Evolutionary patterns in the antR-Cor gene in the dwarf dogwood complex (Cornus, Cornaceae)

The evolutionary pattern of the myc-like anthocyanin regulatory gene antR-Cor was examined in the dwarf dogwood species complex (Cornus Subgenus Arctocrania) that contains two diploid species (C. canadensis and C. suecica), their putative hybrids with intermediate phenotypes, and a tetraploid derivative (C. unalaschkensis). Full-length sequences of this gene (∼4 kb) were sequenced and characterized for 47 dwarf dogwood samples representing all taxa categories from 43 sites in the Pacific Northwest. Analysis of nucleotide diversity indicated departures from neutral evolution, due most likely to local population structure. Neighbor-joining and haplotype network analyses show that sequences from the tetraploid and diploid intermediates are much more strongly diverged from C. suecica than from C. canadensis, and that the intermediate phenotypes may represent an ancestral group to C. canadensis rather than interspecific hybrids. Seven amino acid mutations that are potentially linked to myc-like anthocyanin regulatory gene function correlate with petal colors differences that characterize the divergence between two diploid species and the tetraploid species in this complex. The evidence provides a working hypothesis for testing the role of the gene in speciation and its link to the petal coloration. Sequencing and analysis of additional nuclear genes will be necessary to resolve questions about the evolution of the dwarf dogwood complex.     

Upslope movements and large scale expansions: the taxonomy and biogeography of the Coenonympha arcania –C. d arwiniana –C. gardetta butterfly species complex

Sibling species groups are suitable models for the understanding of inter‐ and intraspecific processes in taxonomy and biogeography. We analysed 262 individuals from the Alps of the Coenonympha arcania/gardetta species complex by allozyme electrophoresis. These taxa showed high variance amongst populations (F_ST: 0.391) and strong intertaxon genetic differentiation (F_CT: 0.376). Although morphologically similar, Coenonympha gardetta and Coenonympha arcania clearly differ in their genetic characteristics; the morphologically intermediate taxa Coenonympha darwiniana darwiniana and Coenonympha darwiniana macromma are genetically well distinguished from each other and the two other taxa. Coenonympha arcania and C. d. macromma most probably share a common ancestor and evolved by cladogenesis, whereas the taxonomic situation of C. d. darwiniana is still unresolved: This taxon might be the result of hybridization between C. arcania and C. gardetta or it might have a common ancestor together with C. gardetta. We suggest species rank for all four taxa. The distribution of genetic diversity of these populations and the differentiation amongst populations suggest rather different biogeographical scenarios: C. arcania most probably is of Mediterranean origin with postglacial range expansion northwards; C. gardetta survived the last ice age in peripheral refugia of the Alps and has spread all over this high mountain system in the postglacial; C. darwiniana and C. macromma survived the Würm in geographic proximity to their actual distribution areas and only have performed moderate uphill translocations during postglacial warming. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159 , 890–904.     

Independent origins of tetraploid cryptic species in the fern <Emphasis Type="Italic">Ceratopteris thalictroides</Emphasis>

Ceratopteris thalictroides (L.) Brongn is a tetraploid fern species that contains at least three cryptic species, the south, the north and the third type. In this study we combined data from both chloroplast DNA (cpDNA) and nuclear DNA sequences of three diploid species and three cryptic species of C. thalictroides to unravel the origin of the cryptic species, particularly of the reticulate relationships among the diploid and tetraploid taxa in the genus Ceratopteris. Of the three diploid species examined, C. cornuta had cpDNA identical to that of the tetraploid third type plants, and this diploid species is a possible maternal ancestor of the tetraploid third type. Analysis of the homologue of the Arabidopsis thaliana LEAFY gene (CLFY1) identified ten alleles in the genus Ceratopteris, with six alleles found in C. thalictroides. The unrooted tree of the CLFY1 gene revealed four clusters. Each cryptic species showed fixed heterozygosity at the CLFY1 locus and had two alleles from different clusters of the CLFY1 tree. Consideration of the cpDNA sequences, CLFY1 genotypes of the cryptic species and CLFY1 gene tree in concert suggested that the cryptic species of C. thalictroides had originated through independent allopolyploidization events involving C. cornuta and two unknown hypothetical diploid species.     

