Crossability analysis in subgenus Tulipa of the genus Tulipa L.

A range of almost 1400 cross combinations has been carried out between 31 species of Tulipa subgenus Tulipa. Most of the sections are well separated reproductively from each other. The designated species aggregates in section Clusianae appeared to be intersterile. Also in section Kolpakowskianae the species, as far as used, cannot be combined. Some crosses between species of section Tulipanum succeeded. The species of sections Eichleres and Tulipa (including the cultivated tulip T gesneriana) could be combined in various ways, although the highest level of seed set was achieved within either section. The crossability data support the current taxonomic division. The level of reproductive isolation between sympatric species is discussed, and the importance of crossability studies for plant breeding in Tulipa pointed out briefly.     

Molecular phylogeny of the genus Helix (Pulmonata: Helicidae)

The vast diversity of land snail forms is insufficiently understood even in seemingly familiar taxa. This holds for Helix Linnaeus, 1758, a genus with several common edible species which comprises the largest Western Palearctic snails. The taxonomy of this genus, which has a centre of diversity in the eastern Mediterranean, has recently undergone significant changes, in both the delimitation of the genus itself and its species‐level systematics. Here, we compare the lineage diversity of Helix, as revealed by two mitochondrial markers, with the conclusions of the recently published morphology‐based taxonomic revision. For the molecular analysis, we assembled a representative data set covering almost all species of the genus as recognized by the mentioned revision. We obtained sequences not only from fresh and preserved soft tissues but also from dried tissue remains (some of them decades old) from shell collections. Our results show that the genus Helix, in the narrow sense proposed by recent studies, is paraphyletic because the genus Tacheopsis was unambiguously revealed as one of the tip branches of Helix. The monophyly of several species, as presently recognized, was not supported; partly, this may be attributed to a lineage diversity overlooked so far. This holds also for the type species of the genus, H. pomatia, which comprises at least one additional lineage. Greece, the Aegean and western Turkey is the core area for the diversity of Helix and its relatives, and the region is probably a major long‐term refuge for large Helicidae. The highest species diversity is found along the Alpide belt from the western Balkans to southern Turkey. The diversity of Helix in Europe, north of Greece and the Apennines, is a result of a single European radiation. Our data also suggest that past human activities likely influenced the present‐day distributions of some species.     

Phylogenetic systematics of Erythronium (Liliaceae): morphological and molecular analyses

For a nearly complete set of species of Erythronium (Liliaceae), we examined two plastid loci (the rps16 intron and the 5′ trnK intron, excluding the matK exon), one nuclear locus (nrITS) and morphology to evaluate species relationships and that of Erythronium to Amana, the putatively most closely allied genus. A matrix of morphological characters was developed through observation of around 900 living and herbarium specimens; evolution of these was examined using character optimization on the combined (total‐evidence) tree. Parsimony methods were used to examine the morphological and molecular data sets produced, both separately and in combination, with Bayesian methods also used on the molecular data sets. These established that the genus is probably sister to Amana (although most analyses placed Amana inside Erythronium) and that Tulipa is sister to the pair of Erythronium and Amana. Within Erythronium, there are three strongly supported geographically distinct clades: (1) Eurasian and (2) eastern and (3) western North American. Separation of species in these three clades is less clear, particularly among the western North American taxa. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, ●●, ●●–●●.     

Genetic diversity among African great apes based on mitochondrial DNA sequences

The mitochondrial DNA D-Loop region was sequenced, analyzed and used as a molecular marker for populations of chimpanzee (Pan troglodytes), bonobo (P. paniscus) and gorilla (Gorilla gorilla and G. beringei), and also compared with data previously reported for these taxa. In Gorilla, a main disjunction between western (G. gorilla) and eastern (G. beringei, including G. b. graueri) species was observed, as well as high mitochondrial diversity within the western species. The genetic distance values within G. gorilla (0.14) were higher than those between subspecies (eastern lowland and mountain 0.12). Likewise, values of genetic diversity within this species (0.05) were higher than those between species (western and eastern lowland gorilla 0.04). Similarly, in genus Pan a main differentiation between western (P. t. verus) and central forms (P. t. troglodytes and P. t. schweinfurthii) was observed. The obtained values of genetic distance and genetic diversity revealed that the central subspecies are closer to each other than either of them is to the western one, while bonobos composed a distinct clade that expresses a well-defined specific identity. The current distribution, phylogeny and levels of genetic diversity in African great ape populations are consistent with the hypothesis that Pleistocene climatic events led to cyclical periods of isolation in forest refugia followed by expansion and dispersal. The implications of this high level of genetic diversity for taxonomic classification, wildlife management and conservation are discussed.     

