Nuclear rDNA Research Articles

Detection and Molecular Characterization of Potentially Pathogenic Free-Living Amoebae from Recreational and Public Soils in Mazandaran, Northern Iran

Background:Free-living amoeba (FLA) belonging to Acanthamoeba spp., Naegleria, and Balamuthia mandrillaris are the soil-born protozoa. This study aimed to survey the occurrence of FLA, including Acanthamoeba spp., B. mandrillaris, Vermamoeba spp., and Naegleria spp., in soil samples collected from various districts of Mazandaran Province (Northern Iran) from July to December 2018.Methods:Overall, 118 soil samples from the recreational and public places were surveyed for the existence of Acanthamoeba spp., Vermamoeba, Naegleria, and B. mandrillaris using both morphological key and molecular tools with genus-specific primers of JDP1, NA, ITS1, and Bal, respectively. To verify the taxonomic status of isolated amoeba, the phylogenetic tree was made based on sequences of 18S rRNA by MEGA (5.05) software with the maximum likelihood model.Results:Overall, 61/118 samples (51.6%) were contaminated with FLA, and based on the sequencing data, 29 isolates were successfully sequenced. Among the samples, all isolated Acanthamoeba (52.4%) belonged to the T4 genotype with amplification of the DF3 region (18S rRNA gene). Internal transcribed spacer (ITS) sequencing revealed the presence of one strain of Naegleria americana. Twenty-eight V. vermiformis were also confirmed based on Nuclear SSU rDNA. Morphological survey and PCR assay did not show any positive samples for B. mandrillaris.Conclusion:The present study indicates the occurrence of FLA in soil sources of the recreational and public places in Mazandaran province that it can be a severe risk to human health. Thus, more studies are expected to survey the infection source in patients with FLA-related diseases.

Phylogeny and revision of the ‘Cubitermes complex’ termites (Termitidae: Cubitermitinae)

AbstractThe phylogeny of the genus Cubitermes Wasmann was reconstructed using two mitochondrial genes (COI and COII) and a fragment of the nuclear 28S rDNA, including samples of Apilitermes Holmgren, Crenetermes Silvestri, Megagnathotermes Silvestri and Thoracotermes Wasmann. Based on our analyses, we recovered these genera within a paraphyletic Cubitermes clade. Cubitermes species are distributed between five main clades, highly reflective of their enteric valve armatures (EVAs). Consequently, Cubitermes is here divided into five monophyletic genera based on phylogenetic analyses and EVAs: Cubitermes (sensu stricto), Isognathotermes Sjöstedt, Nitiditermes Emerson, Polyspathotermes Josens & Deligne, gen.n., and Ternicubitermes Josens & Deligne, gen.n. Moreover, the two species of Megagnathotermes included in this study, exhibiting different EVAs, are phylogenetically distant: M. katangensis Sjöstedt belongs in the genus Polyspathotermes, while M. notandus Silvestri remains as a monotypic genus. During the evolution of the Cubitermes complex, sclerotized EVA spatulae appeared twice independently: three or six spatulae in Polyspathotermes (sulcifrons and oblectatus patterns) and two jaw‐like spatulae in Nitiditermes (sankurensis pattern), which is absent in some species (oculatus pattern). Zoobank registration: http://zoobank.org/urn:lsid:zoobank.org:pub:576AAFF8‐63C6‐4962‐A8F5‐75C9317AEA7B.

Molecular characterization of enteric coccidia from domestic ferrets (Mustela putorius furo).

