Genetic differentiation of disjunct populations of the antsFormica aquilonia andFormica lugubris in Europe

Summary The speciesFormica aquilonia andF. lugubris of the mound-building red wood ants have a disjunct boreoalpine distribution in Europe. The populations ofF. aquilonia in Finland, Switzerland and the British Isles show little genetic differentiation, whereas the populations ofF. lugubris show considerable differentiation. The Central European populations morphologically identified asF. lugubris can be genetically divided into two groups (here called types A and B). Type B is found in the Alps and the Jura mountains, and is genetically inseparable fromF. aquilonia. Type A lives sympatrically with type B in the Jura mountains and is also found in the British Isles. Sympatry of the two types in the Jura shows that these are separate species. It remains open whether type B is morphologically atypicalF. aquilonia or whether it is a separate species, perhaps with a past history of introgression betweenF. aquilonia andF. lugubris. The gene frequencies in the Finnish populations ofF. lugubris differ from those of both types A and B. Genetic differences withinF. lugubris indicate that the populations have evolved separately for a long time. The social structure ofF. lugubris colonies also shows geographic variation. The nests in Finland and the British Isles seem to be mainly monogynous and monodomous, whereas the nests in Central Europe are polygynous and form polydomous colonies.F. aquilonia has polygynous and polydomous colonies in all populations studied.     

Taxonomy and species boundaries in the coral genus Favia Milne Edwards and Haime, 1857 (Cnidaria: Scleractinia) from Thailand revealed by morphological and genetic data

While Faviidae is a widely and uniformly distributed coral family throughout the Indo-Pacific, the extensive phenotypic plasticity of colony surface and corallite features often confounds the use of macromorphological characters in species identification, and contributes to conflict between traditional classification and molecular analyses of the group. Recent advances in morphological and molecular techniques now provide a suite of methods to re-address coral taxonomy in complex groups, such as that represented by the Faviidae. This study combines morphologic measurements including “3D coordinates landmarks” data with phylogenetic assessments of nuclear (ITS) and mitochondrial (COI-trnM) DNA to assess species boundaries in nine species of Faviidae with para-septothecal walls from Thailand. Strong concordance was found between morphological features and a priori groupings based on both morphospecies and genetically defined groups (ITS and COI-trnM). Favia truncatus was the most well-defined species based on morphological analyses, and it was also shown to be monophyletic using phylogenetic analyses. Besides F. truncatus, the only other species that was found to be monophyletic in analyses of both genes was F. cf. helianthoides, but its skeletal morphology overlapped with the F. favus species complex (comprised of F. favus, F. speciosa, F. matthaii and F. rotumana). Although not genetically monophyletic, the F. favus species complex and F. pallida were fairly well delineated morphologically. Morphospecies within the F. favus species complex are therefore possibly a result of genetic drift and/or stable polymorphisms driven by divergent selection. These results represent a first step toward a taxonomic revision of the Indo-Pacific Favia, which will integrate morphological methods with the study of type material, genetic information, reproductive data, and tests of phenotypic plasticity—given that multiple lines of evidence are needed to resolve ambiguous species and assign species names.     

Systematics of Pipizini and taxonomy of European Pipiza Fallén: molecular and morphological evidence (Diptera,Syrphidae)

Vuji?, A., Ståhls, G., A?anski, J., Bartsch, H., Bygebjerg, R. & Stefanovi?, A. (2013). Systematics of Pipizini and taxonomy of European Pipiza Fallén: molecular and morphological evidence (Diptera, Syrphidae). —Zoologica Scripta, 42, 288–305. In the present work the monophyly and molecular phylogenetic relationships of the genera of tribe Pipizini (Syrphidae) were investigated based on mitochondrial cytochrome c oxidase subunit I (COI) and nuclear 28S rDNA sequences, and the relationships among species of genus Pipiza Fallén, 1810 based on mtDNA COI sequences. Molecular phylogenetic analyses of Pipizini supported Pipiza as monophyletic and as sister group to all other Pipizini, and resolved other Pipizini genera as monophyletic lineages except for genus Heringia Rondani, 1856. To recognize the distinctness and maintain the monophyly the genus Heringia was redefined, generic rank was assigned to Neocnemodon Goffe, 1944 stat. n., and the genus Claussenia Vuji? & Ståhls gen. n., type‐species Claussenia hispanica (Strobl, 1909), was described. A revision of the European Pipiza species, including a discussion of taxonomic characters and a morphological redefinition of all included species, is presented. One new species, Pipiza laurusi Vuji? & Ståhls sp. n. was described. The taxa Pipiza carbonaria Meigen, 1822; Pipiza fasciata, Meigen 1822; Pipiza lugubris (Fabricius, 1775), Pipiza noctiluca (Linneaues, 1758), Pipiza notata Meigen, 1822 were redefined. Lectotypes are designated for 17 taxa, and neotypes were designated for seven taxa. Fourteen new synonymies were proposed. Male genitalia were illustrated for all the species, and a key of the 12 European species for males and females was provided. Geometric morphometrics of wing landmarks and extended sampling of mtDNA COI sequences was employed to delimitate taxa of the P. noctiluca and P. lugubris complexes. Despite subtle morphological differences, wing geometric morphometrics variables of wing size and shape showed highly significant differences among species within P. noctiluca and P. lugubris complexes, which were supported by the molecular data.     