Evolutionary and biogeographical patterns within the smelt genus Hypomesus in the North Pacific Ocean

Aim We address questions about trans‐Pacific distributions of marine organisms and the North Pacific Ocean as a centre of marine biodiversity through a phylogenetic and biogeographical study of a pan‐Pacific genus of Northern Hemisphere smelts (Hypomesus, Pisces: Osmeridae). Location North Pacific Ocean. Methods Relationships of the five species of Hypomesus from throughout the North Pacific were reconstructed through maximum likelihood and Bayesian phylogenetic analyses of sequence data from two mitochondrial (cytb, 16S) and three nuclear (ITS2, S71, RAG1) gene regions of five to 25 individuals per species, totalling 3588 characters. The resulting phylogenies were used to test hypotheses of species relationships and geographical origins using both dispersal‐based and maximum likelihood methods for inferring ancestral areas (lagrange ). Cytb sequence divergence and a Bayesian approach (beast ) were used to estimate the timeframe of Hypomesus evolution, which was compared with work on similarly distributed taxa. Results Hypothesized trans‐Pacific Ocean relationships based on lateral line scale counts were not supported by the phylogeny, suggesting parallel evolution of this phenotype, although we found one such relationship between the western H. japonicus and the two eastern Pacific species (H. pretiosus and H. transpacificus). Dispersalist approaches rejected an early proposal of a double‐compression vicariant mechanism as well as an eastern Pacific origin. Results from the lagrange analysis suggested a more widespread ancestor, although also supporting a role for the western Pacific. Divergence estimates suggested that most splits between species occurred in the mid‐Miocene, and the most recent speciation event, between the eastern Pacific species, occurred in the Pliocene to early Pleistocene. Main conclusions Our molecular data indicate that the character historically used to define relationships within Hypomesus, lateral line scale count, does not reflect ancestry within the genus. Biogeographical reconstructions suggest an important role for the western North Pacific in the diversification of Hypomesus. While uncertainty remains over the date of origin for this genus, estimates place the divergences during periods of climatic cooling that have been important in generating diversity in a number of similarly distributed organisms. Additional comparative data will provide further insight into the relative importance of the western region in generating diversity in the North Pacific Ocean.     

Patterns of freshwater biodiversity in Europe: lessons from the spring snail genus Bythinella

Aim We analyse patterns of biodiversity in the spring snail genus Bythinella, a group of highly isolated and stenotopic freshwater species. We aim to test: (1) whether there are European areas of increased diversity (i.e. ‘hotspots’), (2) whether the potential hotspots inferred show qualitative differences in biodiversity characteristics such as endemicity, distinctiveness of taxa, age of lineages or degree of fragmentation, and (3) whether these hotspots match the Pleistocene refugia of Bythinella spp. Location Europe, Asia Minor. Methods The analyses are based on genetic data from 717 Bythinella specimens sampled at 194 sites. We used haplotypes as operational units in all analyses. To test hypothesis 1, mean pairwise genetic distances between Bythinella populations within each 1° × 1° geographical grid cell sampled in Europe were calculated. Within individual mountain ranges, grid cells with high diversity were grouped with neighbouring ones and hotspots were identified based on pre‐defined criteria. Then, to test hypothesis 2, different biodiversity indices of these regions were calculated and compared. Finally, to test hypothesis 3, the spatial distribution of the identified hotspots was compared with the known Pleistocene refugia of Bythinella spp. Results Five areas showed increased levels of genetic diversity: the Massif Central/Pyrenees, the western and eastern Alps, and the western and eastern Carpathians. These regions showed qualitative differences in biodiversity, with the eastern Carpathians holding the highest number of (endemic) haplotypes, the oldest and most distinct lineages and the highest degree of fragmentation. Only three of the five detected hotspots matched previously identified Pleistocene refugia for Bythinella spp. Main conclusions The genetic diversity of Bythinella spp. is not randomly distributed throughout Europe. Some of the hotspots we identify coincide with those found in other freshwater taxa; others have not previously been reported. Thus, spring organisms may reflect a unique evolutionary history that is distinct from lentic and lotic taxa. Our findings may be useful for conservation purposes even though the species‐level taxonomy of the genus is still under discussion.     

