The phylogenetic placement of Ostropales within Lecanoromycetes (Ascomycota) revisited

Results of molecular studies regarding the phylogenetic placement of the order Ostropales and related taxa within Lecanoromycetes were thus far inconclusive. Some analyses placed the order as sister to the rest of Lecanoromycetes, while others inferred a position nested within Lecanoromycetes. We assembled a data set of 101 species including sequences from nuLSU rDNA, mtSSU rDNA, and the nuclear protein-coding RPB1 for each species to examine the cause of incongruencies in previously published phylogenies. MP, minimum evolution, and Bayesian analyses were performed using the combined three-region data set and the single-gene data sets. The position of Ostropales nested in Lecanoromycetes is confirmed in all single-gene and concatenated analyses, and a placement as sister to the rest of Lecanoromycetes is significantly rejected using two independent methods of alternative topology testing. Acarosporales and related taxa (Acarosporaceae group) are basal in Lecanoromycetes. However, if the these basal taxa are excluded from the analyses, Ostropales appear to be sister to the rest of Lecanoromycetes, suggesting different ingroup rooting as the cause for deviating topologies in previously published phylogenies.     

Does Lilium bosniacum merit species rank? A classical and molecular-cytogenetic analysis

Four populations of the Balkan endemic taxon Lilium bosniacum from Bosnia and Herzegovina were investigated. Conventional karyological study did not reveal any important differences. Molecular-cytogenetic studies pointed out certain interpopulation and intrapopulation variability, and important differences in organization of ribosomal genes in regard to its closest relative L. carniolicum. The results of fluorochrome banding and FISH experiment for L. bosniacum were reported here for the first time. Differences occurred in the number and position of 18S-5.8S-26S ribosomal gene loci for some individuals from population Kladanj. Heterochromatic bands revealed with DAPI after FISH experiment were constant. All investigated populations possess the same number of active NORs except some individuals from Kladanj population. Genome size and GC-bases percentage, estimated by flow cytometry, did not show any significant difference among the populations. However, the present results reveal clear interspecific differentiation between two endemics, L. carniolicum and L. bosniacum.     

Relationships between five species ofLolium (Poaceae)

Morphological analysis and electrophoresis of seed proteins of fiveLolium species disclosed that they form two distinct groups corresponding to those recognized from compatibility data.Lolium temulentum andL. remotum of the self-pollinated group were shown to be distinct but closely related species. Morphological intergradation and high similarities between protein profiles ofL. perenne, L. multiflorum andL. rigidum (cross-pollinated species) suggest little genetic differentiation between these taxa. This implies that treatment at the infraspecific level might better accomodate the data.     

Freshwater gomphonemoid diatom phylogeny: preliminary results

A cladistic analysis of eleven freshwater gomphonemoid diatom taxa yielded three equally most parsimonious cladograms. The three cladograms suggest Gomphoneis Cleve is non-monophyletic as originally and recently circumscribed. Gomphoneis elegans and G. transsilvanica (Pant.) Krammer are more closely related to Gomphopleura Reichelt ex Tempère than to other members of the genus. Gomphoneis geitleri Kociolek & Stoermer appears to represent a monotypic lineage. Among the taxa with differentiated apical pore fields Gomphonema kaznakowi Mereschkowsky appears to be primitively astigmate.The three cladograms vary with regard to systematic affinities among the Gomphoneis herculeana (Ehrenberg) Cleve group, Miocene gomphonemoid species from Idaho (USA), gomphonemoid taxa from East Africa and Gomphonema sensu stricto. Despite variability in number of puncta rows per stria and presence of an axial plate in the Miocene species from North America, inclusion of these taxa in the analysis provided better resolution of relationships than if they were excluded.     

Implications of molecular systematic analyses on the conservation of rare and threatened taxa: Contrasting examples from Malvaceae

