RAD sequencing,morphometry and synecology clarify the taxonomy of the Melica ciliata (Poaceae) complex in France and Poland

Melica (Poaceae) consist of about 92 species distributed across temperate regions of the world. Within section Dalycum, Melica ciliata sensu lato forms a taxonomic complex of several species and subspecies with clinal morphological variation causing conflicting identifications. To resolve taxonomic confusion, we used three complementary approaches, through molecular, morphological, and phytoecological analyses. The double-digest restriction-associated DNA markers significantly support the monophyly of three taxa: (i) the Mediterranean Melica magnolii, (ii) the Eurasian Melica transsilvanica subsp. transsilvanica, and (iii) the west-European M. ciliata subsp. glauca. This differentiation is corroborated by the analysis of 22 morphometric variables. Furthermore, phytoecological analysis of 221 floristic inventories revealed habitat distinctions among these taxa. Our approach of integrative taxonomy argues for a specific distinction for these three taxa, and we include a key to separate these forms. These new molecular data on the section Dalycum, subsection Ciliatae, call for further phylogenetic analyses including samples of M. ciliata subsp. ciliata and other East-Mediterranean and South African taxa.     

Soil Solution Chemistry Controlling the Field Distribution of Melica ciliata L.

Germination, establishment and growth of Melica ciliata L.,an 'acidifuge' species of rocky habitats in Europe, were studiedexperimentally and related to chemical properties of the soilsolution, including pH, base cation composition, Al concentrationand speciation, and Mn concentration. M. ciliata failed to establishin acid leptite soil. However, it was able to grow in solutionat pH 3·6 and exhibited vigorous growth at pH 3·9,a typical soil solution pH of leptite sites, which Melica isunable to colonize. Higher concentrations of Mn than those measuredin any leptite soil solutions did not influence growth. Exposureto 0·037 mmol l^-1 (1 mg l^-1) of Al³⁺, a concentrationusually exceeded in the soil solution of leptite sites, severelyretarded root growth. Speciation technique applied to Al insoil solutions obtained by centrifugation demonstrated a closerelationship between H⁺ and Al concentrations and, in particular,between H⁺ and free ionic (quickly reacting) Al species. Soilsolution concentrations of free ionic Al proved to be < 0·002mmol l^-1 in sites lacking Melica , but often > 0·10mmol l^-1 in site lacking Metalica . It is concluded that theinability of M. ciliata to establish in acid soils is not primarilydue to the high H⁺ concentration but to the high soil solutionconcentrations of Al, especially free Al ionic species at lowsoil pH.Copyright 1993, 1999 Academic Press Melica ciliata, distribution, soil solution, pH, aluminium speciation, manganese, base cations, iron     

Large-scale geographic patterns of genetic variation in Melica nutans, a widespread Eurasian woodland grass

 The geographic distribution of allozyme variation within the Eurasian boreo-nemoreal woodland grass Melica nutans L. has been investigated together with a minor subset of other Melica species. Twenty alleles were found at nine polymorphic loci in M. nutans. Allelic richness was highest in areas central in the species' European distribution, i.e. in southern Fennoscandia. High population densities, reducing the effects of genetic drift, as well as accumulation of variation through long-distance gene-flow from different marginal populations, is proposed to explain high allelic richness in this area. Several alleles showed geographic patterns in distribution and frequency variation. However, these patterns were not congruent, e.g. some alleles appear to have migrated to northern Europe from the south-west whereas others may have spread from the east. Genetic distances between geographic regions, each consisting of 2–6 populations, were generally low between all Fennoscandian, Russian and Siberian regions, but much higher between western and continental European regions. On the population level, cluster analysis grouped populations from Siberia, Russia, coastal and lowland areas in Fennoscandia and British Cumbria into one subcluster whereas other subclusters contained mainly south-west European populations or populations from almost throughout the distribution range. A scenario with several independent glacial refugia in central Europe, south-western Siberia and possibly western Norway, and subsequent colonisation of Fennoscandia mainly from the east, but with some long-distance gene-flow from central Europe, is proposed. Received April 3, 2002; accepted September 17, 2002 Published online: December 11, 2002     