Combined morphometric and molecular characterization of coccidia that infect domestic ferrets (Mustela putorius furo) was completed to improve the diagnostic specificity of 'coccidiosis' in this host. Coccidia-positive fecal samples (n=11) and formalin-fixed paraffin-embedded intestinal tissues (n=3) from domestic ferrets were collected from diagnostic laboratories in Canada and Europe. An average of 3.5 and 13 domestic ferret fecal samples per year were coccidia-positive when tested by Canadian and European diagnostic laboratories, respectively, during the period 2008-2015. Oocyst morphometrics and sequence genotyping at two loci (nuclear 18S rDNA [nu 18S] and mitochondrial cytochrome c oxidase subunit I [mt COI]) were conducted on all samples. The first nu 18S and mt COI sequences for Cystoisospora laidlawi, and the first mt COI sequence for Eimeria furonis were generated during this study. Phylogenetic analysis of the mitochondrial COI sequences demonstrated that E. furonis was most closely related to E. ictidea isolated from a black-footed ferret (Mustela nigripes) and that C. laidlawi was closely related to C. canis and C. felis. The identifications provided by diagnostic laboratories of the specific parasite species present in a sample showed poor agreement with their identifications based on genotyping obtained in this study. Molecular techniques appear to be essential for determining the specific coccidial species responsible for individual and group outbreaks of coccidiosis and for further understanding of eimeriid host-parasite relationships.

Phlebopus roseus, a new edible bolete from China, is associated with insects and plants

ABSTRACT Phlebopus roseus is described as new based on collections from southwest China. Phylogenetic analyses of nuclear rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and portions of nuclear 28S rDNA (28S), translation elongation factor 1-alpha (tef1), and the largest and second largest subunits of RNA polymerase II (rpb1, rpb2) support P. roseus as a novel species in the genus Phlebopus (Boletinellaceae, Boletales). The new species resembles P. portentosus but differs from it in that mature basidiomata have a bright rose-red-colored stipe and a radiate tubular hymenophore with nested pores. Despite extensive searching, P. roseus has only been found at four sites within a 24-hectare orchard dominated by Eriobotrya japonica, which is agriculturally important given its fruit production (loquats). Therefore, this species appears to be endemic and geographically restricted. The ecology of this bolete is also unique. In line with the trophic behavior of other species in the Boletinellaceae, our observations indicate that P. roseus forms a symbiotic association with the scale insect Coccus hesperidum, identified through sequence analysis of its mitochondrial cytochrome c oxidase subunit I (COI) region, to form fungus-insect galls that develop on roots of E. japonica trees. Phlebopus roseus is an edible mushroom species and is collected from the type location by farmers and sold commercially in limited quantities at local markets alongside P. portentosus and other fungi.

Host-induced phenotypic plasticity in Saccocoelioides lamothei Aguirre-Macedo and Violante-González, 2008 (Digenea: Haploporidae) a parasite of freshwater, brackish and marine fishes from Middle America.

Saccocoelioides is a genus of trematodes associated with fishes from the Americas. In the current research, morphologically distinct specimens of Saccocoelioides spp. were collected from six countries in Middle America. Specimens were sequenced using three molecular markers, the domains D1-D3 of the large subunit (LSU) from the nuclear rDNA, the cytochrome c oxidase subunit 1 (cox1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) from mitochondrial DNA. A total of 74 new sequences were compared and aligned with other sequences available in GenBank. Maximum likelihood and Bayesian inference analyses were inferred from the LSU and cox1 datasets, revealing unequivocally that all the specimens correspond to S. lamothei. A haplotype network was built with 119 sequences of the nad1 gene. The network detected 57 distinct haplotypes divided into three haplogroups. To explore morphological differences among samples of S. lamothei, 17 morphological features were measured from 53 specimens from three fish families: Eleotridae, Mugilidae and Gobiidae. Principal component analysis yielded three main polygons that corresponded with each family analysed, suggesting host-induced phenotypic plasticity. The current evidence suggests that S. lamothei infects at least five fish families along the Pacific coasts of Mexico, Guatemala, El Salvador, Honduras, Nicaragua and Costa Rica.

Exposing the Barcoding Void: An Integrative Approach to Study Snail-Borne Parasites in a One Health Context.

