Does Type of Substratum Affects Chironomid Larvae Assemblage Composition? A Study in a River Catchment in Northern Patagonia,Argentina

Chironomid larvae assemblages were investigated at seven sampling stations in relation to stream habitat type in the Challhuaco-Ñireco river system located in the Nahuel Huapi National Park, in the North-Western part of Rio Negro Province, Argentina. A total of 2229 individuals were sampled and 43 Chironomidae taxa were recorded with Orthocladiinae (29) being the best represented subfamily, followed by Diamesinae (5), Tanypodinae (3), Podonominae (3) and Chironominae (3). Stictocladius spF, Cricotopus (Paratrichocladius) sp6, Cricotopus sp2, Cricotopus sp3 and Parapsectrocladius sp2 were the most abundant taxa. Sites near the source of the river system were dominated by Podonomus sp., Limnophyes sp., Parapsectrocladius sp. and Stictocladius spF, whereas sites close to the river mouth were dominated by Diamesinae sp5 and Cricotopus sp. Rank/abundance plots show that all the analysed sites displayed dominance of some species. Stictocladius spF, Cricotopus sp2, Cricotopus sp3, Cricotopus (Paratrichocladius) sp6, Parapsectrocladius sp. and Limnophyes sp. resulted as dominant species, or being part of a group of dominant species, at least in one sample. Eleven taxa were associated with habitats related with riffle areas and stable substrates with filamentous algae. Species–environment relationships were examined using ordination analysis. Elevation was the most significant environmental variable that explained 22% of the total variability of the chironomid assemblages, showing stronger relationships among sites within an altitudinal gradient than among habitat type. Abundance of chironomids increased from headwaters to the outflow in Nahuel Huapi Lake responding to an altitude gradient as well as some environmental factors such as coarse matter and nutrient concentrations.     

Navigating the mtDNA road map out of the morphological maze: interpreting morphological variation in the diverse Monomorium rothsteini (Forel) complex (Hymenoptera: Formicidae)

Monomorium is a large and diverse ant genus with speciose radiations in both the Afrotropical and Australian regions. According to the most recent taxonomic revision, many Australian species are characterised by very broad distributions and variable morphology, which suggests that some species may be unrecognised species complexes. With a continent‐wide distribution and diverse yet overlapping morphology, M. rothsteini (Forel) is representative of the greater challenge that exists in Australian Monomorium systematics. Here we investigate species boundaries in M. rothsteini using a molecular phylogenetic framework to interpret the complex overlap of nine morphological characters (with 31 states) and examine biogeographic relationships among the lineages. Bayesian inference resolved 38 mtDNA lineages that were morphologically separable, at least from their sister lineage. Although the morphological characters were intermixed across the phylogeny, instances of inseparable morphology among sister clades was rare. Seventeen lineages exhibited complete morphological overlap with one or more other lineages and could not be separated by Principal Component Analysis based on 12 morphometric variables. Two‐thirds of all lineages occurred sympatrically with one or more both genetically and morphologically divergent lineages. The two nuclear markers EF1αF2 and wingless were used to generate haplotype networks which were characterised by a star‐like pattern indicative of a rapid and recent radiation. Several haplotypes for both nuclear gene regions were shared among individuals occurring in separate mtDNA clades which we were also unable to distinguish morphologically or that were occurring in sympatry, indicating possible introgression in both the mtDNA and nuclear genomes. Clear biogeographic affinities among samples within a lineage were detected but there was no overall pattern in the biogeographic relationships among the lineages. We conclude that M. rothsteini is a large species complex that has undergone a complex evolutionary history following aridification of the Australian continent, and discuss the implications of this conclusion for the systematics of Australian Monomorium more generally.     

Biotic and abiotic drivers of evolution in some Australian thornbills (Passeriformes: Acanthiza) in allopatry,sympatry, and parapatry including a case of character displacement

Disentangling historical, ecological, and abiotic drivers of diversity among closely related species can benefit from morphological diversity being placed in a phylogenetic context. It can also be aided when the species are variously in allopatry, parapatry, and sympatry. We studied a clade of Australian thornbills (Passeriformes: Acanthizidae: Acanthiza) comprising the Brown Thornbill (A. pusilla), Inland Thornbill (A. apicalis), Mountain Thornbill (A. katherina), and Tasmanian Thornbill (A. ewingii) whose distributions and ecology facilitate this approach. We first clarified phylogenetic relationships among them and then detected a low level of gene flow in parapatry between a non-sister pair (Brown, Inland). Further work could partition relative roles of past and current hybridization. We identify likely cases of ecologically driven divergent selection and one of convergent evolution. Divergent selection was likely key to divergence of Inland Thornbills from the Brown–Mountain sister pair. In contrast, convergence in plumage between the non-sister Brown and Inland Thornbills has been driven by their mesic forest habitats on opposite sides of the Australian continent. Finally, morphological distinctiveness of Tasmanian populations of Brown Thornbills could reflect character displacement in sympatry with the ecologically similar Tasmanian Thornbills. Collectively, the combined morphological, genetic, and ecological evidence points to diverse evolutionary processes operating across this closely related group of birds.     

Developments in the taxonomy of silica‐scaled chrysophytes – from morphological and ultrastructural to molecular approaches

Taxonomy in silica‐scaled chrysophytes has gone through three morphological phases. From primary studies of the cell morphology in the 18th century, the focus was in the 20th century replaced by studies of the silica structures of the cell envelope. Now, in the latest decades the importance of DNA sequencing has been recognized, not only to support the taxonomic framework but also to obtain new understanding of taxonomic relations among particular taxa. In the first part of this review, we provide a historical overview of the developments in the taxonomy of scale‐bearing chrysophytes. In the second part, we present a phylogenetic reconstruction of chrysophyte algae, updated by newly obtained SSU rDNA and rbcL sequences of several isolated Synura, Mallomonas and Chrysosphaerella species. We detected significant incongruence between the phylogenies obtained from the different datasets, with the SSU rDNA phylogram being the most congruent with the morphological data. Significant saturation of the first rbcL codon position could indicate the presence of positive selection in the rbcL dataset. Within the Synurales, the relationships revealed by the phylogenetic analyses highlight the artificial infragenetic classification of Mallomonas and Synura, and the occurrence of cryptic diversity within a number of traditionally defined species. Finally, three new combinations are proposed based on the phylogenetic analyses: Tessellaria lapponica, Synura asmundiae and S. bjoerkii.     

DNA evidence for morphological and cryptic Cenozoic speciations in the Anaspididae, ‘living fossils’ from the Triassic

The speciation history of Anaspides tasmaniae (Crustacea: Malacostraca) and its close relatives (family Anaspididae) was studied by phylogenetic and molecular clock analyses of mitochondrial DNA sequences. The phylogenetic analyses revealed that the Anaspides morphotype conceals at least three cryptic species belonging to different parts of its range. The occurrence of multiple cryptic phylogenetic species within one morphological type shows that substantial genetic evolution has occurred independently of morphological evolution. Molecular clock dating of the speciation events that generated both the cryptic and the morphological species of Anaspididae indicated continuous speciation within this group since the Palaeocene ~55 million years ago. This relatively constant rate of recent morphological and cryptic speciation within the Anaspididae suggests that the speciation rate in this group does not correlate with its low extinction rate or morphological conservatism.     

Island life – classification,speciation and cryptic species of Pycnandra (Sapotaceae) in New Caledonia

Pycnandra (Sapotaceae), the largest endemic genus in New Caledonia, comprises 66 species classified in six subgenera. We tested phylogenetic relationships and a proposed infrageneric classification by sampling 60 species for sequences of nuclear ribosomal DNA (ETS, ITS, RPB2) and plastid DNA (trnH–psbA) and nine morphological characters. Data were analysed with Bayesian inference, parsimony jackknifing and lineage through time. We recovered a phylogenetic tree supporting the recognition of six proposed subgenera (Achradotypus, Leptostylis, Pycnandra, Sebertia, Trouettia and Wagapensia). Because a subgeneric classification is used, the nomenclature will be stable when the members are transferred to Pycnandra. Morphological traits were optimized in the BEAST analysis, adding evidence to earlier work that morphology has limited value for successfully diagnosing groups in Sapotaceae. We confirm a previously suspected case of cryptic species that exhibit the same morphological features and require the same abiotic conditions, but are distantly related in the phylogenetic tree. We detected two possible new cases of cryptic sibling species that might warrant recognition. A slowdown in speciation rate in several genera has been suggested as evidence that New Caledonia was once submerged after rifting from Australia. Plotting lineages through time reveals two important intervals at 7.5–8.6 Ma and present to 1.5 Ma, when net molecular diversification within the genus was zero. This indicates that the genus presently has reached a dynamic equilibrium, providing additional evidence that New Caledonia is an old Darwinian island, being submerged during the Eocene and colonized after re‐emergence c. 37 Ma. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 57–77.     

Unmasking cryptic species: morphometric and phylogenetic analyses of the Ibero‐North African Linaria incarnata complex

Linaria incarnata has been treated as a widely distributed Ibero‐North African species in the latest taxonomic reviews. Morphological and phylogenetic disparity between populations of this species has been previously reported. Here we present new morphological and phylogenetic evidence for the disintegration of L. incarnata into three distinct species: L. incarnata from the western Iberian Peninsula; L . mamorensis sp. nov. from north‐western Morocco; and L. onubensis from south‐western Spain. The relatively poor morphological differentiation between these taxa (which can be regarded as cryptic species) and their distinct phylogenetic positions indicate that characters of the L. incarnata morphotype have been acquired multiple times in the evolution of Linaria section Versicolores. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 00, 000–000.     

Prevalence,geographic distribution and phylogenetic relationships among cryptic species of Plasmopara viticola in grape‐producing regions of Georgia and Florida,USA

Previous phylogenetic studies of the grape downy mildew pathogen, Plasmopara viticola, revealed five cryptic species in eastern North America that differed in their host range and geographic distribution. Preliminary comparative studies also documented differences in temperature responses during infection between certain cryptic species, indicating the biological relevance of knowing which cryptic species of the pathogen are present in a given region. However, limited information is available regarding the presence, prevalence and dynamics of cryptic species of P. viticola in the southeastern United States. Here, 301 P. viticola isolates obtained from cultivated grape species in five distinct grape‐growing regions of Georgia and Florida were subjected to cleaved amplified polymorphic sequence analysis and multilocus sequencing (internal transcribed spacer region of the rDNA, actin and β‐tubulin) to identify cryptic species and infer phylogenetic relationships. Three cryptic species, P. viticola clade aestivalis (Pva), clade vinifera (Pvv) and clade vulpina (Pvu), were identified in Georgia, whereas two, Pva and Pvv, were found in Florida; all three cryptic species are reported here for the first time in Georgia, whereas Pva is reported for the first time in Florida. Pva was the most prevalent cryptic species (72.1% of isolates) and was distributed widely from the North Georgia Mountains to Mid‐Florida, whereas Pvv (27.2%) and Pvu (0.7%) were found only in the Coastal Plain region of the two states. Interestingly, Pvu was obtained from French American hybrid Blanc du Bois and could be subcultured on Vitis vinifera Chardonnay, suggesting a broader host range than only the wild species Vitis vulpina reported previously.     

The evolution of climate tolerance in conifer‐feeding aphids in relation to their host's climatic niche

Climate adaptation has major consequences in the evolution and ecology of all living organisms. Though phytophagous insects are an important component of Earth's biodiversity, there are few studies investigating the evolution of their climatic preferences. This lack of research is probably because their evolutionary ecology is thought to be primarily driven by their interactions with their host plants. Here, we use a robust phylogenetic framework and species‐level distribution data for the conifer‐feeding aphid genus Cinara to investigate the role of climatic adaptation in the diversity and distribution patterns of these host‐specialized insects. Insect climate niches were reconstructed at a macroevolutionary scale, highlighting that climate niche tolerance is evolutionarily labile, with closely related species exhibiting strong climatic disparities. This result may suggest repeated climate niche differentiation during the evolutionary diversification of Cinara. Alternatively, it may merely reflect the use of host plants that occur in disparate climatic zones, and thus, in reality the aphid species' fundamental climate niches may actually be similar but broad. Comparisons of the aphids' current climate niches with those of their hosts show that most Cinara species occupy the full range of the climatic tolerance exhibited by their set of host plants, corroborating the hypothesis that the observed disparity in Cinara species' climate niches can simply mirror that of their hosts. However, 29% of the studied species only occupy a subset of their hosts' climatic zone, suggesting that some aphid species do indeed have their own climatic limitations. Our results suggest that in host‐specialized phytophagous insects, host associations cannot always adequately describe insect niches and abiotic factors must be taken into account.     

Species delimitation methods reveal cryptic diversity in the Hypnea cornuta complex (Cystocloniaceae,Rhodophyta)

The simple and convergent morphologies of many red algae make these species difficult to identify using traditional morphological characters. Many cryptic species have been described in recent years based on molecular datasets, and this has led to the application of an integrative taxonomy approach in species delimitation. Here, we performed several species delimitation methods (mBGD, ABGD, SPN, PTP, GMYCs and GMYCm) based on two different loci (COI-5P and rbcL) in species of the Hypnea cornuta complex. These methods were combined with morphological and phylogenetic data, extensive sampling, analysis of topotype material, and historically relevant herbarium samples. Our findings demonstrate that the groups morphologically assigned to H. cornuta and H. stellulifera consist of five different cryptic species. H. cornuta is a polyphyletic taxon composed of three well-separated lineages, thus requiring sequencing of type or topotype specimens to determine which one is Hypnea cornuta sensu stricto. We have revealed that the distribution of H. stellulifera is limited to Asia, while the Brazilian specimens initially assigned to this species were clarified as a new endemic species: Hypnea cryptica sp. nov. Our results indicated that only an integrative approach combining several lines of evidence, including morphology, nomenclature history, molecular data, biogeography and ecology can correctly solve the taxonomic status of widely distributed cryptic species.     

The role of phylogeny and ecology in shaping morphology in 21 genera and 127 species of Australo‐Papuan myobatrachid frogs

Body shape is predicted to differ among species for functional reasons and in relation to environmental niche and phylogenetic history. We quantified morphological differences in shape and size among 98.5% of the 129 species and all 21 genera of the Australo‐Papuan endemic myobatrachid frogs to test the hypothesis that habitat type predicts body shape in this radiation. We tested this hypothesis in a phylogenetic context at two taxonomic levels: across the entire radiation and within the four largest genera. Thirty‐four external measurements were taken on 623 museum specimens representing 127 species. Data for seven key environmental variables relevant to anurans were assembled for all Australian‐distributed species based on species' distributions and 131,306 locality records. The Australo‐Papuan myobatrachid radiation showed high diversity in adult body size, ranging from minute (15 mm snout–vent length) to very large species (92 mm), and shape, particularly sin relative limb length. Five main morphological and environmental summary variables displayed strong phylogenetic signal. There was no clear relationship between body size and environmental niche, and this result persisted following phylogenetic correction. For most species, there was a better match between environment/habitat and body shape, but this relationship did not persist following phylogenetic correction. At a broad level, species fell into three broad groups based on environmental niche and body shape: 1) species in wet habitats with relatively long limbs, 2) species in arid environments with relatively short limbs (many of which are forward or backward burrowers) and 3) habitat generalist species with a conservative body shape. However, these patterns were not repeated within the four largest genera ? Crinia, Limnodynastes, Pseudophryne and Uperoleia. Each of these genera displayed a highly conservative anuran body shape, yet individual species were distributed across the full spectrum of Australian environments. Our results suggest that phylogenetic legacy is important in the evolution of body size and shape in Australian anurans, but also that the conservative body plan of many frogs works well in a wide variety of habitats.     