Systematic research provides essential evidence for setting conservation priorities for rare and endangered taxa. Phylogenetic analyses can identify cryptic, genetically distinct lineages as well as actively interbreeding, and hence, non-distinctive lineages earlier perceived as separate taxa. A major aim of this study was to identify genetically distinct, rare lineages within two Malvaceae sister-genera, Sidalcea and Eremalche. The focus was two taxon-pairs each consisting of one rare and one more common taxon. The results demonstrate that even within two closely related genera, with a large number of rare taxa, molecular phylogenetic analyses can reveal contrasting degrees of evolutionary divergence and thus contrasting conservation implications for threatened taxa. Contrary to expectations, the substitution rate in the nuclear ribosomal transcribed spacers for annual Eremalche did not correspond to the faster evolutionary rate of annuals – compared to perennials – detected earlier within Sidalcea. Branch lengths in the (annual) Eremalche clade were shorter than those of annual members of Sidalcea. The phylogenetic analyses showed that the rare and endangered S. keckii and E. kernensis each are most closely related to a common species that has been regarded as insufficiently distinct to warrant separate taxonomic status. An additional aim of the study was to test the utility of the Phylogenetic Diversity (PD) measure to formalize the procedure of prioritizing conservation efforts. The measure demonstrated S. keckii (but not E. kernensis) to be genetically distinct from its closest relative and a good candidate for conservation. The PD measure was earlier used for assessing conservation priorities for areas, but proved useful to more objectively suggest conservation priorities among threatened taxa. Because this measure is calculated directly from the data, it retains more character information and gives a better representation of genetic diversity than other measures relying on tree topologies.     

Molecular phylogeny of Phebalium (Rutaceae: Boronieae) and related genera based on the nrDNA regions ITS 1+2

Parsimony analyses of the internal transcribed spacer regions of nuclear ribosomal DNA (ITS 1 & ITS 2) for 38 taxa sampled from the Phebalium group (Rutaceae: Boronieae) and two outgroups confirm that, with the exception of Phebalium sensu stricto and Rhadinothamnus, six of the currently recognised genera within the group are monophyletic. The data indicate that Phebaliums. str. is paraphyletic with respect to Microcybe, and Rhadinothamnus is paraphyletic with respect to Chorilaena. Rhadinothamnus and Chorilaena together are the sister group to Nematolepis. Drummondita, included as an outgroup taxon, clustered within the ingroup as sister to Muiriantha and related to Asterolasia.The phylogeny suggests that the evolution of major clades within a number of these genera (e.g. Phebalium) relates to vicariance events between eastern and south-western Australia. Leionema is an eastern genus, with the most basal taxon being the morphologically distinct Leionema ellipticum from northern Queensland. Leionema also includes one species from New Zealand, but this species (as with some others) proved difficult to sequence and its phylogenetic position remains unknown. Taxonomic changes at the generic level are recommended.The authors wish to thank Paul G.Wilson, PERTH, for advice and discussion, and Paul Forster, BRI, for collecting and providing material of Leionema ellipticum. The project was supported by a Melbourne University Postgraduate Award (to BM), the Australian Biological Resources Study (ABRS), Australian Systematic Botany Society and Wolf Den (Australia) Investments.     

Mikromorphologie der Samenoberflächen der GattungEuphorbia

Seed coat surfaces of 127 species, representing 23 sections of the genusEuphorbia L. (Euphorbiaceae) have been examined with the scanning electron microscope (SEM). Various surface patterns and cell wall structures are described. In some groups the seed coat is rather uniform (e.g. in sect.Euphorbium), in other sections even closely related species can be separated by seed coat morphology (e.g.Tithymalus). The taxonomic application of testa micromorphology and the possible systematic position of certain taxa are briefly discussed. The seed coat cells of all species in sect.Tithymalus show remarkable intercellulares filled with small particles, which are described in detail for the first time.

     

rbcL sequences support exclusion ofRetzia,Desfontainia, andNicodemia from theGentianales

The taxonomic positions ofRetzia, Desfontainia, andNicodemia have been much discussed, and all three genera have been included inLoganiaceae (Gentianales). We have made a cladistic analysis ofrbcL gene sequences to determine the relationships of these taxa toGentianales. Four newrbcL sequences are presented; i.e., ofRetzia, Desfontainia, Diervilla (Caprifoliaceae), andEuthystachys (Stilbaceae). Our results show thatRetzia, Desfontainia, andNicodemia are not closely related toLoganiaceae or theGentianales. Retzia is most closely related toEuthystachys and is better included inStilbaceae. The positions ofDesfontainia andNicodemia are not settled, butDesfontainia shows affinity for theDipsacales s.l. andNicodemia for theLamiales s.l.     