Phylogenetic relationships in the genus Leontopodium (Asteraceae: Gnaphalieae) based on AFLP data

The genus Leontopodium comprises 30–41 species. The centre of diversity is the Sino‐Himalayan region in south‐western China, where about 15 species occur. The two species native to Europe, L. alpinum (known as the common ‘Edelweiss’) and L. nivale, are part of the cultural heritage of the people living there. Despite its importance, very little is known about the systematics of the genus. Because recent molecular studies have shown that species within this genus are closely related and difficult to distinguish with rDNA and cpDNA data, we used AFLPs to obtain a more detailed understanding of the phylogeny of the genus. Our main aims were as follows: (1) to clarify species relationships within the genus; and (2) to reveal information about the biogeography of the genus. We used AFLPs with six primer combinations to investigate 216 individuals in 38 populations of 16 different species. With AFLPs, we were able to recognize 10 different groups, all of which had strong bootstrap support. These results were also congruent with the morphology‐based taxonomy of the genus. Most private and rare fragments were found in the Yunnan region (south‐western China) relative to Europe and Mongolia/central China, suggesting a long‐lasting in situ history of populations in the centre of diversity of the genus. Our results illustrate the utility of AFLPs to resolve phylogenetic relationships between these closely related species. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 165 , 364–377.     

Taxonomic surrogacy in biodiversity assessments,and the meaning of Linnaean ranks

The majority of biodiversity assessments use species as the base unit. Recently, a series of studies have suggested replacing numbers of species with higher ranked taxa (genera, families, etc.); a method known as taxonomic surrogacy that has an important potential to save time and resources in assesments of biological diversity. We examine the relationships between taxa and ranks, and suggest that species/higher taxon exchanges are founded on misconceptions about the properties of Linnaean classification. Rank allocations in current classifications constitute a heterogeneous mixture of various historical and contemporary views. Even if all taxa were monophyletic, those referred to the same rank would simply denote separate clades without further equivalence. We conclude that they are no more comparable than any other, non‐nested taxa, such as, for example, the genus Rattus and the phylum Arthropoda, and that taxonomic surrogacy lacks justification. These problems are also illustrated with data of polychaetous annelid worms from a broad‐scale study of benthic biodiversity and species distributions in the Irish Sea. A recent consensus phylogeny for polychaetes is used to provide three different family‐level classifications of polychaetes. We use families as a surrogate for species, and present Shannon‐Wiener diversity indices for the different sites and the three different classifications, showing how the diversity measures rely on subjective rank allocations.     

A review of cross‐backed grasshoppers of the genus Dociostaurus Fieber (Orthoptera: Acrididae) from the western Mediterranean: insights from phylogenetic analyses and DNA‐based species delimitation

Phylogenetic analyses and species delimitation methods are powerful tools for understanding patterns of species diversity. Given the current biodiversity crisis, such approaches are invaluable for urgent assessment and delimitation of truthful species, particularly of endangered and morphologically cryptic taxa from vulnerable areas submitted to strong climate change and progressive human intervention such as the M editerranean region. In this study, we applied two DNA ‐based species delimitation methods and performed a B ayesian phylogenetic reconstruction using three mitochondrial gene fragments (12S , 16S and COI) to solve several taxonomic uncertainties among species of cross‐backed grasshoppers (genus Dociostaurus F ieber) from the western M editerranean. P hylogenetic analyses demonstrate the polyphyletic character of subgenera Dociostaurus , Kazakia B ey‐B ienko and Stauronotulus T arbinsky and, thus, the necessity of revising the currently accepted taxonomic subgenera within the genus Dociostaurus . We propose the split of closely related taxa with allopatric distributions such as D. (S.) kraussi and D. (S.) crassiusculus , considering the later a distinct species limited to the I berian P eninsula and excluding the name crassiusculus from other forms of D. (S.) kraussi from E ast E urope and A sia. Estimates of divergence times indicate that diversification of Dociostaurus probably happened during the M iocene–P liocene (3–7 Ma), and the split of the studied pairs of sister taxa took place during the middle and late P leistocene (1–2 Ma). This study highlights the need for more molecular studies on the genus and their different species for a better understanding of their evolution, genetic variation and population dynamics in order to prioritize strategies for their adequate conservation and management.     