Chromosome numbers, karyotypes and nuclear DNA contents from perennial polyploid groups ofCerastium (Caryophyllaceae)

Somatic chromosome numbers have been determined for the followingCerastium taxa:C. eriophorum (2n = 36),C. alpinum (2n = 72),C. transsylvanicum (2n = 108),C. arcticum (2n = 108),C. latifolium (2n = 36),C. carinthiacum (2n = 36),C. banaticum (2n = 36),C. arvense subsp.glandulosum (2n = 36),C. arvense subsp.arvense (2n = 72) andC. fontanum (2n = 144). Karyotypes of three diploid species (C. eriophorum, C. banaticum andC. latifolium), belonging to three different taxonomic groups, were analysed and found to be similar. The relative nuclear DNA contents of all taxa were determined by flow cytometry and, for five species, also by Feulgen cytophotometry. The values obtained by the two methods are similar. A comparison of nuclear DNA contents among diploids shows that values differ significantly between different taxonomic groups, and are correlated with average chromosome size. Within closely related polyploid groups nuclear DNA amounts increase from 2x- to 4x- and 6x taxa as 1 : 1.4 : 2.4 in theC. alpinum complex, whereas DNA amounts are doubled comparing 2x- and 4x-subspecies in theC. arvense complex.     

Cytotaxonomy and breeding system of the genusBiscutella (Cruciferae)

Chromosome counts were determined for 46 populations ofBiscutella representing 28 taxa. The genus was found to contain diploid taxa with 2n = 12, 16 and 18, tetraploid taxa with 2n = 36 and hexaploid taxa having 2n = 54.B. laevigata L. s. l. consists of diploid and tetraploid populations which are poorly differentiated morphologically. TetraploidB. laevigata s. l. and hexaploidB. variegata Boiss. & Reuter (s. l.) are characterized by chromosomal instability. The variation in chromosome numbers and the occurrence of polyploidy is discussed in relation to the taxonomy of the genus. An investigation of the breeding system showed that most of the annual species were self-compatible and partly inbreeding and most of the perennial species self-incompatible and, therefore, outbreeding, while one annual species,B. cichoriifolia Loisel., showed both systems.     

Polyploid evolution and biogeography in Chelone (Scrophulariaceae): morphological and isozyme evidence

Chelone is a genus of perennial herbs comprising three diploid species (C. cuthbertii, C. glabra, and C. lyonii) and a fourth species (C. obliqua) that occurs as tetraploid and hexaploid races. To assess patterns of isozyme and morphological variation, and to test hypotheses of hybridization and allopolyploidy, we analyzed variation among 16 isozyme loci from 61 populations and 16 morphological characters from 33 populations representing all taxa and ploidy levels. Based on morphological analyses using clustering (unweighted pair group method using an arithmetic average) and ordination (principal components analysis and canonical variance analysis) methods, we recognize three diploid species without infraspecific taxa. Polyploids in the C. obliqua complex were most similar morphologically to diploid populations of C. glabra and C. lyonii. Patterns of isozyme variation among polyploids, which included fixed heterozygosity and recombinant profiles of alleles present in diploids, suggested polytopic origins of tetraploids and hexaploids. Our data indicate independent origins of polyploids in or near the southern Blue Ridge, Interior Highlands and Plains, and Atlantic Coastal Plain regions from progenitors most similar to C. glabra and C. lyonii. Extant tetraploids were not implicated in evolution of hexaploids, and plants similar to C. cuthbertii appeared unlikely as diploid progenitors for polyploids. We propose multiple differentiation and hybridization/polyploidization cycles in different geographic regions to explain the pattern of allopatry and inferred polytopic origins among polyploids.     