Pollen morphology of the genus Iris L. (Iridaceae) from Croatia and surrounding area: taxonomic and phylogenetic implications

In Croatia and the surrounding area, the genus Iris is represented with about 20 Alpine-Dinaric, Mediterranean and Pannonian taxa from the subgenera Iris and Limniris. We researched pollen morphology of all taxa by using scanning electron microscopy. All pollen grains are sulcate with few palynological features, which could have taxonomic importance on different classification levels: shape of dry pollen grains; shape, outline and approximate size of hydrated pollen grains, approximate size of the sulcus, and ornamentation of the sulcus membrane and of the exine. At least four pollen types were recognised and taxonomically delimited to the series level: two characterise the subgenus Iris, section Iris (series Elatae and Pumilae), and two the subgenus Limniris, section Limniris (series Laevigatae, Sibiricae and Spuriae). Taxonomic implications of pollen morphology of their critical groups and taxa have been evaluated. Possible phylogenetic implications of pollen morphology of the genus Iris on the Southern European area were suggested: the subgenus Iris is more advanced than the subgenus Limniris; I. graminea seems to be the most primitive member of the subgenus Limniris on the Southern European territory; and pollen morphology of the population of the subspecies I. sibirica subsp. erirrhiza from the Mountain Bjelolasica could represent a link between the subgenera Limniris and Iris in the territory of Southern Europe. The possible pathway of evolution of the genus Iris on the territory of Southern Europe was suggested: from the subgenus Limniris, through the “linked taxon” I. sibirica subsp. erirrhiza from the Bjelolasica Mountain, to the subgenus Iris, series Pumilae, and finally to the series Elatae.     

HEARTWOOD FLAVONOIDS AND THE INFRAGENERIC RELATIONSHIPS OF RHUS (ANACARDIACEAE)

Heartwood flavonoids of 23 taxa of Rhus L. were surveyed in order to assess infrageneric relationships and classification. Fourteen flavonoids and two coumarins were detected in the heartwood extracts. All taxa were characterized by a flavonoid complement consisting of eight 5-deoxyflavonoids involving several aglycone classes (e.g., flavonols, flavones, aurones, chalcones and dihydroflavonols) and the aurone rengasin. None of the 5-hydroxyl analogs of the 5-deoxyflavonoids were detected in the heartwood extracts. Infraspecific flavonoid patterns were uniform in different populations, although the presence of 3′,4′-dihydroxyflavone 4′-O-β-glucoside varied in some taxa. Taxa of Rhus subgenus Rhus consistently differed from all taxa of Rhus subgenus Lobadium in lacking glycosides of fisetin, butein and 3′,4′-dihydroxyflavone. The major evolutionary trend in the heartwood flavonoids of Rhus appears to be accumulation of simple mono- or diglucosides. Data from heartwood flavonoids suggest that Rhus be treated as consisting of two subgenera (Rhus and Lobadium) and that subgenus Lobadium be divided into three sections.     

Micromorphological observations on leaf and pollen of Capparis L. sect. Capparis (Capparaceae)

Sect. Capparis is represented by a single species, Capparis spinosa L., divided into several intraspecific taxa showing plesiomorphic features and disjunct distributions in the Old World. Leaf surface and pollen features were investigated in the whole group by SEM and light microscope observations. The section is characterized by simple hairs, a reticulate to undulate cuticle, anomocytic stomata surrounded by a peristomal rim, and trizonocolporate, prolate pollen grains. The characteristics of the indumentum appear constant, while the studied taxa are fairly differentiated with respect to cuticular patterns and dimensions of the stomata, and show slight differences in pollen size and exine surface. This micromorphological evidence, coupled with other phenotypic features, supports the placement of this section at the base of the genus Capparis in the paleotropical area. Considering the striking geographic disjunction and symplesiomorphies of the group, its biogeographical and systematic aspects are also discussed.     

Silica bodies and hooked papillae in lemmas of Melica species (Gramineae: Pooideae)

Melica (sensu lato ) is a genus with approximately 80 species distributed in the temperate regions of both hemispheres. The genus has been better studied morphologically than anatomically, but morphological patterns have led to conflicting subdivisions either into sections, or by segregation of the genus Bromelica . The lemma epidermis is examined here under scanning electron microscopy (SEM) for 40 species of Melica to see if it exhibits characters of systematic significance. Silica bodies and hooked papillae are reported in the lemma for the first time in the genus. Patterns of variation are evident within four structures that terminate in points: prickles, hooks, hooked papillae and macro-hairs. Fourteen qualitative characters for lemma epidermis are used in phenetic analyses. The presence or absence of macro-hairs conflicts with some other patterns, but gives some support to the classification of Papp. Characters associated with the split of the clusters were micro-hairs (straight and geniculate) and the distribution and frequency of prickles in both costal and intercostal zones. As a conclusion, Melica is not a coherent taxonomic grouping and a simple segregation of Bromelica from Melica genus is rejected.  © 2003 The Linnean Society of London , Botanical Journal of the Linnean Society , 2003, 141, 447 – 463.     

A CHLOROPLAST-DNA PHYLOGENY OF THE WILD PERENNIAL RELATIVES OF SOYBEAN (GLYCINE SUBGENUS GLYCINE): CONGRUENCE WITH MORPHOLOGICAL AND CROSSING GROUPS

Hypotheses of evolutionary relationships among the Australian wild perennial relatives of soybean (Glycine subgenus Glycine) are based largely on patterns of meiotic pairing in intra- and interspecific experimental hybrids. This evidence has indicated a number of genome groupings within the subgenus but has not resolved most phylogenetic relationships. Restriction-endonuclease site variation of chloroplast DNA (cpDNA) within the perennial subgenus is reported here, representing a sampling of approximately 3% of the approximately 150-kilobase plastome. Seven hundred twenty-one unique restriction sites were compared within Glycine using 29 restriction endonucleases; 157 sites varied within the genus. Distance and parsimony methods using these data yielded congruent results, recognizing the existence of three major groups within subgenus Glycine: the species-rich and geographically diverse A clade consisting of G. canescens and related taxa; the B clade, which includes the stoloniferous species; and the C group, containing two species with distinctive curved pods. These results are in general agreement with hypotheses based on genome analysis; inconsistencies involve the inclusion of genetically divergent taxa such as G. falcata in well-supported plastome clades comprised of otherwise interfertile species. Such findings are not unexpected if crossing barriers are considered to be unique features of such anomalous species, paralleling their often numerous morphological and cpDNA autapomorphies. Consideration of cpDNA divergence within the three major clades of subgenus Glycine indicates that the rate of plastome evolution is uncoupled from rates of morphological or ecological diversification.     

Molecular phylogenetic analysis of Arenaria (Caryophyllaceae: tribe Arenarieae) and its allies inferred from nuclear DNA internal transcribed spacer and plastid DNA rps16 sequences

The systematics and phylogeny of the genus Arenaria and allied genera are unresolved. The use of morphological data has resulted in contradictory taxonomic concepts in the past due to their homoplastic nature. We present a phylogenetic analysis based on internal transcribed spacer (ITS) and rps16 sequence data of 140 (132 taxa) and 131 (120 taxa) accessions, respectively. Maximum parsimony and Bayesian analyses of each marker produced nearly congruent trees. Monophyly of Arenaria s.s. and Eremogone is confirmed here. Our results corroborate earlier results indicating that Arenaria subgenus Odontostemma is monophyletic, but outside the core group of Arenaria. Arenaria subgenus Solitaria is sister to Odontostemma and also not closely related to the latter; both of these subgenera are excluded from Arenaria and treated as distinct genera. The molecular data indicate that the ‘Arenaria s.s. clade’ consists of a few well‐supported subgroups and that the current subgeneric classification of the genus does not reflect evolutionary history. Arenaria subgenus Leiosperma is clearly monophyletic, but we reduce it to sectional level. Our molecular data show that the monotypic Arenaria subgenera Porphyrantha and Arenariastrum are nested in A. subgenus Arenaria, whereas subgenus Eremogoneastrum is included in Eremogone. None of the species‐rich sections in subgenus Arenaria is monophyletic. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 648–669.     