Phylogenetic relationships in theCampanulales based onrbcL sequences

Phylogenetic relationships within the angiosperm orderCampanulales were investigated by comparative sequencing of the chloroplast generbcL. CompleterbcL sequences were obtained for ten species in six families within the order. These data were analyzed along with previously publishedrbcL sequences from other taxa (for a total of 117 species) within the subclassAsteridae and outgroups, producing 32 equally parsimonious trees. A subset consisting of 44 of these taxa was then chosen and more rigorous analyses performed, resulting in four equally parsimonious trees. Results indicate that two major clades roughly corresponding to traditionally circumscribedAsterales andCampanulales exist as sister taxa. In particular, therbcL trees indicate thatSphenoclea is not a member ofCampanulales orAsterales, thatPentaphragma is more closely allied toAsterales thanCampanulales, that theCyphiaceae are not monophyletic, thatCampanulaceae andLobeliaceae are not sister taxa, and thatStylidiaceae are correctly placed withinCampanulales.     

Phylogenetic analysis of Leucojum and Galanthus (Amaryllidaceae) based on plastid matK and nuclear ribosomal spacer (ITS) DNA sequences and morphology

Phylogenetic analyses of the monocotyledonous genera Leucojum and Galanthus (Amaryllidaceae, Asparagales), using plastid (trnL-F and matK) and largely non-coding nuclear ribosomal (ITS) DNA sequences show the two to be closely related to Lapiedra, Narcissus, Vagaria, Pancratium and Sternbergia. We compare the results obtained with a combined parsimony analysis of these nucleotide sequences with that of a matrix of morphological characters. The sampling included all species of Leucojum and most species of Galanthus (representing all series and subseries of the genus) and used as outgroup the above mentioned genera of Amaryllidaceae shown to be close relatives. The plastid, nuclear and morphological data were analysed independently and in combination, showing that the boundaries between the two genera are not appropriate. Galanthus is monophyletic but embedded in Leucojum. On the basis of chromosome numbers and floral characters Leucojum has been previously divided into four subgenera, which have been accepted as genera by some authors. In our phylogenetic analyses (separate as well as combined), Leucojum species are separated in two primary clades corresponding to L. subgenera Ruminia + Acis and L. Leucojum + Aerosperma. The taxonomic implications of this pattern are discussed, and an alternative classification is proposed. Finally, biogeographic relationships of species of both Leucojum and Galanthus are discussed, emphasising the possible origin of the narrowly distributed taxa of Leucojum relative to the widespread species.     

Phylogeography of Indonesian and Sino-Himalayan region bush warblers (Cettia, Aves)

We present a hypothesis for the phylogeny and phylogeography of a group of bush warblers in the genus Cettia, based on parts of the mitochondrial Cytochrome b gene and the nuclear myoglobin intron II (in all 1.7 kb). Ancestral areas were reconstructed by dispersal-vicariance analysis and constrained Bayesian inference. The results suggest that the insular taxa in the Cettia vulcania group are most closely related to Cettia flavolivacea, and originated from a dispersal by an ancestral population in the Himalayas towards the south, to the Sunda region. From this population, a second dispersal along a different route colonized China and northern Vietnam. Hence, the Chinese taxon intricata and Vietnamese oblita, currently allocated to C. flavolivacea, are more closely related to the vulcania group than to the other taxa in the flavolivacea group, and we propose that they be treated as conspecific with C. vulcania, restricting C. flavolivacea to Myanmar and the Himalayas.     

Cytotaxonomische Untersuchungen in der Artengruppe desRanunculus alpestris (Ranunculaceae)

Morphological and cytological investigations as well as crossing experiments were carried out with the 5 species of theRanunculus alpestris group (R. alpestris L.,R. traunfellneri Hoppe,R. bilobus Bertol.,R. crenatus Waldst. etKit.,R. magellensis Ten.). A key to the species is presented; localities and distribution are given in addition to extensive diagnoses. Crossing experiments between the 5 taxa were successful (F₁-F₃ individuals, backcross types, tripeland quadrupelbastards); the morphology of the experimentally obtained F₁-hybrids was mostly intermediate. All 5 species as well as all hybrids have a chromosome number of 2n = 16; there is no statistically significant difference between the karyotypes of the 5 taxa. According to the results of the morphological investigations and the crossing experiments we can distinguish 2 subunits of very closely related species: a)R. alpestris, R. traunfellneri, R. bilobus; b)R. crenatus, R. magellensis. The speciation within the group ofRanunculus alpestris is discussed.