Karyosystematic studies on threeCrepis species (Asteraceae) endemic to Greece

This work examines the cytogeographical distribution, the morphological characters, and the karyotypes of threeCrepis species endemic to Greece (C. sibthorpiana, C. incana, andC. heldreichiana). C. sibthorpiana is diploid (2n = 2x = 8),C. incana is diploid (2n = 2x = 8) and tetraploid (2n = 4x = 16, 17), andC. heldreichiana is always dekaploid (2n = 10x = 40). The Giemsa positive bands, usually pairs of dots, are mainly centromeric inC. incana, while they are terminal inC. sibthorpiana (on the short arm of all chromosomes) and inC. heldreichiana (on both arms of all chromosomes). Intercalary C-bands are scarce and usually variable within karyotypes, individuals, and species. The most variable karyotype both in Feulgen and Giemsa preparations is that ofC. incana, in which also supernumerary chromosomes were observed, which are polysomic to standard set members. On the basis of morphological and karyological data the evolutionary relationships among the threeCrepis taxa are discussed.     

Reticulate evolution and phenotypic diversity in North American ciscoes, Coregonus ssp. (Teleostei: Salmonidae): implications for the conservation of an evolutionary legacy

Coregonine fishes are notorious taxonomicproblems due to their extreme morphological andecological variation. In North America, diversity is particularly baffling among ciscoes, and both morphological and phylogenetic analyses have resulted in major polytomy among the 8 taxa of the ``Coregonus artedi' species complex. Ciscoes arealso a devastated group, accounting for 10% ofthe fish species listed by the Committee on theStatus of Endangered Wildlife in Canada. Here,we complete the genetic characterization ofNorth American ciscoes with mitochondrial andmicrosatellites markers previously used toanalyse populations of C. artedi in orderto elucidate the evolutionary history andidentify appropriate conservation units. Ourresults revealed a complex evolutionary historymarked by postglacial reticulation eventscoupled with recent and independent evolutionof similar phenotypes (taxa). Genetic variationreflects geography rather than taxonomy, andconsequently, we recommend that a single taxon,C. artedi (sensu lato) be recognized.Local genetic differentiation is often coupledwith ecophenotypic diversification, and gillraker polymorphisms, depth-related habitatpreference and reproductive behaviour areconsidered as phenotypic traits with probableadaptive value contributing to the nicheexpansion of ciscoes. Ecomorphotypes of eachparticular locale thus represent a uniqueexpression of a diverse genetic pool stillundergoing divergence and sorting.Consequently, ciscoes from lakes with distinctecomorphotypes are recognized as ESUs, as wellas each of sympatric forms when they aregenetically differentiated. We recommend thatan ESU strategy focusing at a very local levelbe adopted for continental ciscoes as a validalternative to protect significant evolutionaryprocesses of divergence encountered inpolytypic species of newly colonized habitats.     

Genetics,taxonomy and the conservation of British <Emphasis Type="Italic">Euphrasia</Emphasis>

In Britain the genus Euphrasia comprises ca 20 diploid and tetraploid plant species, including several endemics. However, their conservation is impeded by taxonomic uncertainty. Analysis of cpDNA and AFLP variation was used to assess their taxonomic status and establish the extent of barriers to gene exchange among them. Differences in ploidy level constitute a very strong barrier to genetic exchange, although this is not absolute. The diploid endemics E. vigursii and E. rivularis form morphologically and genetically definable units which show some level of reproductive isolation. Within tetraploid Euphrasia, the species showed varying degrees of distinctness. Analysis of geographically paired samples from two widespread outcrossing taxa E. arctica and E. nemorosa provides evidence for extensive genetic exchange between them. However AFLP data indicate that this outbreeding species complex possesses a gene pool distinct from that of the widespread inbreeding tetraploids. The widespread and endemic inbreeding tetraploids contain examples of morphologically and genetically definable taxa, but also species whose distinctness is more equivocal. The conservation implications of this study are that species-based action plans are potentially suitable for conservation of the diploid endemics E. vigursii and E. rivularis. In contrast we contend that a species-based conservation framework, developed with reproductively isolated and genetically distinct groups in mind, requires modification for conservation of the complex and dynamic diversity found within the tetraploids. The adoption of ‘taxonomic’ action plans, designed to protect the evolutionary processes generating Euphrasia diversity, may provide a supplementary solution for conserving this type of variation.     