RETRACTED ARTICLE: Interspecific competition among ants in the boreal forest: Testing predictions from a linear hierarchical competition model

Competitive interactions often play an important role in local community structure, and particularly so in ant communities. We test predictions derived from a competition model proposed for Scandinavian ant communities by comparing ant nest densities inside and outside fifty randomly-selected territories of competitively dominant wood ants within 30 km of Ume?, northern Sweden. As predicted by the model, competitively-intermediate encounter species, as well as other territorial species, showed complementary occurrences with both studied wood ants, Formica aquilonia Yarrow and F. lugubris Zett.. In contrast, complementary abundances, i.e. lower density of competitively-inferior submissive species in the presence of wood ants, as predicted by the model, was not supported for all submissive species. Of the two studied wood ant species, the nest density of submissive species was negatively correlated only with F. aquilonia. Submissives as a group, as well as Myrmica-species, showed complementary abundances with F. aquilonia, but one Leptothorax and two Serviformica species had higher nest densities in the presence of this competitively-dominant species. We propose that, for Leptothorax, these deviations from the model predictions may be because of limited niche overlap with dominant wood ants, a small worker force and a timid behaviour which does not elicit aggression in wood ants. For the two Serviformica species, we propose a combination of protection against social parasites, inter-specific social control performed by F. aquilonia, and dominance relationships between competitively-inferior submissive species as reasons for the higher nest density inside F. aquilonia territories. Monogyny, and thereby smaller nests, lower worker force, smaller territory (as shown by this study) in F. lugubris, as compared to the polygyny in F. aquilonia, may also help explain the differences in their effects on subordinate species. Our analyses indicate that the linear competition hierarchy model proposed for Scandinavian ants accurately predicts the outcome of interaction between and community composition for dominant territorial and encounter species, but that it needs refinement with respect to the relationship between territorial and submissive species and the resulting community composition. Further studies are needed, especially addressing the complex relationships between these latter groups, and the effects of different competitively-dominant wood ants, to determine the mechanisms determining the outcome of these relationships and to more accurately predict community composition. Received 11 June 2007; revised 11 September 2007; accepted 17 September 2007.     

Occurrence of the marbled rose-chafer (Protaetia lugubris Herbst, Coleoptera, Cetoniidae) in rural avenues in northern poland

The occurrence of Protaetia lugubris, an endangered species developing in hollow trees, was studied in a network of rural avenues in northern Poland. We detected 1002 trees from nine species with hollows suitable for beetle development (25% of all trees inspected). Among them, 74 trees (7.4%) from seven species were occupied by P. lugubris. The distribution of P. lugubris was random with respect to tree species identity. The beetle preferred trees above 200 cm in circumference with a tendency towards higher occupancies of the bigger trees having circumferences above 300 cm. P. lugubris did not show any significant preferences according to hollow entrance area, exposition and road surface type. Our results indicate that P. lugubris is a generalist species colonizing all suitable hollows. Its occurrence indicates suitable conditions for many other species associated with tree cavities and decaying wood.     

Phylogenetic relationships among <Emphasis Type="Italic">Fagopyrum</Emphasis> species based on analysis of the <Emphasis Type="Italic">nad1</Emphasis> Gene b/c intron

For the first time, the b/c intron of the nad1 gene was characterized in 13 species of Fagopyrum and the applicability of the intron was demonstrated for identification of buckwheat species. The length of the b/c intron of the nad1 gene varied from 1217 bp in F. tataricum and F. cymosum to 1239 bp in F. capillatum. Species-specific SNPs and indels were identified. The genus phylogeny that was determined with the molecular approach correlated with phylogenetic data that are based on morphological characteristics. The NJ and MP dendrograms revealed distinct division of the analyzed species into two main groups, cymosum and urophyllum. The species of the urophyllum group were characterized by the presence of several synapomorphic substitutions and indels, including the insertion of 11 nucleotides, which is flanked by direct repeats.     