Chemosystematische Untersuchungen zur Phylogenese der Sammelgattung Lacerta (Reptilia: Sauria: Lacertidae)

Chemosystematic investigations concerning the phylogeny of the collective genus Lacerta (Reptilia: Sauria: Lacertidae) Investigated the albumins from 31 species and subspecies of the genus Lacerta s.1. by means of the immunoloical techniue Micro-Complement-fixation. Dendrograms were computed by means of the ktch-MargJiash algorithm. Moreover the taxa of the Lacerta saxicola and the Lacerta danfordi comlexes were analyzed by electrophoresis concerning 14 genetic loci. The results were discussel together with karyological and genitalmorphological data so far known yielding the following relationships: Lacerta graeca shows a closer relationshi to the subgenus Podarcis as does the quite closely related species air L. dugesii and L. perspicikata. Lacerta Iaevis probabl is connected closely to Podarcis, too. Lacerta parva shows a closer relationship to the “small izards” than to the sub-genus Lacerta s. str. A clear decision concerning the position of the Lacerta danfordi complex according to chemosystematical data was not possible. It is isolated from the other species probably about as long as L. puma. Biochemical differences between its “species” are too small to confirm their taxonomical revalorisation by Eiselt and Schmidtler Within the Lacerta saxicola complex the investigated taxa show a close relationship. Lacerta praticola and L. derjugini were definitly assigned to this group.     

A phenetic investigation of Vicia L.subgenus Vicia (Leguminosae, Vicieae)

Morphological data were gathered from 1539 herbarium specimens representing the taxa of Vicia subgenus Vicia. Specimens were scored for 43 vegetative, 84 inflorescence, 23 legume and 24 seed characters. The data were analysed using cluster analysis and ordination techniques. The results of the analysis were used to produce a classification of the subgenus. The proposed classification contains nine sections, nine series, 38 species, 14 subspecies and 22 varieties. The classification is discussed in relation to previous classifications.     

Some evidence for infrageneric classification in Ziziphora L. (Lamiaceae:Mentheae)

The genus Ziziphora L. (Lamiaceae) consists of annual herbs, except Ziziphora clinopodioides, and is represented by six taxa in the flora of Turkey: Z. clinopodioides, Z. capitata, Z. persica, Z. tenuior, Z. taurica subsp. taurica and Z. taurica subsp. cleonioides. They are strong aromatic herbs and important medicinal plants. This study has been conducted to determine the palynological and karyological features of these taxa. The pollen morphology and exine structure of Ziziphora species were investigated by light microscope and scanning electron microscope. Pollen grains were determined as hexacolpate, medium in size and a circular or elliptic amb. The shape of pollen grains varies from oblate to prolate spheroidal. Ornamentation types of pollen grains are bireticulate, microreticulate or perforate. Somatic chromosome number has been determined and counted as 2n = 16, 18. Results indicate that the pollen characters and chromosome numbers of the genus are valuable for taxonomic applications and may be useful for infrageneric classification.     

Giants and dwarfs: molecular phylogenies reveal multiple origins of annual spurges within Euphorbia subg. Esula

Euphorbia (Euphorbiaceae) comprises over 2150 species and is thus the second-largest genus of flowering plants. In Europe, it is represented by more than 100 species with highest diversity in the Mediterranean area; the majority of taxa belong to subgenus Esula Pers., including about 500 taxa. The few available phylogenetic studies yielded contrasting results regarding the monophyly of subg. Esula, and the phylogenetic relationships among its constituents remain poorly understood. We have sampled DNA sequences from the nuclear ribosomal internal transcribed spacer (ITS) and the plastid trnT-trnF region from about 100, predominantly European taxa of subg. Esula in order to infer its phylogenetic history. The plastid data support monophyly of subg. Esula whereas the ITS phylogeny, which is generally less resolved, is indecisive in this respect. Although some major clades have partly incongruent positions in the ITS and plastid phylogenies, the taxonomic content of the major terminal clades is congruent in both trees. As traditional sectional delimitations are largely not corroborated, an improved classification is proposed. Character state reconstruction illustrates that the annual life form developed independently several times in different clades of subgenus Esula from perennial ancestors, and that several morphological traits used in previous classifications of Euphorbia developed in parallel in different lineages.     