Trematodes are snail-borne parasites of major zoonotic importance that infect millions of people and animals worldwide and frequently hybridize with closely related species. Therefore, it is desirable to study trematodiases in a One Health framework, where human and animal trematodes are considered equally important. It is within this framework that we set out to study the snail and trematode communities in four artificial lakes and an abattoir in Zimbabwe. Trematode infections in snails were detected through multiplex PCR protocols. Subsequently, we identified snails by sequencing a partial mitochondrial cytochrome c oxidase subunit I (COI) fragment, and trematodes (adults from the abattoir and larval stages detected in snails) using COI and nuclear rDNA markers. Of the 1,674 collected snails, 699 were molecularly analyzed, in which we identified 12 snail and 19 trematode species. Additionally, three parasite species were sampled from the abattoir. Merely four trematode species were identified to species level through COI-based barcoding. Moreover, identification of members of the superfamilies Opisthorchioidea and Plagiorchioidea required a phylogenetic inference using the highly conserved 18S rDNA marker, as no related COI reference sequences were present in public databases. These barcoding challenges demonstrate a severe barcoding void in the available databases, which can be attributed to the neglected status of trematodiases. Adding to this, many available sequences cannot be used as different studies use different markers. To fill this gap, more studies on African trematodes, using a standardized COI barcoding region, are desperately needed.

Monadelpha (Euphorbiaceae, Plukenetieae), a new genus of Tragiinae from the Amazon rainforest of Venezuela and Brazil.

Monadelpha L.J.Gillespie & Card.-McTeag., gen. nov., is described as a new member of Euphorbiaceae tribe Plukenetieae subtribe Tragiinae, to accommodate Tragiaguayanensis, a species known from western Amazonas, Venezuela and, newly reported here, from Amazonas, Brazil. The genus is unique in the subtribe for having 5-colpate pollen and staminate flowers with filaments entirely connate into an elongate, cylindrical staminal column terminated by a tight cluster of anthers. Phylogenetic analyses based on nuclear rDNA ITS and sampling 156 accessions across the diversity of Tragiinae (all 12 genera and 77 of ~195 species) also support Monadelpha as a distinct lineage that is separate from Tragia. A revised key to the genera of Tragiinae in South America and Central America is provided.

Integrative systematics reveals the new land-snail genus Taphrenalla (Eupulmonata: Ariophantidae) with a description of nine new species from Thailand

Abstract The endemic terrestrial Crown Snail from Southern Thailand formerly in Macrochlamys Gray, 1847 is now described as Taphrenalla Pholyotha & Panha gen. nov., based on comparative morphology and molecular data. Overall, eleven Taphrenalla species are now recognized, including two nominal species: T. asamurai and T. diadema. A total of nine new species are proposed: T. alba sp. nov., T. conformis sp. nov., T. corona sp. nov., T. dalli sp. nov., T. incilis sp. nov., T. macrosulcata sp. nov., T. parversa sp. nov., T. pygmaea sp. nov. and T. zemia sp. nov. The molecular phylogeny constructed from the mitochondrial cytochrome oxidase c subunit I (COI) and 16S rRNA gene fragments plus the nuclear 28S rDNA gene fragments revealed that Taphrenalla gen. nov. is monophyletic with a well-supported clade. The diagnostic characters of Taphrenalla gen. nov. are the shell sculpture with several radial grooves, body with well-developed colourful stripes running from the head to tail, and genitalia similar to Macrochamys but with an un-coiling epiphallic caecum. The spermatophore has one or two spines near the sperm sac and a spineless tail.

Two new enigmatic species of Rhinebothrium (Cestoda: Rhinebothriidae) from the Persian Gulf: notes on generic traits and host specificity

In the course of a three-year study conducted from January 2017 till November 2019, a total of 279 individuals belonging to 11 batoid species off northern coasts of the Persian Gulf and the Gulf of Oman were examined for the genus Rhinebothrium Linton, 1890 (Rhinebothriidea: Rhinebothriidae). Infection of Maculabatis randalli (Last, Manjaji-Matsumoto & