Nuclear rDNA Research Articles

First phenotypic and genotypic description of Fasciola hepatica infecting highland cattle in the state of Mexico, Mexico

Fascioliasis is a plantborne and zoonotic parasitic disease caused by fasciolid liver flukes. Fasciola hepatica is the only fasciolid species described in the Americas. Human fascioliasis endemic areas are mainly located in high altitude areas of the Americas. Given the necessity to characterize F. hepatica populations involved, the phenotypic and genotypic features of fasciolid adults infecting cattle in the highland area of Toluca, State of Mexico, Mexico, were analyzed and compared to fasciolid materials from the Northern Bolivian Altiplano, representing the altiplanic transmission pattern in a hyperendemic scenario. A computer image analysis system (CIAS) was applied on the basis of standardized measurements. The aforementioned F. hepatica highland populations were compared to standard lowland natural populations of European origin (Spain and France) and F. gigantica of African origin (Burkina Faso). Liver-fluke size was studied by principal component analysis (PCA). Two phenotypic patterns could be distinguished in the F. hepatica material analyzed from the Americas: the valley pattern (Toluca, Mexico) and the altiplanic pattern (Northern Altiplano, Bolivia). PCA showed that the Altiplano population presented a large body size range with a pronouncedly lower minimum size. Mahalanobis distances demonstrated that American populations are very close to European populations. Genetic haplotyping was performed using the ribosomal DNA intergenic region, including ITS-1, 5.8S and ITS-2. The intergenic region was 951 bp-long, providing 2 combined haplotypes due to one mutation appearing in the ITS-2 sequence. Molecular results showed that Fh-1A and Fh-2A, the most frequent haplotypes of F. hepatica from southern Europe, are present in Mexican cattle. Nuclear rDNA biomarkers correlated with adult fluke phenotypic characteristics. Results showed that the Mexican population analyzed and European standard populations presented a phenotypic and genotypic homogeneity, suggesting an introduction with livestock transported during the early colonization period. Results are moreover analyzed in terms of altitude and permanent/seasonal transmission characteristics.

Collapsimycopappus: A new leaf pathogen with mycopappus‐like propagules in Mycosphaerellaceae

AbstractA previously undescribed mycopappus‐like fungus associated with large zonate leaf spot disease of Styrax obassia was found in Japan. Single‐propagule isolation of the fungus from necrotic areas of the diseased leaves yielded four pure cultures. Pathogenicity tests of the fungus were performed on potted S. obassia plants. Leaves were wounded with a sterilized needle and inoculated with mycelial agar. Development of necrotic zones was observed within 4 months. Numerous propagules appeared on the necrotic areas, and two strains of the same fungus were isolated again. The morphology of the pathogen was described, and its phylogeny was analysed based on the sequences of nuclear rDNA ITS, LSU regions and the rpb2 genes. Twenty‐four sequences were generated from six strains. Phylogenetic analyses indicated that the pathogen was closely related to the members of Mycosphaerellaceae, but was distinct in having brown propagules with hyphal appendages that collapsed at maturity and divided into part spores. Therefore, we propose Collapsimycopappus as a new genus to accommodate a single species, C. styracis.

Racocetra crispa (Glomeromycotina) delimited by integrative evidence based on morphology, long continuous nuclear rDNA sequencing and phylogeny