The role of chromosome changes in the diversification of Passiflora L. (Passifloraceae)

Passiflora L. has more than 575 species distributed especially in the Neotropics. The chromosome number variation in the genus is highly congruent with its main subgenera, but its basic chromosome number (x) and the underlying events responsible for this variation have remained controversial. Here, we provide a robust and well-resolved time-calibrated phylogeny that includes 102 taxa, and by means of phylogenetic comparative methods (PCM) we tested the relative importance of polyploidy and dysploidy events to Passiflora karyotype evolution and diversification. Passiflora arose 42.9 Mya, with subgenus diversification at the end of the Palaeogene (Eocene-Oligocene). The basic chromosome number of the genus is proposed as x?=?6, and a strong recent diversification found in the Passiflora subgenus (Miocene) correlated to genome size increase and a chromosome change from n?=?6 to n?=?9 by ascending dysploidy. Polyploidy, conversely, appeared restricted to few lineages, such as Astrophea and Deidamioides subgenera, and did not lead to diversification increases. Our findings suggest that ascending dysploidy provided a great advantage for generating long-term persistent lineages and promoting species diversification. Thus, chromosome numbers/genome size changes may have contributed to morphological/ecological traits that explain the pattern of diversification observed in the genus Passiflora.     

Genetic variation and biogeography of the disjunct Vitis subg. Vitis (Vitaceae)

Aim Vitis subg. Vitis provides an example of a plant disjunction occurring in the Northern Hemisphere. It shows broad morphological variation but is assumed to be a species complex with limited genetic differentiation. Based on a comprehensive sampling of taxa and polymorphism in both chloroplast and nuclear DNA, we assessed genetic variation within this subgenus. Our aims were to clarify the relationships among species and to examine their historical biogeography. Location Asia, Europe, North America. Methods We analysed a total of 30 species and putative hybrids from subgenus Vitis and examined the infra‐specific variation in some species. Polymorphism in chloroplast DNA was assessed in trnL and trnH–psbA–trnK sequences (c. 2170 bp) and in 15 microsatellite loci. We also obtained nuclear data for size variation at 24 microsatellite loci. Phylogenetic inference was performed with Bayesian analyses. A maximum parsimony network was constructed to depict the evolutionary relationships among haplotypes, and microsatellite data were also subjected to hierarchical clustering analysis using the Ward distance. In addition, we assessed size homoplasy by sequencing both chloroplast and nuclear microsatellite loci. Results Chloroplast polymorphisms resolved subgenus Vitis as a monophyletic group with limited genetic variation. The ancestral haplotypes were found in Eurasia. American taxa all harboured derived haplotypes. Most of them formed a monophyletic group that did not include Vitis californica. The four main haplotypes in Vitis vinifera corresponded to two different origins. Nuclear microsatellites indicated that genetic variation was especially large in North America. Asian species exhibited a lower level of nuclear divergence and the European V. vinifera corresponded to a differentiated nuclear lineage. Main conclusions We obtained some evidence that subgenus Vitis has an Asian origin and then dispersed to Europe and North America. Geographic separation was followed by diversification, presumably during the Pleistocene, resulting in phylogeographic patterns similar to other biota. In contrast to chloroplast DNA, nuclear DNA shows a larger than expected genetic variation. Our molecular data also highlight the need to re‐examine certain aspects of the current subgeneric classification.     

Chemotaxonomy and evolution of Plantago L.