     

Genome relationships among Lotus species based on random amplified polymorphic DNA (RAPD)

The ability of random amplified polymorphic DNA (RAPD) to distinguish among different taxa of Lotus was evaluated for several geographically dispersed accessions of four diploid Lotus species, L. tennis Waldst. et Kit, L. alpinus Schleich., L. japonicus (Regel) Larsen, and L. uliginosus Schkuhr and for the tetraploid L. corniculatus L., in order to ascertain whether RAPD data could offer additional evidence concerning the origin of the tetraploid L. corniculatus. Clear bands and several polymorphisms were obtained for 20 primers used for each species/accession. The evolutionary pathways among the species/accessions presented in a cladogram were expressed in terms of treelengths giving the most parsimonious reconstructions. Accessions within the same species grouped closely together. It is considered that L. uliginosus which is most distantly related to L. corniculatus, may be excluded as a direct progenitor of L. corniculatus, confirming previous results from isoenzyme studies. Lotus alpinus is grouped with accessions of L. corniculatus, which differs from previous studies. With this exception, these findings are in agreement with previous experimental studies in the L. corniculatus group. The value of the RAPD data to theories on the origin of L. corniculatus is discussed.     

Nutlet sculpturing inScutellaria sect.Resinosa (Lamiaceae) and its taxonomic utility

Nutlet sculpturing of 22 taxa ofScutellaria sect.Resinosa, representing 18 of its 19 species, and the monotypic genusSalazaria was studied by scanning electron microscopy (SEM). The nutlets of sect.Resinosa, while remarkably variable among themselves, differed from hypothetically closely related sections in lacking circumferential banding and sessile glands. Species could be grouped by surface sculpturing into seemingly meaningful phenetic clusters.Scutellaria potosina var.platyphylla was sufficiently distinct micromorphically so as to be elevated to specific status asScutellaria platyphylla comb. and stat. nov. The significance of nutlet microcharacters within sect.Resinosa is discussed, especially as compared to closely related sections. The genusSalazaria had nutlet sculpturing quite different from all taxa belonging to sect.Resinosa.     

Molecular Phylogenetic Relationships Among Crested-tailed Mice (Genus <Emphasis Type="Italic">Habromys</Emphasis>)

We examined genealogical relationships among six of seven species of crested-tailed mice (Habromys chinanteco, H. delicatulus, H. ixtlani, H. lepturus, H. lophurus, and H. simulatus) using DNA sequence data from the cytochrome-b gene. Gene trees based on maximum likelihood, Bayesian inference and maximum parsimony were largely congruent in that H. lepturus and H. ixtlani were closely related and formed the sister group to H. lophurus. All analyses also arranged H. chinanteco and H. simulatus as sister taxa. These results are concordant with the phenetic groupings of Carleton et al. (2002) based on morphology. Our unweighted maximum parsimony trees did not resolve placement of H. delicatulus relative to other taxa. However, analyses using weighted maximum parsimony, maximum likelihood and Bayesian inference optimality criteria recovered a sister group relationship between H. delicatulus and the clade comprised of ((H. lepturus H. ixtlani)(H. lophurus)). This relationship differs from the overall phenetic similarity of H. delicatulus with H. simulatus and H. chinanteco, influenced by the small size of these three taxa, but is consistent with some derived features of the phallus (Carleton et al., 2002). Based on our sequence data, a specimen from Michoacán, México, recently assigned to Peromyscus sagax likely was inadvertently misidentified (Tiemann-Boege et al., 2000) and actually represents a new locality record for H. delicatulus. Finally, we comment on the conservation status of species of Habromys.     

Phylogenetic relationships in theSideritis leucantha group (Lamiaceae)

Six closely related taxa of the sect.Eusideritis of the genusSideritis (S. leucantha, S. pusilla, S. flavovirens, S. granatensis, S. biflora andS. osteoxylla) are analysed to elucidate their phylogenetic relationships and position within the sect.Eusideritis. Meiotic behaviour, karyotype features, size and fertility of pollen grains, DNA amounts and seed protein profiles are reviewed. A polyploid origin of the group (from x = 7) and the further diversification through dysploidy and chromosome repatterning is postulated.S. osteoxylla is apparently of hybrid origin.     

Species delimitation in the Luziola peruviana (Poaceae) complex

Luziola is a small genus from aquatic environments of the New World. The most widespread species in the genus isL. peruviana. Morphological variation has been documented for this species throughout its entire range of distribution, however this variation has been difficult to characterize. A population aggregation analysis was performed in order to determine how many species can be identified when analyzing characters from the individuals of this taxon as well as with individuals from closely related taxa such asL. divergens, L. doelliana, andL. pittieri. Multivariate analysis, ANOVA and ANCOVA analyses were also performed to detect if quantitative morphological variation is related to altitude and latitude. Results indicate that within this complex only one species should be recognized, referable toL. peruviana, and that plants near the equator are larger, and have larger lemmas and paleas in both female and male flowers. An identification key for the nine species now recognized inLuziola is included.     

