Morphological and molecular evidence of natural hybridization between two distantly related Rhododendron species from the Sino-Himalaya

Rhododendron (Ericaceae) is a large woody genus in which hybridization may play an important role in evolution and speciation, particularly in the Sino-Himalayan region, where many interfertile species often occur sympatrically. Natural hybridization between Rhododendron delavayi Franch. (=  R. arboreum ssp. delavayi ) and Rhododendron decorum Franch., which belong to different subsections of subgenus Hymenanthes, was investigated. Material of R. delavayi and R. decorum and their putative hybrids was collected from the wild. On the basis of morphology, chloroplast DNA, nuclear ribosomal DNA, and AFLP profiles, hybrids and parental species were identified. Hybridization occurred in both directions, but was asymmetrical, with R. delavayi as the major maternal parent in the hybrid zone. Most of the hybrids possessed intermediate phenotypes, and amongst the 15 hybrids detected were six F1s, two F2s, one first-generation backcross to R. delavayi , and two first-generation backcrosses to R. decorum . This indicates that, if Rhododendron underwent rapid radiation in this region, it did so in spite of permeable species barriers.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 156 , 119–129.     

Holcus azoricus M. Seq. & Castrov. (Poaceae), a new species from the Azores Islands

A new species of Holcus L. (Poaceae), endemic to the Azores archipelago (Portugal), is described. The vegetative and reproductive structures were studied. The diagnostic characters were compared with those of closely related species. New chromosome counts in Holcus azoricus M. Seq. & Castrov. (2 n  = 35) and H. rigidus Hochst. (2 n  = 28) are included. A possible hybrid origin is hypothesized; the potential parents are H. lanatus L. and H. rigidus .  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 259–267.     

Molecular and morphological evidence for hybridization between endemic Canary Island Convolvulus

Morphological data and molecular data from the chloroplast trn H- psb A region and nuclear ribosomal ITS region are used to test the hypothesis that the problematic Canary Island endemic Convolvulus floridus var. virgatus is a hybrid between the endemic species C. floridus and C. scoparius . Analysis of mean leaf length and width of 58 individuals indicates that accessions referable to C. floridus var. virgatus are intermediate between the parental taxa in leaf dimensions. Direct sequencing of the ITS region of C. scoparius and C. floridus revealed two species-specific ribotypes distinguished by 10 base differences. Examination of ITS chromatograms for putative hybrids revealed polymorphisms at those sites that are diagnostic between species in all except one putative hybrid. Morphological intermediacy and ITS additivity therefore support the hybrid status of C. floridus var. virgatus . An analysis of intraindividual ITS variation confirmed the co-occurrence of both parental ribotypes in putative hybrids and demonstrated that some hybrid individuals contained chimaeric ITS types. It is proposed that chimaeric ITS types are the result of recombination following backcrossing. Sequencing of the trn H- psb A region revealed four haplotypes. Three were, for the most part, confined to C. floridus and putative hybrids although one C. scoparius accession was also found to share this haplotype. The remaining haploype was only found in C. scoparius and putative hybrid accessions. Patterns of haplotype distribution between parental and hybrid accessions suggest multiple, bidirectional hybridization events between C. floridus and C. scoparius . The nomenclature of the hybrid is discussed and the necessary new name C.  ×  despreauxii is proposed.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 187–204.     

Morphological evidence for natural intergeneric hybridization in the New Zealand Gnaphalieae (Compositae): Anaphalioides bellidioides × Ewartia sinclairii

Wild intergeneric hybrids are recorded among six genera in the New Zealand Gnaphalieae, but little evidence substantiating their identity has been published. In this paper we evaluate the identity of putative hybrids between two species of Gnaphalieae endemic to New Zealand, Anaphalioides bellidioides and Ewartia sinclairii . The two species are occasionally sympatric in the north-east of the South Island and wild putative hybrids are rarely encountered. To test the hybridity hypothesis, specimens from nine putative hybrids were collected from one site and analysed for 76 morphological characters. The putative hybrids possessed predominantly intermediate or parental character states in comparison with A. bellidioides and E. sinclairii . Extreme traits were less common and a single novel character state was recorded in two hybrids only. Non-metric multidimensional scaling of dissimilarities derived from both continuous and discrete characters was strongly supportive of the hybridity hypothesis. Canonical discriminant analysis of the continuous characters was supportive for some putative hybrids only, in part reflecting the possession of extreme states in characters with high loadings. One of the putative hybrids was strongly indicated to be a backcross with A. bellidioides . The taxonomic treatment of cross-compatible genera is discussed.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 59−75.     