New species and biostratigraphic significance of middle Palaeozoic brachiopod genus Linguopugnoides Havliček

Linguopugnoides Havli?ek 1960, originally based on 3 Early Devonian (Lochkovian‐Pragian) species from the Czech Republic, is now construed as embracing at least 25 species distributed over all northern hemisphere continents as well as Australia, and extending from late Llandovery to early Emsian. Species‐diversity peaked during the Lochkovian when the genus was widespread, with 15 species having been reported. Four species are described as new. L. praecarens sp.nov. from the latest Silurian (Pridoli) of the Turkestan Range (South Tien Shan) is considered to be ancestral to the numerous, mainly late Lochkovian‐Pragian forms that appeared during the great diversification of the genus during the early Devonian. Three new species are described from the Lochkovian of the Zeravshan Range, Uzbekistan: L. rectiplicatus, L. diversiplicatus as well as the previously figured but not described L. kimi Rzhonsnitskaya. Species of Linguopugnoides have biostratigraphic and biogeographic significance and have obvious value for inter‐regional correlation.     

Towards a molecular phylogeny of the fungus gnat genus Boletina (Diptera: Mycetophilidae)

Martinsson, S., Kjærandsen, J. & Sundberg, P. (2011). Towards a molecular phylogeny of the fungus gnat genus Boletina (Diptera: Mycetophilidae). —Zoologica Scripta, 40, 272–281. Boletina is a species rich genus of fungus gnats (Diptera: Mycetophilidae) with a mainly Holarctic distribution. The systematics within the genus has gained little attention and this is a first attempt to shed some light over the systematics of Boletina and to test the segregation of the genera Saigusaia and Aglaomyia from Boletina. The nuclear marker 28S and mitochondrial 16S, COI and CytB were amplified and sequenced for 23 taxa that were analysed separately and together with a broad sample of outgroup taxa obtained from GenBank, where also 18S sequences were added. Phylogenies were estimated using maximum likelihood, Bayesian inference and parsimony. We strengthen the hypothesized sister‐group relationship between Docosia and Boletina, but the genus Boletina as currently delimited appears to be paraphyletic and nested in a clade together with Aglaomyia, Coelosia and Gnoriste. The genus Saigusaia, on the other hand, seems to be well separated from Boletina. The Boletina erythropyga species group is consistently found as a distinct basal clade within Boletina s.l. The results obtained are otherwise ambiguous both for the taxa in focus and in some analyses globally with a statistically supported total breakdown of the traditional higher classification into tribes, subfamilies and even families. Interestingly, this breakdown almost disappeared when additional 18S sequences were added.     

Exploring the Afromontane centre of endemism: Kniphofia Moench (Asphodelaceae) as a floristic indicator

Aim The genus Kniphofia contains 71 species with an African–Malagasy distribution, including one species from Yemen. The genus has a general Afromontane distribution. Here we explore whether Kniphofia is a floristic indicator of the Afromontane centre of endemism and diversity. The South Africa Centre of diversity and endemism was explored in greater detail to understand biogeographical patterns. Location Africa, Afromontane Region, southern Africa, Madagascar and Yemen. Methods Diversity and endemism for the genus were examined at the continental scale using a chorological approach. Biogeographical patterns and endemism in the South Africa Centre were examined in greater detail using chorology, phenetics, parsimony analysis of endemicity (PAE) and mapping of range‐restricted taxa. Results Six centres of diversity were recovered, five of which are also centres of endemism. Eight subcentres of diversity are proposed, of which only two are considered subcentres of endemism. The South Africa Centre is the most species‐rich region and the largest centre of endemism for Kniphofia. The phenetic analysis of the South Africa Centre at the full degree square scale recovered three biogeographical areas that correspond with the subcentres obtained from the chorological analysis. The PAE (at the full degree square scale) and the mapping of range‐restricted taxa recovered two and six areas of endemism (AOEs), respectively. These latter two approaches produced results of limited value, possibly as a result of inadequate collecting of Kniphofia species. Only two AOEs were identified by PAE and these are embedded within two of the six AOEs recovered by the mapping of range‐restricted taxa. All the above AOEs are within the three subcentres found by chorological and phenetic analysis (at the full degree square scale) for the South Africa Centre. Main conclusions The centres for Kniphofia broadly correspond to the Afromontane regional mountain systems, but with some notable differences. We regard Kniphofia as a floristic indicator of the Afromontane Region sensu lato. In southern Africa, the phenetic approach at the full‐degree scale retrieved areas that correlate well with those obtained by the chorological approach.     

A new lectin from tulip (Tulipa) bulbs

A lectin was isolated from tulip (Tulipa) bulbs by affinity chromatography on fetuin-agarose and partially characterized. The tulip lectin is a tetrameric protein composed of four identical subunits of M_r 28 000, which are not held together by disulphide bonds. It is not glycosylated and has an amino-acid composition typified by a high content of asparagine-aspartic acid, leucine, glycine and serine. Tulip lectin agglutinates human red blood cells, but has a much higher specific activity with rabbit erythrocytes. In hapten-inhibition assays with the latter type of red blood cell the lectin exhibits a complex specificity, whereas its agglutination with human erythrocytes is readily inhibited by N-acetylgalactosamine, lactose, fucose and galactose.Abbreviations DEAE diethylaminoethyl - PBS phosphate-buffered saline - TL Tulipa lectin - M_r relative molecular mass - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Towards understanding the dynamics of hybridization and apomixis in the evolution of the genus Boechera (Brassicaceae)

We have reviewed past and recent research on the radiation of the North American genus Boechera (Brassicaceae), and discuss future prospects for our understanding of the evolutionary processes and patterns of differentiation in this highly polymorphic genus. Boechera comprises about 80 biennial to perennial species of exclusively North American and Greenlandic distribution. Hybridization and apomixis, in conjunction with polyploidy, were found to play a major role in the origin and maintenance of intraspecific and interspecific polymorphism. However, taxonomic classification within the genus has mainly been based on morphological characters, and diagnostic features discriminating between taxa are often limited. Molecular markers are promising tools for: the assessment of infrageneric phylogen‐etic relationships, the isolation of basal sexual taxa, the detection of hybridization and estimation of its frequency among lineages in time and space, the identification of centres of genetic and species diversity, and the comparison of various reproductive modes. We present and discuss the major achievements obtained since the introduction of molecular methods to Boechera research with respect to these aims, and point out possible shortcomings of specific marker systems. The distribution and transmission of apomixis, polyphyletic origin of genotypes and cytotypes, variation in reproductive system, and the lack of consensus between current taxonomic concepts and evolutionary evidence are the major topics discussed here in the context of hybridization.     

Survival in northern microrefugia in an endemic Carpathian gammarid (Crustacea: Amphipoda)

Gammarus leopoliensis (Crustacea: Amphipoda) is considered a north‐eastern Carpathian endemic species and therefore can be regarded as an appropriate model for testing the hypothesis of Quaternary glacial survival in northern microrefugia. However, 250 km south, the south‐western Carpathians harbour populations that resemble phenotypically both G. leopoliensis and Gammarus kischineffensis, a similar species distributed east of the Carpathians. We used maximum‐likelihood and Bayesian methods to evaluate the phylogenetic relationships of these three taxa based on mitochondrial and nuclear markers, and quantitatively compared diversity patterns, phylogeography and divergence times among north‐eastern and south‐western Carpathian taxa. Results indicate that G. leopoliensis and the south‐western populations form together a strongly supported group (G. leopoliensis s.l.) which, along with G. kischineffensis, belongs to the Gammarus balcanicus clade. This group contains 12 lineages mainly of Pliocene age. G. leopoliensis consists of two widely distributed and recently expanded allopatric sister lineages that diverged from the southern ones ca. 4 Ma, indicating long‐term survival in northern microrefugia. The southern lineages are micro‐endemic and display a scattered distribution, suggesting a more ancient, relict pattern. We conclude that the contrasting diversity patterns between the disjunct distributional areas of G. leopoliensis s.l. reflect differential survival of lineages across the latitudinal gradient, offering a promising system for comparing the evolutionary ecology of lineages persisting in latitudinally disconnected microrefugia. These results fill an important gap in the knowledge of European gammarid biogeography and reveal that all Carpathian Gammarus taxa are ancient and diverse species complexes.     

On the occurrence of tuliposides in the liliiflorae

Approximately 200 samples of liliiflorous plants were investigated for the presence of tuliposides. Appreciable amounts of tuliposide A were detected in all species of Erythronium, Tulipa, Gagea, Bomarea and Alstroemeria. Large amounts of tuliposide B seem to be restricted to Erythronium and Tulipa. The occurrence of identical post-inhibitins in Tulipa and allied taxa and in Alstroemeria and allied taxa is interpreted as indicating a close relationship between Lilioideae and Alstroemeriaceae. At the same time the allergenic potentialities of all taxa of Alstroemeria are stressed.     

Heterochromatin and chromosome variation in cultivated species ofTulipa subg.Eriostemones (Liliaceae)

The chromosomes of several cultivatedTulipa species belonging to the subg.Eriostemones were examined using conventional staining and C-banding techniques. Most of the species have lightly banded chromosomes with heterochromatin content varying from nil to about 15%. The banding patterns of several taxa are described and discussed in regard to species relationships.