Numerical taxonomy of some taxa of the genus Fucus in the Iberian Peninsula

A study of morphological variation patterns in three Iberian Fucus species has been carried out. Data on 36 features from 113 specimens have been studied by numerical methods (PCA, discriminant analysis and ANOVA).The PCA ordination displayed a continuous trend of variation in the following Fucus species: F. ceranoides, F. vesiculosus and F. spiralis. However, no obvious morphological discontinuities have been observed. In addition, three varieties of F. spiralis and five of F. vesiculosus have been identified.At the varietal level discriminant functions could be used for identifying 93–100% of the individuals within F. vesiculosus and 72–91% of F. spiralis.The most discriminating diagnostic character appeared to be the midrib size, as detected by ANOVA procedures.Some qualitative characters, such as presence of vesicles, dioecious-monoecious character and receptacle edge shape, frond and receptacle shape, have been also important for discriminating specific and infraspecific taxa.A nomenclatural discussion of varietal taxa is also included.     

Daphnia Galeata × D. Longispina Hybrids in Western Norway

We describe the occurrence of D. galeata×longispina hybrids in two lakes of western Norway. Parental species and interspecific hybrids were characterised by both nuclear and mitochondrial molecular markers. In one of the populations, hybrids were shown to dominate the population over several years. A few individuals in both populations were probably not F1 hybrids, but possibly backcrosses or F2 hybrids. Most (possibly all) F1 hybrids were of D. galeata maternal origin. In addition, interspecific hybrids could be identified based on morphological characters, which were intermediate between the parental species. Interspecific hybridisation between these two species is remarkable, since they are distantly related.     

Unanticipated population structure of European grayling in its northern distribution: implications for conservation prioritization

Background

We compared here the suitability and efficacy of traditional morphological approach and DNA barcoding to distinguish filarioid nematodes species (Nematoda, Spirurida). A reliable and rapid taxonomic identification of these parasites is the basis for a correct diagnosis of important and widespread parasitic diseases. The performance of DNA barcoding with different parameters was compared measuring the strength of correlation between morphological and molecular identification approaches. Molecular distance estimation was performed with two different mitochondrial markers (coxI and 12S rDNA) and different combinations of data handling were compared in order to provide a stronger tool for easy identification of filarioid worms.

Results

DNA barcoding and morphology based identification of filarioid nematodes revealed high coherence. Despite both coxI and 12S rDNA allow to reach high-quality performances, only coxI revealed to be manageable. Both alignment algorithm, gaps treatment, and the criteria used to define the threshold value were found to affect the performance of DNA barcoding with 12S rDNA marker. Using coxI and a defined level of nucleotide divergence to delimit species boundaries, DNA barcoding can also be used to infer potential new species.

Conclusion

An integrated approach allows to reach a higher discrimination power. The results clearly show where DNA-based and morphological identifications are consistent, and where they are not. The coherence between DNA-based and morphological identification for almost all the species examined in our work is very strong. We propose DNA barcoding as a reliable, consistent, and democratic tool for species discrimination in routine identification of parasitic nematodes.     

Morphometric and molecular differentiation of Phlebotomus (Phlebotomus) sandflies

The closely related sandfly species of the subgenus Phlebotomus namely, Phlebotomus papatasi (Scopoli, 1786), Phlebotomus duboscqi Neveu‐Lemair, 1906 and Phlebotomus bergeroti Parrot, 1934 (Diptera: Psychodidae), are major vectors of Leishmania major (Kinetoplastida: Trypanosomatidae), the causative agent of cutaneous leishmaniasis in the Old World. Although allopatric in most of their distribution, the three species exist sympatrically in many places in central and eastern Sudan. Males of the three species can be distinguished using morphological characters; however, females are much harder to identify, thus complicating epidemiological studies. We carried out a morphometric and a molecular study to determine reliable morphological features and develop a polymerase chain reaction (PCR) assay for distinguishing females of these species. Males and females from each species were collected from sites in Sudan, East Africa and from one site in Mali, West Africa. Males were analysed morphologically and 20 characters and 10 character ratios were used in a stepwise discriminant analysis. This led to the identification of four characters with high discriminant loading scores sufficient for accurate male species identification. Male DNA was then used for the development of a PCR‐based species diagnostic based on the second internal transcribed spacer (ITS2) of the ribosomal DNA. A set of four primers was developed to generate fragment sizes that are specific to each species and can reliably identify females as well as hybrid DNA. Both the morphometric and the molecular findings of this study have important applications for studies of the epidemiology of cutaneous leishmaniasis.     

Analysis of reticulate relationships within the Daphnia longispina species complex. Allozyme phenotype and morphology

A two-step method is proposed to get reliable associations between morphology and genotype in clonal assemblages in which more than two predominantly parthenogenetic species are thought to coexist with hybrids. In dataset 1, the genetic relationships among clones of the Daphnia longispina hybrid complex from seven prealpine lakes in southern Germany were studied based on the variation at 21 enzyme loci. The spatial arrangement in the multidimensional scaling plot revealed a reticulate pattern among three presumably parental species, D. cucullata, D. galeata and D. hyalina, and three hybrid groups, D. cucullata/galeata, D. cucullata/hyalina and D. galeata/hyalina. The Got1 locus was believed to be a discriminating factor between species and hybrids (cf. Wolf and Mort, 1986). However, this locus is more variable, and 57% of the clones would have been misidentified using it. Moreover, the morphological variation within genetically defined groups is also higher than previously assumed. In dataset 2, the revision of morphological and genetic markers greatly improved the association between morphology and genotype in newly collected animals. The spatial arrangement of clones in multidimensional scaling plots and morphological asymmetries to parents suggest both, different degrees of introgression and bidirectional hybridization. Most unexpected genotypes were found in the cxh hybrid group, suggesting that F1-hybrids are fertile. The results showed (1) that the clonal diversity was very high (2) that detailed analyses of multiple morphological and allozyme markers are necessary to resolve taxonomic relationships within clonal assemblages consisting of multiple species, hybrids and differently introgressed backcrosses, and (3) that the three original species seem to have sufficient within-species recombination and a low enough rate of backcrossing to allow taxonomic identification. It must remain undecided if the present situation is locally restricted, if it is stable or represents a transient situation which could lead to either a consolidation of the three species by gradual elimination of the hybrids, to a taxonomic breakdown, or to hybrid speciation.     

MOLECULAR PHYLOGENY OF SELECTED SPECIES OF THE ORDER DINOPHYSIALES (DINOPHYCEAE)—TESTING THE HYPOTHESIS OF A DINOPHYSIOID RADIATION1

Almost 80 years ago, a radiation scheme based on structural resemblance was first outlined for the marine order Dinophysiales. This hypothetical radiation illustrated the relationship between the dinophysioid genera and included several independent, extant lineages. Subsequent studies have supplied additional information on morphology and ecology to these evolutionary lineages. We have for the first time combined morphological information with molecular phylogenies to test the dinophysioid radiation hypothesis in a modern context. Nuclear‐encoded LSU rDNA sequences including domains D1‐D6 from 27 species belonging to Dinophysis Ehrenb., Ornithocercus F. Stein, Phalacroma F. Stein, Amphisolenia F. Stein, Citharistes F. Stein, and Histioneis F. Stein were obtained from the Indian Ocean. Previously, LSU rDNA has only been determined from one of these. In Bayesian analyses, Amphisolenia formed a long basal clade to the other dinophysioids. These diverged into two separate lineages, the first comprised species with a classical Phalacroma outline, also including the type species P. porodictyum F. Stein. Thus, we propose to reinstate the genus Phalacroma. The relationship between the genera in the second lineage was not well resolved. However, the molecular phylogeny supported monophyly of Histioneis and Citharistes and showed the genus Dinophysis to be polyphyletic and in need of a taxonomic revision. Species of Ornithocercus grouped with Citharistes, but this relationship remained unresolved. The phylogenetic trees furthermore revealed convergent evolution of several morphological characters in the dinophysioids. According to the molecular data, the dinophysioids appeared to have evolved quite differently from the radiation schemes previously hypothesized. Four dinophysioid species had identical LSU rDNA sequences to other well‐established species.     

New insight into the identification and molecular phylogeny of dagger nematodes of the genus Xiphinema (Nematoda: Longidoridae) with description of two new species