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.     

Chloroplast DNA and isozyme evidence on the evolution ofSenecio vulgaris (Asteraceae)

Chloroplast DNA (cpDNA) and isozyme variation were analyzed over a range of populations of two infraspecific taxa of the tetraploidSenecio vulgaris. The isozyme data were supportive of the hypothesis that the weedy and cosmopolitanS. vulgaris var.vulgaris is an evolutionary derivative ofS. vulgaris subsp.denticulatus from the coasts of W Europe and montane altitudes in S Spain and Sicily. The two taxa exhibited a very high genetic identity with subsp.denticulatus containing slightly more isozyme diversity than was found in var.vulgaris. — Three cpDNA haplotypes (A, B, C) already known from other Mediterranean diploid species ofSenecio were resolved in var.vulgaris, and an additional fourth haplotype (E) was found in subsp.denticulatus. Two alternative hypotheses were chosen to account for the origin and maintenance of the observed cpDNA composition ofS. vulgaris. It either reflects (1) the retention of an ancestral polymorphism which stems from the recurrent and polytopic formation of ancestral tetraploid lineages; or (2)S. vulgaris originally was characterized by haplotype E, and haplotypes A, B and C were acquired through repeated introgressive hybridization with related diploid species. The finding that very low levels of nuclear (isozyme) diversity were present in both taxa ofS. vulgaris examined supports the second of these two hypotheses; however, more detailed analysis of nuclear genetic diversity is required before a firm conclusion can be reached on this matter.Dedicated to emer. Univ.-Prof. DrFriedrich Ehrendorfer on the occasion of his 70th birthday     

A CHEMOSYSTEMATIC STUDY OF SELECTED TAXA OF CAPSICUM

Six taxa of Capsicum were chosen for a comparative chemosystematic study. A “key” individual from each taxon was selected for intensive chemical investigation. Thirteen flavonoids were isolated from leaf material and characterized by paper chromatography and absorption spectroscopy. The C-glycosylflavones vitexin and isovitexin, and orientin and iso-orientin, as well as O-glycosides of the flavones apigenin, luteolin, and chrysoeriol, were isolated from the key individuals. Chromatographic analysis of collections from various regions of South America, Central America, and Mexico showed in general that flavonoid variability is more common in cultivated taxa than in wild. Three groups of Capsicum were recognized and the main systematic conclusions were: (1) The white-flowered taxa in Group I, C. baccatum var. baccatum and C. baccatum var. pendulum, have identical flavonoids, corroborating previous conclusions that they are one species. The absence of chrysoeriol in this group separates it from Groups II and III. (2) The purple-to-white-flowered C. eximium var. eximium and C. eximium var. lomenlosum, Group II, have a complex flavonoid chemistry which appears to link Groups I and III. (3) Two purple-flowered species, C. cardenasii and C. pubescens, Group III, are chemically distinct from the other taxa examined.     

Distinction in morphology and esterase isozyme betweenEupatorium glehni (=E. chinense subsp.sachalinense) andE. chinense var.oppositifolium (compositae)

The taxonomic status ofEupatorium chinese var.oppositofolium andE. glehni (=E. chinense subsp.sachalinense) inE. chinense complex semsu Kitamura has long been controversial. In this paper, the degree of divergence between diploids of these two taxa was examined by means of morphological studies including principal component analysis, the electrophoretic analysis of esterase isozyme variation and observations on habitats. The data obtained through the examinations indicate these two taxa are diverged enough to be recognized as distinct biological species. Since the polyploidE. chinense var.oppositifolium is more or less intermediate in morphology between the two diploid taxa, it is considered to have masked the distinction between the two diploid taxa. Also, electrophoretic evidence suggests that polyploidE. chinense var.oppositifolium is not a hybrid or hybrid derivative withE. glehni as a parental species. Possible origin of polyploidE. chinense varoppositifolium is also discussed.     

Nuclear DNA content in the genus Hepatica (Ranunculaceae)

Using flow cytometry, we measured the nuclear DNA contents of all known taxa in Hepatica. Nuclear DNA content of Hepatica falconeri (diploid, crenate leaf lobes) was significantly lower than that of diploid entire species. Among the tetraploid species, crenate species had lower DNA contents than the entire taxon H. nobilis var. pubescens. The DNA content of the tetraploid species was more than double that of the diploid species among the same leaf-type groups.     

Allozyme variation in Japanese species ofChionographis (Liliaceae)

Allozyme variation was examined in three diploid taxaChionographis japonica var.japonica, var.kurokamiana, andC. koidzumiana and three tetraploid taxaC. japonica var.kurohimensis, ssp.hisauchiana, and ssp.minoensis. Results show thatC. japonica var.kurokamiana is genetically closer toC. koidzumiana than to var.japonica. In the tetraploid taxa, fixed heterozygosities were found at several loci, and this supports the hypothesis that these taxa are allotetraploids. Furthermore, the tetraploid taxa have many unique alleles not found in the diploid taxa. This suggests that sufficient time has passed since the origin of tetraploids for new mutations to have been fixed.     

Exudate flavones and flavanones in Origanum species and their interspecific variation

Surface flavonoids in nine species of Origanum, representing taxa from all three of the currently recognised subgeneric groups, were examined both by HPLC coupled to diode-array detection and APCI–MS. Many of the flavonoids present were characterised by O-substituent at C-6 (OH, OMe) and/or C-8 (OMe). In total, 25 flavones and flavanones are described in this study, of which 13 are new to the genus and 5,4′-dihydroxy-6,7,3′-trimethoxyflavanone is reported for the first time. Taxa in subgeneric Group A accumulated flavonoids with methoxyl groups at both C-6 and C-4′; however, taxa in subgeneric Group B did not accumulate 4′-methoxylated compounds, and taxa in Group C did not accumulate 6-methoxylated compounds.     

Genetic differentiation and speciation in prealpine Cochlearia: Allohexaploid Cochlearia bavarica Vogt (Brassicaceae) compared to its diploid ancestor Cochlearia pyrenaica DC. in Germany and Austria

 Significant geographic partitioning of genetic variation within Cochlearia bavarica was found within populations from Allg?u and SE Bavaria (Germany) exhibiting significant genetic differentiation. It has been demonstrated that allohexaploid C. bavarica evolved via hybridization between diploid C. pyrenaica and tetraploid C. officinalis. Presently, only C. pyrenaica is distributed throughout inland Central Europe. It has been concluded that C. bavarica is of inter- or postglacial origin, and its speciation was not influenced by human activities. Isozyme analysis revealed that there is a correlation between interpopulational genetic distances and geographic distances among C. bavarica populations from both regions, and which is not the case for C. pyrenaica in Germany and Austria. Only high alpine C. excelsa is significantly differentiated among the diploid taxa analysed here. Geographically structured distribution of alleles and their frequencies in C. bavarica populations could not be explained with the distribution of these alleles in C. pyrenaica. The presented findings favour disruption of a former wider distribution area rather than migration of C. bavarica or a polytopic origin of this species. Received April 17, 2001 Accepted February 1, 2002     

Chloroplast DNA variation in diploid and polyploid species of Bromus (Poaceae) subgenera Festucaria and Ceratochloa