Here, we describe a new ornamented arbuscular mycorrhizal (AM) fungus, Racocetra crispa sp. nov. isolated from maize fields from the central region of Minas Gerais State, Brazil. For the first time, a Glomeromycotina species is described using a long continuous nuclear rDNA sequence fragment, which encompasses the nearly complete 18S SSU sequence gene until the 3′ end of the D2 region of the 28S LSU (~ 3100 bp), which allows for comparison with sequences obtained from regions used for fungal metagenomic studies, species description, and AM fungi DNA-barcode. The new species forms dark brown to black spores, approx. 340–510 μm on in diam., on sporogenous cells. The spores have unique “cloud/flower” projections on the spore surface, two walls, and differentiate a multiple-lobed germ shield with up to 8–12 germ tube initiations. The analysis of the intra- and interspecific DNA-barcode sequence variation within the Racocetra showed that the intragenomic polymorphism among the clones of R. crispa (0–2 %) is within the lower range for the genus. The V3–V4 region of the SSU nrDNA has no resolution to discriminate Racocetra at species level, but from this fragment, we found homology between R. crispa and environmental sequences from two metagenomics studies, one carried out in Brazil at the fungus type location and the other in New Zealand. The integration between AM fungal sequences from reference strains and those obtained from environmental sequences in Glomeromycotina is still a problematic issue, mainly due to the reduced number of AM fungal species characterized based on DNA sequences.

Morphological characters and phylogenetic analysis reveal a new species of Phellinus with hooked hymenial setae from Vietnam

Phellinus vietnamensis sp. nov. is described from Vietnam based on morphological characters and molecular data. Morphologically, it is characterized by perennial, pileate basidiomata, a dimitic hyphal system, hooked hymenial setae, and colorless, broadly subglobose to ovoid, thick-walled basidiospores 5.5–6 × 4.8–5.2 μm. Phylogenetically, the status of Phellinus vietnamensis is strongly supported based on sequences of the nuclear internal transcribed spacer (ITS) regions, the translation elongation factor 1-α gene (EF1-α) nuclear large subunit rDNA (nrLSU) and the second largest subunits of RNA polymerase II (RPB2).

Tying up loose threads: revised taxonomy and phylogeny of an avian-dispersed Neotropical rhizomorph-forming fungus

Rhizomorpha corynecarpos Kunze was originally described from wet forests in Suriname. This unusual fungus forms white, sterile rhizomorphs bearing abundant club-shaped branches. Its evolutionary origins are unknown because reproductive structures have never been found. Recent collections and observations of R. corynecarpos were made from Belize, Brazil, Ecuador, Guyana, and Peru. Phylogenetic analyses of three nuclear rDNA regions (internal transcribed spacer, large ribosomal subunit, and small ribosomal subunit) were conducted to resolve the phylogenetic relationship of R. corynecarpos. Results show that this fungus is sister to Brunneocorticium bisporum—a widely distributed, tropical crust fungus. These two taxa along with Neocampanella blastanos form a clade within the primarily mushroom-forming Marasmiaceae. Based on phylogenetic evidence and micromorphological similarities, we propose the new combination, Brunneocorticium corynecarpon, to accommodate this species. Brunneocorticium corynecarpon is a pathogen, infecting the crowns of trees and shrubs in the Neotropics; the long, dangling rhizomorphs with lateral prongs probably colonize neighboring trees. Longer-distance dispersal can be accomplished by birds as it is used as construction material in nests of various avian species.

Molecular phylogeny of the genus Corchorus (Grewioideae, Malvaceae s.l.) based on nuclear rDNA ITS sequences

A molecular phylogenetic analysis of the genus Corchorus (Grewioideae, Malvaceae s.l.) is presented, based on sequences of the nuclear ribosomal DNA internal transcribed spacer (ITS) region for 144 accessions representing 47 species. Several other genera from the subfamily Grewioideae, namely Pseudocorchorus, Triumfetta, Sparrmannia, Entelea, and Grewia, were included as outgroups. The monophyly of the genus was well supported by all phylogenetic analyses (maximum likelihood, Bayesian approaches, and parsimony), and Corchorus was divided into four major clades. The majority of African species formed a statistically highly supported and distinct clade separated from the other pantropically distributed species. Several endemic species from Australia, New Caledonia, and tropical America were nested within this distinct clade, indicating dispersal from Africa to the rest of the pantropics. Based on the taxa included in this study, the two cultivated species (C. olitorius and C. capsularis) shared a common ancestry with wild species of C. africanus, C. brevicornatus, C. pseudocapsularis, C. pseudo-olitorius, C. urticifolius, C. pilosus, C. orinocensis, and C. cunninghamii. Pseudocorchorus, previously considered an accepted genus, was nested within the genus Corchorus and shared a common ancestry especially with C. depressus and C. siliquosus. Based on morphological and anatomical similarity as well as the results of the present molecular findings, inclusion of the six Pseudocorchorus species into Corchorus is proposed, with Pseudocorchorus as a synonym of Corchorus. Of the included outgroup taxa, Triumfetta is the closest sister to Corchorus, while the common ancestor of Corchorus/Pseudocorchorus, Triumfetta, Sparrmannia, and Entelea is Grewia. A further phylogenetic study with more taxa mainly from Australia, together with additional molecular markers and morphological investigation, would help to test the hypothesis on the biogeography and growth form evolution of the genus Corchorus.