Molecular phylogeny of hipposiderid bats (Chiroptera: Hipposideridae) from Solomon Islands and Cape York Peninsula,Australia

Throughout the Old World Tropics, examples of cryptic diversity have been demonstrated in leaf‐nosed bats from the Family Hipposideridae. In addition to cryptic diversity, the widespread distributions and graded morphologies of some species render the taxonomy of this family poorly resolved. We used sequences of the mitochondrial gene ND2 and the nuclear gene RAG1 to reconstruct molecular phylogenetic relationships for hipposiderid species. Specimens from Solomon Island and Cape York Peninsula, Australia, were sequenced and combined with published sequences from Southeast Asia. Our results suggest the presence of undescribed diversity in northern Australia. Sequences of ND2 and RAG1 for samples of Cape York Hipposideros ater were distinct from those of Southeast Asia, with corrected sequence divergence estimates for the mitochondrial gene exceeding 23%. In comparison, there was no cryptic diversity detected in Solomon Island specimens when compared to conspecifics from Southeast Asia. On the contrary, three species members of the diadema morphological group from the region exhibit variability in the mitochondrial gene ND2, far below that of other species included in the analyses.     

Cryptic diversity in Brevipalpus mites (Tenuipalpidae)

Defining the taxonomic identity of organisms is a prerequisite for their study, and in the case of economically important species, misidentification may lead to the application of inappropriate prevention and control strategies. Flat mites of the Brevipalpus genus include several crop pests and the systematics of these mites represents a challenge for acarologists. Many of the most economically important Brevipalpus species have repeatedly been inaccurately identified. Such problematic classification has been attributed to the likely occurrence of cryptic species in the genus. In this study, we used an integrative approach that combined molecular analyses, including sequence‐based species delimitation, with detailed morphological identification using traits that have recently showed to be taxonomically informative. Sequences of mitochondrial cytochrome c oxidase subunit I (COI) were obtained from individuals collected from host plants belonging to 14 genera and 13 families across 29 locations in the Americas (Brazil, Chile, USA). The phylogenetic analyses included previously published Brevipalpus sequences from GenBank, and the final data set was classified into 44 haplotypes. Six putative species were recognised by COI‐based species delimitation analysis, and morphological evidence supported each of these species. The integrative approach revealed the occurrence of cryptic species in the Brevipalpus genus and contributed to the clarification of previously noted incongruences. The results presented here allow for the evaluation of taxonomic characteristics in a phylogenetic context and indicate new characters for the differentiation of Brevipalpus species. In addition, Brevipalpus incognitus n. sp. Ferragut & Navia, a cryptic species detected in this study, is described based on morphological and molecular traits. Implications of the advances in Brevipalpus systematics presented herein with respect to pest management are briefly discussed.     

Evolution of the Mitochondrial Cytochrome Oxidase II Gene in Collembola

The sequence of the mitochondrial COII gene has been widely used to estimate phylogenetic relationships at different taxomonic levels across insects. We investigated the molecular evolution of the COII gene and its usefulness for reconstructing phylogenetic relationships within and among four collembolan families. The collembolan COII gene showed the lowest A + T content of all insects so far examined, confirming that the well-known A + T bias in insect mitochondrial genes tends to increase from the basal to apical orders. Fifty-seven percent of all nucleotide positions were variable and most of the third codon positions appeared free to vary. Values of genetic distance between congeneric species and between families were remarkably high; in some cases the latter were higher than divergence values between other orders of insects. The remarkably high divergence levels observed here provide evidence that collembolan taxa are quite old; divergence levels among collembolan families equaled or exceeded divergences among pterygote insect orders. Once the saturated third-codon positions (which violated stationarity of base frequencies) were removed, the COII sequences contained phylogenetic information, but the extent of that information was overestimated by parsimony methods relative to likelihood methods. In the phylogenetic analysis, consistent statistical support was obtained for the monophyly of all four genera examined, but relationships among genera/families were not well supported. Within the genus Orchesella, relationships were well resolved and agreed with allozyme data. Within the genus Isotomurus, although three pairs of populations were consistently identified, these appeared to have arisen in a burst of evolution from an earlier ancestor. Isotomurus italicus always appeared as basal and I. palustris appeared to harbor a cryptic species, corroborating allozyme data. Received: 12 January 1996 / Accepted: 10 August 1996     

Morphological and molecular description of new species of squat lobster (Crustacea: Decapoda: Galatheidae) from the Solomon and Fiji Islands (South‐West Pacific)