Summary Chloroplast DNA (cpDNA) restriction endonuclease patterns are used to examine phylogenetic relationships between Bromus subgenera Festucaria and Ceratochloa. Festucaria is considered monophyletic based on the L genome, while Ceratochloa encompasses two species complexes: the B. catharticus complex, which evolved by combining three different genomes, and the B. carinatus complex, which is thought to have originated from hybridization between polyploid species of B. catharticus and diploid members of Festucaria. All species of subgenus Ceratochloa (hexaploids and octoploids) were identical in chloroplast DNA sequences. Similarly, polyploid species of subgenus Festucaria, except for B. auleticus, were identical in cpDNA sequences. In contrast, diploid species of subgenus Festucaria showed various degrees of nucleotide sequence divergence. Species of subgenus Ceratochloa appeared monophyletic and phylogenetically closely related to the diploid B. anomalus and B. auleticus of subgenus Festucaria. The remaining diploid and polyploid species of subgenus Festucaria appeared in a distinct grouping. The study suggests that the B. catharticus complex must have been the maternal parent in the proposed hybrid origin of B. carinatus complex. Although there is no direct evidence for the paternal parent of the latter complex, the cpDNA study shows the complex to be phylogenetically very related to the diploid B. anomalus of subgenus Festucaria.     

Cochlearia macrorrhiza (Brassicaceae): A bridging species between Cochlearia taxa from the Eastern Alps and the Carpathians?

Cochlearia macrorrhiza is one of the most highly endangered species in Central Europe and less than five individuals survived at its natural stand in a lowland area between the Eastern Alps and the Carpathians. Amplified fragment length polymorphisms (AFLPs) confirmed the status of C. macrorrhiza as a distinct taxon. Lowland C. macrorrhiza does not bridge the distribution of montainous and alpine Cochlearia species from the Eastern Alps and the Carpathians genetically, and C. macrorrhiza represents a separate lineage which evolved from diploid Cochlearia as C. excelsa in East Austrian high alpine regions did. Another species considered in this study, the Romanian C. borzaeana is more closely related to C. tatrae from the High Tatra mountains than to C. pyrenaica from Slovakia or Austria and the AFLP results suggest a single origin of alpine 2n=42 taxa. Genetic differentiation within and between populations is highly structured geographically, and the AFLP data favour a former widespread distribution of C. pyrenaica in mountainous regions and a parallel evolution of high alpine taxa in the Eastern Alps and the Carpathians, respectively.     

The phylogenetic relationships of Cynoglottis (Boraginaceae- Boragineae) inferred from ITS, 5.8S and trnL sequences

A molecular phylogenetic analysis of Cynoglottis was performed to evaluate previous hypotheses based on non-molecular evidence concerning the position of this genus within Boraginaceae tribe Boragineae. ITS-5.8S and trnL_UAA sequences from the nuclear and chloroplast non-coding genomes were obtained for four Cynoglottis taxa and selected members of the related genera Anchusa, Anchusella, Gastrocotyle, Brunnera and Pentaglottis. Cynoglottis is monophyletic, but neither trnL nor ITS support a close relationship with Brunnera, unlike previously supposed on morphological grounds. Brunnera is, instead, related to the southwestern European monotypic genus Pentaglottis, with which it forms a basal clade. ITS-5.8S sequences show that Anchusa thessala, a southeastern European annual species of Anchusa subg. Buglossellum, is sister to Cynoglottis and that the two taxa form a clade which also includes the Balkan endemic Gastrocotyle macedonica. Species of Anchusa subg. Anchusa form a separate lineage with high bootstrap support, suggesting that this heterogeneous genus is paraphyletic with respect to Cynoglottis. ITS sequences also discriminate between the Balkan-Apenninic diploid C. barrelieri and the Anatolian tetraploid C. chetikiana, albeit with low support. The molecular results are discussed in the light of karyological, morphological and chorological aspects.This work has been supported by M.I.U.R. 40% 2003 and the University of Firenze.