Two novel myxosporean parasites in Black Sea fishes: Kudoa niluferi sp. nov. and Kudoa anatolica sp. nov. (Cnidaria: Myxosporea).

Members of the genus Kudoa are typically histozoic and only a few are coelozoic parasites, mainly in marine fishes. In the present study, 2 novel Kudoa species were recovered and described as Kudoa niluferi sp. nov. in the musculature of Neogobius melanostomus and Kudoa anatolica sp. nov. in the musculature, urinary bladder and kidney of Atherina hepsetus collected from the coast of Sinop on the Black Sea. Means ± SD (ranges) of mature spores of K. niluferi sp. nov. were 5.9 ± 0.1 (5.7-6.1) µm in length, 9.2 ± 0.2 (8.8-9.5) µm in width and 7.5 ± 0.3 (7.0-8.1) µm in thickness, while those of K. anatolica sp. nov. were 4.1 ± 0.3 (3.5-4.1) µm in length, 7.1 ± 0.2 (6.7-7.2) µm in width and 5.7 ± 0.2 (5.3-6.0) µm in thickness. In both parasite species, length and width of the 4 polar capsules were not equal and formed 3 distinct size classes, largest (1), intermediate (2) and smallest (1) in size. The prevalence and intensity of infection by K. niluferi sp. nov. were 12.8% and 20-29 parasites (per field-of-view, at 200× magnification), respectively, in the musculature of N. melanostomus. These values for K. anatolica sp. nov. were 32.1% and 10-19 parasites in the musculature as well as 2.9% and 20-29 parasites jointly in the kidney and urinary bladder of A. hepsetus. Phylogenetic analysis based on nuclear small subunit rDNA also suggested K. niluferi and K. anatolica as 2 novel species. These species appeared in the same lineage with K. nova and formed a Black Sea lineage.

Editorial Board

Lophiostoma bipolare was taxonomically revised based on the morphological observations and phylogenetic analyses of molecular data from nuclear rDNA SSU-ITS-LSU, TUB, tef1, and rpb2 genes. Twenty-nine strains were morphologically similar to Lo. bipolare. A total of 174 sequences were generated from the Lo. bipolare complex. Phylogenetic analyses based on TUB sequence revealed 11 distinct species within the Lo. bipolare complex. Morphological features of the ascospores and the anatomical structure of the ascomata from both field collections as well as axenic culture, which have been reported previously as variable features at intraspecific levels, were compared to evaluate the taxonomic reliability of these features. To clarify the generic position of the 11 species, phylogenetic analyses were done on SSU-ITS-LSU-tef1-rpb2 gene sequences. The Lo. bipolare complex shared phylogenetic relationships with Pseudolophiostoma and Vaginatispora, and formed an additional five distinct clades from other members of Lophiostomataceae. According to its phylogenetic position, Lo. bipolare sensu stricto was distantly related to Lophiostoma s. str., and formed an independent clade within Lophiostomataceae. Lophiostoma bipolare s. str. could be distinguished from the other lophiostomataceous genera by the clypeus around the ostiolar neck and by the thin and uniformly thick peridium. A novel genus described as Lentistoma was established to accommodate this species, and the epitypification of Lentistoma bipolare (basionym: Massarina bipolaris) was proposed. Other lineages of the Lo. bipolare complex could not be separated on the basis of the ascospore size and sheath variations, but were distinguished based on ascomatal features, such as the existence of the clypeus, brown hyphae surrounding the peridium, and the contexture of the peridium, which were stable indicators of generic boundaries in Lophiostomataceae. Four additional new genera with five new species were recognised based on these morphological differences: Crassiclypeus (C. aquaticus), Flabellascoma (F. cycadicola and F. minimum), Leptoparies (Lep. palmarum), and Pseudopaucispora (Pseudop. brunneospora). Three new species were added to Pseudolophiostoma (Pseudol. cornisporum, Pseudol. obtusisporum, and Pseudol. tropicum) and two new species were added to Vaginatispora (V. amygdali and V. scabrispora). The re-evaluation of the validity of several previously recognised genera resulted in the introduction of two new genera with new combinations for Lophiostoma pseudoarmatisporum as Parapaucispora pseudoarmatispora and Vaginatispora fuckelii as Neovaginatispora fuckelii.