The family Galatheidae is among the most diverse families of anomuran decapod crustaceans, and the South‐West Pacific is a biodiversity hot spot for these squat lobsters. Attempts to clarify the taxonomic and evolutionary relationships of the Galatheidae on the basis of morphological and molecular data have revealed the existence of several cryptic species, differentiated only by subtle morphological characters. Despite these efforts, however, relationships among genera are poorly understood, and the family is in need of a detailed systematic review. In this study, we assess material collected in different surveys conducted in the Solomon Islands, as well as comparative material from the Fiji Islands, by examining both the morphology of the specimens and two mitochondrial markers (cytochrome oxidase subunit I, COI, and 16S rRNA). These two sources of data revealed the existence of eight new species of squat lobster, four of which were ascribed to the genus Munida, two to the genus Paramunida, one to the genus Plesionida, and the last species was ascribed to the genus Agononida. These eight species are described along with phylogenetic relationships at the genus level. Our findings support the taxonomic status of the new species, yet the phylogenetic relationships are not yet fully resolved. Further molecular analysis of a larger data set of species, and more conserved genes, will help clarify the systematics of this group. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156 , 465–493.     

DNA barcoding of genus Toxoptera Koch (Hemiptera: Aphididae): Identification and molecular phylogeny inferred from mitochondrial COI sequences

Abstract Identification of aphid species is always difficult due to the shortage of easily distinguishable morphological characters. Aphid genus Toxoptera consists of species with similar morphology and similar to Aphis in most morphological characters except the stridulatory apparatus. DNA barcodes with 1 145 bp sequences of partial mitochondrial cytochrome‐coxidase I (COI) genes were used for accurate identification of Toxoptera. Results indicated mean intraspecific sequence divergences were 1.33%, whereas mean interspecific divergences were greater at 8.29% (0.13% and 7.79% if T. aurantii 3 and T. aurantii 4 are cryptic species). Sixteen samples were distinguished to four species correctly by COI barcodes, which implied that DNA barcoding was successful in discrimination of aphid species with similar morphology. Phylogenetic relationships among species of this genus were tested based on this portion of COI sequences. Four species of Toxoptera assembled a clade with low support in maximum‐parsimony (MP) analysis, maximum‐likelihood (ML) analysis and Bayesian phylogenetic trees, the genus Toxoptera was not monophyletic, and there were two sister groups, such as T. citricidus and T. victoriae, and two clades of T. aurantii which probably presented cryptic species in the genus.     

Astragalus raphaelis (Fabaceae), a critical species from Sicily and taxonomic remarks on A. sect. Sesamei

The taxonomic position of Astragalus raphaelis, a rare and misappreciated species endemic to Sicily, has been examined. In addition, its geographical distribution, ecology and conservation status, diagnostic morphological characters, pod and seed micro‐morphology and relationship with A. stella are provided. Phenetic and phylogenetic trees (based on morphological data) for A. sect Sesamei shows that A. raphaelis is a taxonomically quite isolated species, with some relationships to A. sesamoides.     

Triton's trident: cryptic Neogene divergences in a marine clam (Lasaea australis) correspond to Australia's three temperate biogeographic provinces

The southern coast of Australia is composed of three distinct biogeographic provinces distinguished primarily by intertidal community composition. Several ecological mechanisms have been proposed to explain their formation and persistence, but no consensus has been reached. The marine clam Lasaea australis is arguably the most common bivalve on southern Australian rocky shores and occurs in all three provinces. Here, we tested if this species exhibits cryptic genetic structuring corresponding to the provinces and if so, what mechanisms potentially drove its divergence. Variation in two mitochondrial genes (16S and COIII) and one nuclear gene (ITS2) was assayed to test for genetic structuring and to reconstruct the clam's phylogenetic history. Our results showed that L. australis is comprised of three cryptic mitochondrial clades, each corresponding almost perfectly to one of the three biogeographic provinces. Divergence time estimates place their cladogenesis in the Neogene. The trident‐like topology and Neogene time frame of L. australis cladogenesis are incongruent with Quaternary vicariance predictions: a two‐clade topology produced by Pleistocene Bass Strait land bridge formation. We hypothesize that the interaction of the Middle Miocene Climate Transition with the specific geography of the southern coastline of Australia was the primary cladogenic driver in this clam lineage. Additional in‐depth studies of the endemic southern Australian marine biota across all three provinces are needed to establish the generality of this proposed older framework for regional cladogenesis.