The genus Xiphinema constitutes a large group of about 260 species of plant‐ectoparasitic nematodes. The group is polyphagous and distributed almost worldwide. Some of the species of this genus damage agricultural crops by direct feeding on root cells as well as by transmitting nepoviruses. Species discrimination in Xiphinema is complicated by phenotypic plasticity leading to potential misidentification. We conducted nematode surveys in cultivated and natural environments in Spain from 2009 to 2012, from which we identified 20 populations of Xiphinema species morphologically close to the virus‐vector nematode species Xiphinema diversicaudatum, three apomictic populations tentatively identified as species from the complex Xiphinema aceri‐pyrenaicum group, and one population morphologically different from all others that is characterized by a female tail elongate to conical and absence of uterine differentiation. We developed comparative multivariate analyses for these related species by using morphological and morphometrical features together with molecular data from nuclear ribosomal DNA genes [D2‐D3 expansion segments of large ribosomal subunit 28S, internal transcribed spacer 1 (ITS1), and partial small ribosomal subunit (18S)]. The results of multivariate, molecular, and phylogenetic analysis confirmed the morphological hypotheses and allowed the delimitation and discrimination of two new species in the genus described herein as Xiphinema baetica sp. nov. and Xiphinema turdetanensis sp. nov. , and ten known species: Xiphinema adenohystherum, Xiphinema belmontense, Xiphinema cohni, Xiphinema coxi europaeum, Xiphinema gersoni, Xiphinema hispidum, Xiphinema italiae, Xiphinema lupini, Xiphinema nuragicum, and Xiphinema turcicum. Multivariate analyses based on quantitative and qualitative characters and phylogenetic relationships of Xiphinema spp. based on the three molecular ribosomal markers resulted in a partial consensus of these species grouping as nematode populations were maintained for the majority of morphospecies groups (e.g. morphospecies groups 5 and 6), but not in some others (e.g. position of Xiphinema granatum), demonstrating the usefulness of these analyses for helping in the diagnosis and identification of Xiphinema spp. The clade topology of phylogenetic trees of D2‐D3 and partial 18S regions in this study were congruent in supporting the polyphyletic status of some characters, such as the female tail shape and the degree of development of the genital system in species with both genital branches equally developed. This is the most complete phylogenetic study for Xiphinema non‐americanum‐group species. Agreement between phylogenetic trees and some morphological characters (uterine spines, pseudo‐Z organ, and tail shape) was tested by reconstruction of their histories on rDNA‐based trees using parsimony and Bayesian approaches. Thus, integrative taxonomy, based on the combination of multivariate, molecular analyses with morphology, constitutes a new insight into the identification of Xiphinema species. © 2013 The Linnean Society of London     

Molecular phylogeny of fig wasp pollinators (Agaonidae, Hymenoptera) of Ficus section Galoglychia

The obligate mutualism between fig trees and their fig wasp pollinators, together with the general tendency for each host species to be pollinated by one fig wasp species, led to the hypothesis that these two lineages have cospeciated. The pollinators of African figs of section Galoglychia form a diverse group of genera whose species seem to be less constrained to a specific host than other pollinating fig wasp genera. Various authors have suggested remarkably different phylogenetic relationships between the seven genera associated with section Galoglychia. These uncertainties concerning the classification make it difficult to understand the historical patterns of association between these wasps and their hosts. The phylogenetic tree for the pollinators was reconstructed with 28S, COI and ITS2 DNA sequence data and compared with morphological classification of the hosts. Pollinator genera were monophyletic in all analyses. However, the relative position of some genera remains unresolved. Investigation of host−fig association suggests that there have been frequent host jumps between host subsections. This indicates that cospeciation between fig trees and fig wasps is not as stringent as previously assumed. In addition, pollinators of the genus Alfonsiella associated with three host figs (Ficus craterostoma, F. stuhlmannii and F. petersii) are morphologically very similar in South Africa. We investigated the possibility that these pollinators form a complex of species with host‐based genetic differentiation. Molecular analyses supported the distinction of the pollinator of F. craterostoma as a good species, but the pollinators of F. stuhlmannii and F. petersii clustered within the same clade, suggesting that these two host species share a single pollinator, Alfonsiella binghami. Based on both molecular data and morphological re‐evaluation, a new Alfonsiella species is described, Alfonsiella pipithiensis sp. nov., which is the pollinator of F. craterostoma in southern Africa. A key to both females and males of all described species of Alfonsiella is provided.     

Differentiating three Indian shads by applying shape analysis from digital images

Tenualosa ilisha, T. toli and Hilsa kelee are three Indian shad species of the family Clupeidae. In past, relationships among these shads have been explored in few morphological studies but still remain poorly understood. In this study, 120 specimens of three shad species were collected from the wild and a truss network system of 13 landmarks was interconnected to yield 77 size-adjusted characters. Discriminant function analysis (DFA) produced 13 discriminating variables in delineating the three species. The landmark based morphometric distance between origin of dorsal fin and posterior end of eye was the most important character in discriminating the three species. Three prominent characters extracted from DFA (distance between posterior end of operculum and posterior end of eye, distance between origin of dorsal fin and posterior end of eye and distance between insertion of pelvic fin and end of operculum) were used as traditional morphometric characters and their ratio to standard length were found significant in differentiating the three shad species. The present study strongly supports the efficiency of landmark-based morphological characters generated from digital images in discriminating the species and can be used as a rapid diagnostic tool in recognizing them.