Assessment of chemical and genetic variability in Tanacetum gracile accessions collected from cold desert of Western Himalaya.

Genetic diversity is essential for survival and adaptation of high altitude plants such as those of Tanacetum genus, which are constantly exposed to environmental stress. We collected flowering shoots of ten accessions of Tanacetum gracile Hook.f. & Thomson (Asteraceae) (Tg 1-Tg 10), from different regions of cold desert of Western Himalaya. Chemical profile of the constituents, as inferred from GC-MS, exhibited considerable variability. Percentage yield of essential oil ranged from 0.2 to 0.75% (dry-weight basis) amongst different accessions. Tg 1 and Tg 6 were found to produce high yields of camphor (46%) and lavandulol (41%), respectively. Alpha-phellendrene, alpha-bisabool, p-cymene and chamazulene were the main oil components in other accessions. Genetic variability among the accessions was studied using RAPD markers as well as by sequencing and analyzing nuclear 18S rDNA, and plastid rbcL and matK loci. The polymorphic information content (PIC) of RAPD markers ranged from 0.18 to 0.5 and the analysis clustered the accessions into two major clades. The present study emphasized the importance of survey, collection, and conservation of naturally existing chemotypes of medicinal and aromatic plants, considering their potential use in aroma and pharmaceutical industry.

Calcareous sponges of the Western Indian Ocean and Red Sea.

Past taxonomic studies of Western Indian Ocean and Red Sea Calcarea have been few and sporadic (e.g. Schuffner 1877, Jenkin 1908, Row 1909, Dendy 1913, 1916, Voigt et al. 2017, 2018). Nevertheless, approximately 70 species are known from these studies for the considered region, but the descriptions of the older records often lack sufficient details for reliable identification. We studied the Western Indian Ocean Calcarea collection kept in the Naturalis Biodiversity Center. Available specimens numbered 145, collected in the Red Sea, Seychelles, Maldives, Mayotte and Rodrigues, in addition to incidental samples from Oman, the Lakshadweep Islands, the Mozambique Channel, and Eastern South Africa. Using a combination of techniques (in situ and 'on deck' photography, detailed field notes, light microscopic studies and measurements, SEM microscopy, and selected DNA sequencing) we identified 45 species, divided over the two main classes Calcinea (24 spp.) and Calcaronea (21 spp.). Not all species could be definitely assigned to an already described or a new species, as seven remained qualified as 'spec.' or 'aff.' for reasons of insufficient material or lack of details of in situ habitus. Sixteen species appeared to be new to science: Borojevia voigti sp.nov., Borojevia tubulata sp.nov., Borojevia pirella sp.nov., Clathrina rodriguesensis sp.nov., Clathrina maremeccae sp.nov., Clathrina repens sp.nov., Leucascus schleyeri sp.nov., Leucetta sulcata sp.nov., Ute insulagemmae sp.nov., Leucandra pilula sp.nov., Leucandra mozambiquensis sp.nov., Grantessa woerheidei sp.nov., Sycettusa hirsutissima sp.nov., Vosmaeropsis glebula sp.nov., Paraleucilla erpenbecki sp.nov., and Kebira tetractinifera sp.nov. For a selection of the identified species from the Western Indian Ocean and the Red Sea (30 spp.), as well as from Indonesian material (22 spp.) published previously (see Van Soest De Voogd 2015) we obtained sequences of the partial 28S gene of nuclear rDNA (C2-D2 region, cf. Voigt Wörheide 2016). The sequences of the Western Indian Ocean and Red Sea species were used to assign these to genera and families based on a phylogenetic analysis using MEGA pack vs. 06.6 for Mac of the available dataset. The Indonesian sequences supplemented by partial 28S sequences taken from the Sponge Barcode Project website and the NCBI website were included in the phylogenetic analysis to confirm the assignments. The results were compared and discussed with additional information on regional Calcarea not represented in our material. The latter chapter yielded the discovery of a preoccupied name leading to Sycon oscari nom.nov. for a species described from Mauritius.

