, maintaining the karyotypic uniformity. A. belladonna has a larger DNA content and possesses a karyotype different from that of Hippeastrum spp., supporting the distinction between the two genera and upholding the name Amaryllis for the South African entity against Hippeastrum for the South American genus.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 171–178.     

Molecular evidence for the hybrid origin of a new endemic species of Stylosanthes Sw. (Fabaceae) from the Mexican Yucatán Peninsula

Stylosanthes aff. calcicola is a formally undescribed tetraploid species from the Mexican Yucatán Peninsula, showing morphological similarities to the diploid species S. calcicola , but distinct in a number of characters. We used uni- and biparentally inherited molecular markers to infer the hybrid origin of this species in relation to known diploid species of Stylosanthes . Molecular characterization was based on length and/or DNA sequence variation of nuclear sequence-tagged site (STS) markers, the internal transcribed spacer (ITS) region of nuclear rDNA and the trnL intron of chloroplast DNA (cpDNA). Stylosanthes aff. calcicola contains a distinct cpDNA haplotype and nuclear DNA fragment, with closest relationship to the diploid species S. calcicola . In contrast, the DNA sequences of two nuclear loci reveal a closer relationship to the diploid species S. angustifolia , S. hispida , S. humilis , S. leiocarpa and S. viscosa . The majority of the STS markers showed additivity of PCR fragments in S. aff. calcicola , representing the combination of two genetically different genomes. We postulate that S. aff. calcicola is a distinct species of allotetraploid origin that appears to have originated once from hybridization between two divergent genomes, of which the maternal and paternal parent are closely related to, or derived from, a member of the lineages represented by S. calcicola and S. viscosa , respectively.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 140 , 1–13.     

Karyomorphology of Incarvillea (Bignoniaceae) and its implications in distribution and taxonomy

The karyomorphology of 11 species of the genus Incarvillea Juss. is reported. The chromosome numbers of all species studied are 2 n  = 22. The interphase nuclei and prophase chromosomes were found to be of the simple chromocentre type and the interstitial type, respectively. The asymmetry of the karyotype of I. arguta (two populations) in subgenus Amphicome is type 2A. The karyotypes of Incarvillea s inensis var. sinensis , I.  s inensis var. przewalskii, and I. olgae in subgenus Incarvillea are of asymmetry type 3A. The remaining nine species and one variety in subgenus Pteroscleris are also of asymmetry 3A. Data on three species and one variety studied are first reports. This study indicates that karyotype variation at the diploid level appears to be the predominant feature of chromosome evolution in the genus Incarvillea . According to this study of karyomorphology, morphological characteristics and geographical distribution, it seems that the three subgenera should be regarded as three independent genera. The geography of the genus is discussed.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 113–121.     

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.     

A molecular study of hybridization and homoploid hybrid speciation in Argyranthemum (Asteraceae) on Tenerife,the Canary Islands

Examples of recurrent homoploid hybrid speciation are few. One often‐cited example is Argyranthemum sundingii. This example includes two described species, A. lemsii and A. sundingii, resulting from reciprocal hybridization between A. broussonetii and A. frutescens on Tenerife. The four species and artificial F₁ and F₂ hybrids have previously been investigated morphologically and cytologically. Here, we examine population differentiation based on amplified fragment length polymorphism to get a better understanding of the genetic relationships among the species and the extent of hybridization. We aim to investigate if there is molecular support for treating the hybrid species as one taxon. Seven parental and four hybrid species populations (149 individuals) were analysed and we scored 85 polymorphic markers. A few (2–5) were private to each species but variably present and mostly rare. Our principal coordinate, STRUCTURE and BAPS analyses and AMOVA resulted in a clear separation of the parental species. The hybrid species were genetically less divergent but not identical. Our data indicate that hybridization and introgression are common in all these species on Tenerife and support the hypothesis that homoploid hybrid speciation has occurred repeatedly. Intrinsic post‐zygotic barriers are notoriously weak in Argyranthemum and reproductive isolation and speciation result primarily from strong ecological selection. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 19–31.     

Molecular evidence for repeated hybridization events involved in the origin of the genus × Crepidiastrixeris (Asteraceae) using RAPDs and ITS data

We investigated the hybrid origin of × Crepidiastrixeris denticulato-platyphylla using RAPDs and ITS sequence data. The putative parents Paraixeris denticulata and Crepidiastrum platyphyllum represent separate species, irrespective of geographical origin. The occurrence of species specific RAPD markers from P. denticulata and C. platyphyllum in × C. denticulato-platyphylla established unambiguously a hybrid origin between the two taxa. This was in line with the occurrence of a combination of morphological characters such as plant habit and floret numbers. The parent taxa differed from each other by 7 nucleotide substitutions and 2 indel events in the ITS region. The hybrids showed sequence additivity and most likely represent F₁ plants, with the exception of two plants which were of possible F₂ origin, possessing either the ITS sequences of one parent only, or one predominant ITS type. The hybrids occurred in two out of three localities where the parents occurred sympatrically. This fact, together with the short life-span of the plants, suggests that × C. denticulato-platyphylla exists as a result of repeated, frequent hybridization between the parent species.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 333–343.     

A new diploid Hieracium (Asteraceae: Lactuceae) from Bulgaria

Hieracium kittanae Vladimirov, a new diploid (2 n  = 18) species restricted to a few gorges in the Central Rhodope Mountains, south Bulgaria, is described and illustrated. It occurs in crevices of limestone rock in shady to sunny places with high atmospheric humidity. Morphologically, it to some extent resembles taxa from the H. glaucinum and H. schmidtii groups, but is otherwise very distinct, without close relatives in the Rhodope and neighbouring mountains.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003 , 143 , 213−218.