Genomic consequences of interspecific Pinus spp. hybridization

Strong karotypic orthoselection does not fully account for genome size variation in pines. Adult F1 interspecific pine hybrids are fertile and the genomic consequences of hybridization can be studied using haploid female megagametophytes. Greater variation in genome size was hypothesized to occur in hybrids compared to their parental species and the variation was thought to be positively related to the phylogenetic distance between the parental species. Nuclear 1C DNA content of megagametophytes from four sets of fertile Pinus spp. F1 hybrids and their parents was determined using a laser flow cytometer. Fertile F1 hybrids included two sets of hard pine hybrid Pinus elliottii Engelm. x P. caribaea var. hondurensis Morelet and two Asian x New World soft pine hybrids P. wallichiana A.B. Jacks. x P. strobus L. (hybrid is known as P. x schwerinii ) and P. lambertiana Dougl. x P. armandii Franchlet. Fertile, adult F1 produce haploid megagametophytes with nuclear DNA contents comparable to the parents or parental species. One genomic consequence of hybridization in pines is stability in nuclear DNA content. Hybrid genomes neither increased or decreased DNA content regardless of the phylogenetic distance between parents. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 75 , 503–508.     

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.     

Two new taxa and a new combination for Stipa (Gramineae: Stipeae) in Tunisia

Two new taxa in Stipa L. Sect. Leiostipa Dumort., S. alba and S. letournexii ssp. ignea , are described from southern Tunisia (northern Africa). In addition, the S. letournexii complex is reviewed and a new combination, S. letournexii ssp. tunetana (H.Scholz) F.M. Vázquez, is presented.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 439–444.     

Molecular and morphological characterization of Crataegus L. species (Rosaceae) in southern Syria

The systematics of the genus Crataegus (Rosaceae) have been considered problematic owing to the fact that hybridization, introgression, polyploidy and apomixis may occur in this genus. A study of the Crataegus species from the Arab mountains, Sweida Province, Syria, has been undertaken based on both plastid DNA sequences ( trn L- trn F, psb A- trn H) and morphological data. In the investigated region, three morphologically distinguishable Crataegus species: C. azarolus var. aronia L., C.  ×  sinaica Boiss. ssp. sinaica and C. monogyna var. monogyna Jacq. were investigated. Crataegus azarolus can be clearly distinguished morphologically from C. monogyna by the colour, size and structure of fruits, the number of pyrenes, the flowering and ripening time, the density of thorns, the tree shape and also the leaf shape. According to our morphological data, in Syria, C.  ×  sinaica is variable and could represent a hybrid of C. azarolus  ×  C. monogyna ; the cpDNA sequence analysis showed sequences corresponding to C. monogyna as the plausible mother of the hybrid.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 255–263.     

Genome organization in diploid hybrid species of Argyranthemum (Asteraceae) in the Canary Islands

Two diploid species of hybrid origin, Argyranthemum lemsii and A. sundingii , have been described from different valleys in the Anaga peninsula, north-east Tenerife. They have previously been shown to originate from hybridization between the same parental species, the montane A. broussonetii and the coastal A. frutescens , A. broussonetii being the chloroplast donor in one valley and A. frutescens in the other. The specific status of the two hybrid species has been questioned. In this study we used karyotype analysis, FISH, and GISH to address the question of multiple diploid hybrid speciation. GISH did not discriminate clearly between the parental genomes, but differential labelling was observed in separate hybrid populations, indicating different chromosomal rearrangements in different valleys. Small karyotype differences and local loss of rDNA were also observed. Thus separate origins of the same hybrid combination in different valleys in Tenerife have been verified. Our results add some support to the recognition of two species of hybrid origin, but the case serves to illustrate some of the many problems connected with the species concept in plants.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 141 , 491–501.     