Morphological, molecular and toxigenic characteristics of Namibian Pseudo-nitzschia species – including Pseudo-nitzschia bucculenta sp. nov.

A field study was undertaken to investigate the occurrence and toxin production of species in the diatom genus Pseudo-nitzschia in Namibian waters, in the extremely productive Benguela upwelling system. From surveys conducted on the R/V Mirabilis and the R/V !Anichab, 52 strains were morphologically determined to species level, supported by nuclear ITS rDNA data. Seven species were identified; P. australis, P. decipiens, P. dolorosa, P. fraudulenta, P. plurisecta, P. pungens var. cingulata, and the new species P. bucculenta F. Gai, C. K. Hedemand, N. Lundholm & Ø. Moestrup sp. nov.Molecular and morphological diversity of the Namibian Pseudo-nitzschia species is discussed. Most importantly, P. bucculenta is both morphologically and phylogenetically most similar to P. dolorosa differing mainly in valve width and densities of striae, poroids and band striae as well as by four hemi-compensatory base changes in the ITS2. Morphological and molecular differences among the strains of P. decipiens suggest a temperate and a warm water subdivision. The geographical and toxigenic characteristics of the identified Pseudo-nitzschia species are described and compared to previous studies. Initial tests of toxin production in all seven species revealed production of domoic acid (DA) in two species: one strain of P. australis (0.074 pg DA cell−1) and two strains of P. plurisecta (0.338 pg DA cell−1 and 0.385 pg DA cell−1).

Genetic examination of the type specimen of Ulva australis suggests that it was introduced to Australia

SUMMARY‘Ana‐aosa’, one of the most common marine green algae in Japan, was described as Ulva pertusa Kjellman in 1897 from Hakodate in northern Japan. Ulva pertusa was considered to be a temperate species, with its native distributional range restricted to northeastern Asia. Although this species has been reported from various regions outside northeastern Asia, these records have been explained as non‐indigenous populations. Recently, on the basis of genetic data and nomenclatural priority, U. pertusa was synonymized with U. australis Areschoug, a species described in 1851 from specimens collected in South Australia. Based on genetic studies, Australian populations identified as U. pertusa had been considered to have originated from Japan. However, the published genetic data on U. australis in Australia have been based only on recent collections and no historical specimens have been examined. We tested the hypothesis that native (true) U. australis is an independent species of very similar morphology to U. pertusa, but that its natural domination of shoreline habitats has been suppressed by introduced populations of U. pertusa from Asia. In the present study, we extracted DNA from the type specimen of U. australis housed in the Swedish Museum of Natural History (S) and obtained DNA sequences of the chloroplast rbcL gene and the nuclear rDNA ITS2 region. Our results show that U. australis and U. pertusa are genetically virtually identical, confirming that U. pertusa is a synonym of U. australis. This suggests that the introduction of U. australis to Australia occurred by the middle of the 19th century, when the type was collected and before there was a direct shipping route between Japan and Australia. We speculate that the introduction of U. australis to Australia occurred as a secondary introduction from non‐indigenous populations in northeastern Asia, but not directly from Japan.