In continuation of our investigations of the genus Plantago L. (Plantaginaceae), sixteen species were investigated with respect to water-soluble glycosides. The iridoids auroside, strictoloside and globularicisin, as well as poliumoside, 3-[(4--D-glucopyranosyloxy)phenyl]propionic acid and 2-[4-(-D-glucopyranosyloxy)phenyl]acetic acid were isolated from Plantago for the first time. The latter compound has not previously been isolated as a natural product. Sorbitol was the main carbohydrate in all the species investigated. The distribution of iridoids correlates well with the morphological classification of Rahn and also with a recently published molecular phylogenetic study of nuclear ribosomal and plastid DNA sequences. A new chemotaxonomic finding is an abundance of iridoid glucosides present in one of the two groups within subgenus Coronopus section Coronopus, while the other group lacks iridoids, supporting a subdivision of this section. Moreover, in subgenus Albicans, 10-benzoylcatalpol is a characteristic constituent of section Gnaphaloides, while the corresponding cinnamoyl ester globularin occurs in section Lanceifolia. In biosynthetic experiments, labelled epideoxyloganic acid and deoxygeniposidic acid were incorporated into aucubin and geniposidic acid in Plantago ovata, consistent with earlier findings, but no incorporation into asperuloside was observed. The evolution of biosynthetic pathways in Plantago is discussed.     

The biogeography of Gunnera L.: vicariance and dispersal

Aim The genus Gunnera is distributed in South America, Africa and the Australasian region, a few species reaching Hawaii and southern Mexico in the North. A cladogram was used to (1) discuss the biogeography of Gunnera and (2) subsequently compare this biogeographical pattern with the geological history of continents and the patterns reported for other Southern Hemisphere organisms. Location Africa, northern South America, southern South America, Tasmania, New Zealand, New Guinea/Malaya, Hawaii, North America, Antarctica. Methods A phylogenetic analysis of twenty‐six species of Gunnera combining morphological characters and new as well as published sequences of the ITS region, rbcL and the rps16 intron, was used to interpret the biogeographical patterns in Gunnera. Vicariance was applied in the first place and dispersal was only assumed as a second best explanation. Results The Uruguayan/Brazilian Gunnera herteri Osten (subgenus Ostenigunnera Mattfeld) is sister to the rest of the genus, followed sequentially upwards by the African G. perpensa L. (subgenus Gunnera), in turn sister to all other, American and Australasian, species. These are divided into two clades, one containing American/Hawaiian species, the other containing all Australasian species. Within the Australasian clade, G. macrophylla Blume (subgenus Pseudogunnera Schindler), occurring in New Guinea and Malaya, is sister to a clade including the species from New Zealand and Tasmania (subgenus Milligania Schindler). The southern South American subgenus Misandra Schindler is sister to a clade containing the remaining American, as well as the Hawaiian species (subgenus Panke Schindler). Within subgenus Panke, G. mexicana Brandegee, the only North American species in the genus, is sister to a clade wherein the Hawaiian species are basal to all south and central American taxa. Main conclusions According to the cladogram, South America appears in two places, suggesting an historical explanation for northern South America to be separate from southern South America. Following a well‐known biogeographical pattern of vicariance, Africa is the sister area to the combined southern South America/Australasian clade. Within the Australasian clade, New Zealand is more closely related to New Guinea/Malaya than to southern South America, a pattern found in other plant cladograms, contradictory to some of the patterns supported by animal clades and by the geological hypothesis, respectively. The position of the Tasmanian G. cordifolia, nested within the New Zealand clade indicates dispersal of this species to Tasmania. The position of G. mexicana, the only North American species, as sister to the remaining species of subgenus Panke together with the subsequent sister relation between Hawaii and southern South America, may reflect a North American origin of Panke and a recolonization of South America from the north. This is in agreement with the early North American fossil record of Gunnera and the apparent young age of the South American clade.     

Leaf epidermal studies in some Nigerian species of Ficus L. (Moraceae)

Forty-two leaf epidermal features in 20 Nigerian species of Ficus, representing three of the four subgenera and four of the seven sections of the genus found in Africa have been examined. Discontinuities in these characters clearly separated the three subgenera studied, namely, Ficus, Sycomorus and Urostigma, and supported the earlier sectional classification of the subgenus Urostigma (i.e. sections Sycidium, Galoglychia and Urostigma). The subsectional boundaries among the members of the section Galoglychia studied, however, had little backing from their leaf epidermal characteristics due to extensive overlap of the features. The results of the present studies support the genetic basis of the qualitative and quantitative variations in the leaf epidermal cells, stomata and trichomes in the Nigerian Ficus and the present and potential applications of these features in diagnostic and taxonomic investigations in the genus are discussed.