Genetic structure and morphological variation of British populations of the hybrid Potamogeton x salicifolius

Potamogeton x salicifolius Wolfg. is one of the three most frequently recorded Potamogeton hybrids in the British Isles and Europe. It is thought to be the hybrid between P. lucens and P. perfoliatus. Its scattered distribution suggests that it has arisen several times in Britain. Most British populations of P. x salicifolius can be identified by their morphological characteristics, which are intermediate between those of the putative parents P. lucens and P. perfoliatus . However, the population at the Ouse Washes, Cambridgeshire, differs from other populations in its greater similarity to P . lucens . A genetic study of eight British populations, using six isozyme systems, revealed that most populations consist of a single multi-enzyme phenotype. This suggests that they were the result of a single hybridization event and are therefore maintained through vegetative reproduction. By contrast, the Ouse Washes population consists of three multi-enzyme phenotypes. This variation is likely to have resulted from multiple hybridization events, although we cannot exclude the possibility that the plants are partially fertile. The isozyme systems studied were unable to identify P. lucens and P. perfoliatus unambiguously, and consequently did not provide evidence for their putative parentage of P. x salicifolius. However, at a local level the banding patterns of the hybrids were generally consistent with the local multi-enzyme phenotypes of these putative parents.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 99–111.     

A peculiar new Orthotrichum species (Orthotrichaceae, Bryopsida) from central Argentina

A new epiphytic Orthotrichum species ( Orthotrichum spiculatum F. Lara, Garilleti & Mazimpaka sp. nov. ) is described from the sierra of Córdoba (Argentina). The new species has most of the characteristics of subgenus Pulchella (Schimp.) Vitt, but it is unique because of having eight exostome teeth pairs and 16 broad, strongly papillose endostome segments. Additionally, its upper leaves have green, acuminate, dentate–spiculate apices.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 477–482.     

Hybridization, developmental stability, and functionality of morphological traits in the ground beetle Carabus solieri (Coleoptera, Carabidae)

The assessment of developmental stability in hybrids can provide valuable information in the study of species formation because it allows an evaluation of the degree of incompatibility of genetic systems that control developmental processes. The present study assessed the impact of two hybridization events, assumed to have occurred at different times, on developmental instability in the ground beetle Carabus solieri . Developmental instability was estimated in 678 individuals from 27 populations from the fluctuating asymmetry (FA) levels of four morphological traits: the tibia length of middle and hind legs, which are functional structures, and the length and the proximal width of the hind wings, which are vestigial and thus nonfunctional structures. Significant variations of FA levels between populations were shown only for the wing width. For this trait, FA levels in hybrids were higher than in their parental entities for both hybridization events, indicating a significant divergence of the gene systems controlling development between the parental entities in the two hybridization cases. As expected, wing traits exhibited FA levels at least three times higher than leg trait. Finally, the potential interest of vestigial traits in the particular context of hybridization is discussed.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 151–158.     

Maintenance of species boundaries despite rampant hybridization between three species of reef fishes (Hexagrammidae): implications for the role of selection

Hybridization and introgression are antagonistic to the process of speciation. If hybrids are viable and backcross, introgression will slow or prevent the evolution of reproductive isolation between populations. If species hybridize upon secondary contact, introgression will ultimately erode species boundaries. If hybridization is rampant and gene flow is high, the proportion of hybrids will approach values expected with random mating, and introgression will overcome species boundaries, homogenizing populations. Alternatively, if species are genetically distinct, the proportion of hybrids would be significantly less than expected with random mating, gene flow would be interrupted by selection against hybrids, and inviability or sterility of F ₁ hybrids or backcrosses would be expected. Here, we investigate a system with characteristics expected by both of these opposing scenarios. Hybridization occurs between three species of reef fishes in the genus Hexagrammos at unexpected high frequencies in a zone of distributional overlap. Backcrossed individuals are detected, indicating F ₁ hybrids are viable and capable of reproducing. Yet, these species are genetically distinct at multiple loci. To study this apparent paradox, we estimate the relative proportions of hybrids, patterns of symmetry, inviability, and cytonuclear disequilibria using one mitochondrial and two nuclear markers. We invoke selection against hybrids, at various life history stages, in the maintenance of species boundaries in this system.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 135–147.     