Convoluted history and confusing morphology: Molecular phylogenetic analysis of dicrocoeliids reveals true systematic position of the Anenterotrematidae Yamaguti, 1958 (Platyhelminthes, Digenea)

The Dicrocoeliidae is a highly diverse family of digeneans parasitic in amniotic tetrapods. Detailed molecular phylogenetic analysis of dicrocoeliids is lacking and only a few dicrocoeliids from mammals have been included in previous studies. Sequence data were previously absent for the Anenterotrematidae that shares several morphological characteristics with dicrocoeliids. We examined phylogenetic affinities of several newly sequenced (nuclear 28S rDNA) taxa of dicrocoeliids and anenterotrematids collected from small mammals in Ecuador, Panama, Peru, USA and Vietnam. Our analyses demonstrated that the two anenterotrematid genera (Anenterotrema, Apharyngotrema) belong to the Dicrocoeliidae, placing the Anenterotrematidae into synonymy with the Dicrocoeliidae. Molecular data combined with morphological examination of type and new specimens provided evidence that Parametadelphis and Apharyngotrema are junior synonyms of Metadelphis, with all Metadelphis species lacking a digestive system. Phylogenetic analysis demonstrates that reduction of the alimentary tract in Lutztrema and its loss in Anenterotrema and Metadelphis represent at least two independent evolutionary events. Genera Brachylecithum, Brachydistomum, and Lyperosomum proved to be non-monophyletic, each likely representing more than a single genus. Furthermore, phylogenetic analysis did not support monophyly of the two largest subfamilies of the Dicrocoeliidae (Dicrocoeliinae and Leipertrematinae) with the other two subfamilies not included in this study. Therefore, we propose to abandon the current subfamily division of the Dicrocoeliidae. Analysis of host associations indicates multiple host-switching events throughout evolution of dicrocoeliids. Lastly, analysis of dicrocoeliid geographic distribution revealed that nearly all major clades included taxa from more than a single zoogeographic realm with the exception of the clade Anenterotrema + Metadelphis, found only in the Neotropics.

Genetic diversity studies in endangered desert teak [Tecomella undulata (Sm) Seem] using arbitrary (RAPD), semi-arbitrary (ISSR) and sequence based (nuclear rDNA) markers

Key message Substantial genetic diversity exists in the natural populations of desert teak which needs to be conserved in-situ as well as ex-situ to ensure sustainable utilization and survival of this endangered tree.

Systematics of araphid diatoms with asymmetric rimoportulae or densely packed virgae, with particular attention to Hyalosynedra (Ulnariaceae, Bacillariophyta)

Synedroid diatoms are morphologically characterized by a putative synapomorphic structure called an ocellulimbus, a terminal pore field whose surface is depressed relative to the rest of the valve surface. Among these diatoms is the genus Hyalosynedra Williams and Round. Characters associated with that genus are bilayered wall construction, densely packed virgae and an asymmetric rimoportula. However, these characters diagnose partially overlapping (incongruent) groups. The apical pore field itself, when present, is sometimes sunken deeply, sometimes weakly and sometimes not at all among these diatoms. Here we make additional morphological observations and add molecular sequence data (nuclear SSU rDNA, rbcL, psbC) to attempt to resolve these apparent conflicts. The molecular phylogeny does not resolve any morphological character without homoplasy, but conversely does resolve taxa which share an asymmetric rimoportula and/or densely packed virgae into a single clade we call the “hyalosynedroid” taxa. This clade contains our representatives of Thalassionematales which had been linked to Hyalosynedra by the bilayered wall structure and the possession by some species of an asymmetric rimoportula. We describe three new genera, and a total of 11 new species. Whereas four species of Hyalosynedra were previously described, we describe three new species of Hyalosynedra, describe the new genus Divergita with three species (one new, one transferred from Synedra and one from Hyalosynedra), and the new genus Stricosus with seven new species. A key to species of Hyalosynedra, Divergita and Stricosus is included.