The phylogenetic positions of the conifer genera Amentotaxus,Phyllocladus, and Nageia inferred from 18s rRNA sequences

To determine the evolutionary positions of the conifer genera Amentotaxus, Phyllocladus, and Nageia, we obtained 18S rRNA sequences from 11 new taxa representing the major living orders and families of gymnosperms. With the published Chlamydomonas as an outgroup, phylogenetic analyses of our new data and available sequences indicate that (1) the Gnetales form a monophyletic group, which is an outgroup to the conifers, (2) the conifers are monophyletic, (3) Taxaceae, Cephalotaxaceae, Cupressaceae, and Taxodiaceae form a monophyletic group, (4) Amentotaxus is closer to Torreya than to Cephalotaxus, suggesting that Amentotaxus is better to be classified as a member of Taxaceae, (5) Phyllocladus, Dacrycarpus, Podocarpus, and Nageia form a monophyletic group, and (6) Pinaceae is an outgroup to the other families of conifers. Our finding that Phyllocladus is a sister group of the Podocarpaceae disagrees with the suggestion that the phylloclade of the genus is an ancient structure and that the genus is a terminal taxon within the Podocarpaceae. The genus Nageia is more closely related to Podocarpus than to Dacrycarpus and was derived from within the Podocarpaceae. In conclusion, our data indicate that in conifers, the uniovulate cone occurred independently in Taxacaeae and Cephalotaxaceae, and in Podocarpaceae after the three families separated from Pinaceae, and support the hypothesis that the uniovulate cone is derived from reduction of a multiovulate cone.Correspondence to: S.-M. Chaw     

Pollen morphology of Stachys (Lamiaceae) in Iran and its systematic implication

Pollen grains of 30 taxa of the genus Stachys (29 spp. and one subsp.), representing 9 of the currently recognized sections and 1 species of the closely related genus Sideritis (Si. montana) distributed in Iran were examined by light and scanning electron microscopy. Twenty-eight taxa are studied for the first time under aspects of pollen morphology. The basic shape of the pollen grains in most taxa studied is prolate-spheroidal, but subprolate, spheroidal and oblate-spheroidal pollen grains can also be found in few species. The grains are usually tricolpate (the amb triangular), but also tetracolpate (the amb circular to more or less square) in some species (S. iberica, S. atherocalyx and Si. montana). The surface is microreticulate (the frequent type), reticulate, perforate, foveolate-psilate or foveolate. The lumina are separated by smooth or sinuate muri which make them polygonal, more or less rounded and elongate. Major pollen morphological features of the taxa studied are compared and discussed on the basis of taxonomical concepts. In some cases, these characters are useful in delimitation of formerly introduced sections while they mostly provide further characters in separating related species from each other. For example, all members of S. sect. Aucheriana are characterized by elongated lumina. Based on the oblate-spheroidal shape of its pollen as well as tetracolpate aperture type, the results of the present study confirm sect. Pontostachys as including S. angustifolia, S. iberica, S. sparsipilosa as well as S. atherocalyx. Our results also suggest that although some species like S. fruticolosa and S. lavandulifolia are morphologically well characterized, they cannot be separated from other species of Stachys based on pollen morphology.     

Phylogeny of Oedogoniales (Chlorophyceae, Chlorophyta) inferred from 18S rDNA sequences with emphasis on the relationships in the genus Oedogonium based on ITS-2 sequences

The phylogeny of Oedogoniales was investigated by using nuclear 18S rDNA sequences. Results showed that the genus Oedocladium, as a separated clade, was clustered within the clade of Oedogonium; whereas the genus Bulbochaete was in a comparatively divergent position to the other two genera. The relationship among the species of Oedogonium was discussed, focusing on ITS-2 phylogeny analyzed combining with some morphological characteristics. Our results showed that all the dioecious nannandrous taxa involved in this study were resolved into one clade, while all the monocious taxa were clustered into another clade as a sister group to the former. The report also suggests that the dioecious macrandrous taxa form a paraphyly and could be more basally situated than the dioecious nannandrous and the monoecious taxa by means of molecular phylogeny and morphotype investigations.