Natural hybridization does not dissolve species boundaries in commercially important sea cucumbers

The study of species boundaries in areas of sympatry provides important insight into speciation processes. We investigated whether (i) two sympatric holothurians, Holothuria scabra and H. s. var. versicolor constituted species, and (ii) specimens of intermediate phenotype hybrids. Results from allozyme and 16S mtDNA sequence analyses indicated these two sea cucumbers to be distinct but young biological and phylogenetic species. Several private allozyme alleles existed and a Bayesian analysis grouped varieties into separate clusters. MtDNA sequences hardly varied within each taxon, and nine single bp changes were diagnostic between these two taxa. Allozyme allele frequencies in individuals of intermediate phenotype were intermediate to those of H. scabra and H. s.  var.  versicolor , most private alleles were present and heterozygote frequencies were higher than in either species. Ancestry coefficients modelled for these individuals were close to 0.5, indicating that the two taxa contributed equally to their genome. MtDNA sequences were identical to those of either species. We conclude that individuals of intermediate phenotype represent F1 hybrids. The presence of hybrids demonstrates that the opportunity for introgression exists, but is not realized, as backcrossing and introgression were not supported by the data. Thus, the genetic integrity of either holothurian species remains intact through an unknown postzygotic mechanism, possibly hybrid sterility.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 261–270.     

Non-metric morphological divergence in the western house mouse, Mus musculus domesticus, from the Barcelona chromosomal hybrid zone

The effect of hybridization on morphological variation was investigated in 120 western house mice, Mus musculus domesticus , from the hybrid zone between the Barcelona and standard chromosomal races. The incidence of 37 non-metric cranial traits was calculated for standard mice (2 n  = 40) and Barcelona-standard hybrids (2 n  = 27–39). Subsequent analyses were conducted on several karyological subgroups, established by grouping the animals according to either their diploid number or their degree of chromosomal heterozygosity. Results revealed no significant difference by sex, asymmetry, or geographical distance. Significant phenetic divergences were found between the karyotypes studied in relation to several variants. Differences were especially substantial between the standard race and hybrid mice, even with respect to those hybrids with karyotypes close to that of the standard race. Within the hybrids, the maximum divergence corresponded to the 28-chromosome homozygotes, chromosomally close to the Barcelona race, and to the heterozygotes with more than two fusions. Since differences in non-metric trait frequencies are generally considered a measure of genetic divergence, the results suggest the occurrence of a barrier to gene flow in the Barcelona hybrid zone. The decrease of genetic exchange between the chromosomally differentiated mice might be due to reduced fertility in hybrids, associated with chromosomal heterozygosity.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 80 , 313–322.     

Ferula mervynii (Apiaceae), a distinct new species from north-east Anatolia, Turkey

Ferula mervynii M. Sağıroğlu & H.Duman, sp. nov. from north-east Anatolia in Turkey (A8: Artvin & Erzurum) is described and illustrated. It is closely related to F. coskunii H.Duman & M.Sağıroğlu, from which it differs mainly in its habit, basal leaves, umbel, and mericarp features. Diagnostic morphological characters are discussed. Notes are also presented on its ecology and conservation status, together with scanning electron micrographs of the surface of the mericarps and of a somatic metaphase plate showing the chromosome number.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 357–362.     

A new species of Ceratozamia (Zamiaceae) from the Sierra Madre of Chiapas, Mexico, with comments on species relationships

The new species Ceratozamia vovidesii from a Pleistocene floristic refuge in southern Mexico is described and illustrated. It show an affinity with C. matudae Lundell and C. mirandae Vovides, Pérez-Farrera & Iglesias from Chiapas, but it differs from them in leaf, male and female cones, and trunk morphology.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 393–400.     

Euphrasia amurensis, E. hirtella var. ramosa and var. karoiana (Orobanchaceae)

The lectotype for Euphrasia hirtella var. ramosa is selected. This taxon and E. hirtella var. karoiana are compared with E. amurensis and confirmed as synonyms, corroborating Juzepcuk (1955 ).  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 215–223.     

Relationships in Patagonian species of Berberis (Berberidaceae) based on the characterization of rDNA internal transcribed spacer sequences

Sequence analysis of the internal transcribed spacer (ITS) of the 18S(ITS1)-5.8S-26S(ITS2) rDNA region was performed in order to analyse the phylogenetic relationships between 13 Patagonian species of the genus Berberis (Berberidaceae). The divergence values between the pairwise sequence in the studied Patagonian species were in the range 2.9–22.9%. The lengths of the ITS1 and ITS2 sequences were in the range 227–231 bp and 220–224 bp, respectively, and the 5.8S sequence was 159 bp throughout all species . B. microphylla sensu Landrum does not appear to be monophyletic based on current sampling. Indeed, we suggest that B. microphylla should be distinguished from B. buxifolia , B. parodii , and B. heterophylla . ITS sequences, together with data obtained from morphological, biochemical, amplified fragment length polymorphism, and cytological characterizations, support the existence of diploid and polyploid hybrid speciation in the genus.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 321–328.