Integrative taxonomy of freshwater ostracodes (Crustacea: Ostracoda) of the Yucatán Peninsula, implications for paleoenvironmental reconstructions in the northern Neotropical region

A three-step integrative taxonomy approach combining molecular, morphological and ecological methods was successfully used to delimit species boundaries within three freshwater ostracode taxa of the Yucatán Peninsula (Mexico): Darwinulidae, Cypretta and Cypridopsis. These species groups were selected because they are well-suited paleobioindicators for late Pleistocene-early Holocene climatic reconstructions in the region, but their high inter-population morphological variability in recent communities represents a challenge for accurate transfer of information to fossil assemblages.As a first step, we analyzed the congruence of two genes of different inheritance, the mitochondrial COI (cytochrome c-oxidase subunit 1), and the nuclear 18S rDNA (small-subunit rDNA). Second, we tested the lineages discriminated for concordant differentiation in morphology, using morphometry of the carapace and typological analysis of the appendages. The third step assessed the association of occurrences and abundances of lineages to environmental variables. The integration of these methods revealed six hidden species within clades previously considered as a single widely distributed nominal species. Of these, Cypretta elongata sp. nov. and Alicenula yucatanensis sp. nov. are herein formally named and described given their morphological distinctness. Three species of Cypridopsis: Cypridopsis sp. [Ca1 ECO-CH-Z-09396], Cypridopsis sp. [Ca2 ECO-CH-Z-09398], Cypridopsis sp. [Ca3 ECO-CH-Z-09401] and a species that may represent a new genus, Cyprididae sp. [Ca1 ECO-CH-Z-09402], are allocated to a confirmed candidate full species status. Cypretta maya and Cypretta sp., considered a priori as separate species, are recognized as a single species with phenotypic plasticity, attributed to its broad ecological tolerances. For paleoenvironmental reconstructions these results have relevance because the particular carapace morphology of the new species can be correlated with specific ecological traits, thus constituting a potential tool for refining interpretations of fossil assemblages.

First endemic freshwater Gammarus from Crete and its evolutionary history-an integrative taxonomy approach.

The Mediterranean islands are known as natural laboratories of evolution with a high level of endemic biodiversity. However, most biodiversity assessments have focused mainly on terrestrial and marine fauna, leaving the freshwater animals aside. Crete is one of the largest islands in the Mediterranean Basin, with a long history of isolation from the continental mainland. Gammarid amphipods are often dominant in macrozoobenthic communities in European inland waters. They are widely used in biomonitoring and exotoxicological studies. Herein, we describe Gammarus plaitisi sp. nov., endemic to Cretan streams, based on morphological characters and a set of molecular species delimitation methods using mitochondrial cytochrome oxidase subunit I and 16S rRNA genes as well as nuclear 28S rDNA, ITS1 and EF1-alpha genes. The divergence of the new species is strongly connected with the geological history of the island supporting its continental origin.

Resolving the Lophiostoma bipolare complex: Generic delimitations within Lophiostomataceae.

Lophiostoma bipolare was taxonomically revised based on the morphological observations and phylogenetic analyses of molecular data from nuclear rDNA SSU-ITS-LSU, TUB, tef1, and rpb2 genes. Twenty-nine strains were morphologically similar to Lo. bipolare. A total of 174 sequences were generated from the Lo. bipolare complex. Phylogenetic analyses based on TUB sequence revealed 11 distinct species within the Lo. bipolare complex. Morphological features of the ascospores and the anatomical structure of the ascomata from both field collections as well as axenic culture, which have been reported previously as variable features at intraspecific levels, were compared to evaluate the taxonomic reliability of these features. To clarify the generic position of the 11 species, phylogenetic analyses were done on SSU-ITS-LSU-tef1-rpb2 gene sequences. The Lo. bipolare complex shared phylogenetic relationships with Pseudolophiostoma and Vaginatispora, and formed an additional five distinct clades from other members of Lophiostomataceae. According to its phylogenetic position, Lo. bipolare sensu stricto was distantly related to Lophiostoma s. str., and formed an independent clade within Lophiostomataceae. Lophiostoma bipolare s. str. could be distinguished from the other lophiostomataceous genera by the clypeus around the ostiolar neck and by the thin and uniformly thick peridium. A novel genus described as Lentistoma was established to accommodate this species, and the epitypification of Lentistoma bipolare (basionym: Massarina bipolaris) was proposed. Other lineages of the Lo. bipolare complex could not be separated on the basis of the ascospore size and sheath variations, but were distinguished based on ascomatal features, such as the existence of the clypeus, brown hyphae surrounding the peridium, and the contexture of the peridium, which were stable indicators of generic boundaries in Lophiostomataceae. Four additional new genera with five new species were recognised based on these morphological differences: Crassiclypeus (C. aquaticus), Flabellascoma (F. cycadicola and F. minimum), Leptoparies (Lep. palmarum), and Pseudopaucispora (Pseudop. brunneospora). Three new species were added to Pseudolophiostoma (Pseudol. cornisporum, Pseudol. obtusisporum, and Pseudol. tropicum) and two new species were added to Vaginatispora (V. amygdali and V. scabrispora). The re-evaluation of the validity of several previously recognised genera resulted in the introduction of two new genera with new combinations for Lophiostoma pseudoarmatisporum as Parapaucispora pseudoarmatispora and Vaginatispora fuckelii as Neovaginatispora fuckelii.

The High Divergence of Two Morphologically Similar Trematode Species of the Genus Nanophyetus of Salmonids from the Data of mtDNA nad1 Gene Sequences

Intestinal flukes of fish (mainly salmonids) belonging to the genus Nanophyetus (Trematoda: Troglotrematidae) are the causative agents of nanophyetiasis, a zoonotic disease of animals and humans, which is widespread in countries in the northern Pacific. Two geographical forms, one from North America and the other from the eastern Eurasia were described within this genus; however, their taxonomic status was debatable. A multilocus analysis of nuclear rDNA sequences applied in this study has shown that these forms are independent nominal species: Nanophyetus salmincola and Nanophyetus schikhobalowi. This study, based on sequencing the mtDNA nad1 gene, has evaluated the genetic variability of N. schikhobalowi from eastern Eurasia (Russia) and compared our data with the results obtained for N. salmincola from North America (United States). The genetic differentiation within the Eurasian sample was 1.4%, that for the North American sample was 0.8%, and differentiation between the samples was 15.5%. High values of genetic divergence and completed sorting of mitochondrial haplotypes confirmed the species independence of the compared geographic forms of Nanophyetus.

First insights into the phylogeny of deep-sea glass sponges (Hexactinellida) from polymetallic nodule fields in the Clarion-Clipperton Fracture Zone (CCFZ), northeastern Pacific

Glass sponges represent a dominant group of megabenthic deep-sea fauna and play a key role in benthic deep-sea ecosystems. Especially in the Clarion-Clipperton Fracture Zone (CCFZ), a potential deep-sea mining area, they grow on polymetallic nodules or on the surrounding sediment. We investigate hexactinellids from the CCFZ to understand the ecological aspects of deep-sea mining and support the development of future pre-mining risk assessments and monitoring actions. Therefore, this study is published as part of a series of studies, all focusing on deep-sea glass sponges from the CCFZ. Resolving genetic relationships between species is still a fundamental as well as challenging task. Especially understudied groups mostly lack resolution. Combining results derived from taxonomic and phylogenetic data gives deeper insights into glass sponge relationships. Here, we present (1) a set of new primers for sequencing mitochondrial 16S rDNA as well as nuclear 18S and 28S rDNA of glass sponges, (2) first DNA sequencing data for 6 hexactinellid genera and 19 species, as well as (3) the most comprehensive phylogenetic tree of hexactinellid sponges to date including data available from previous studies.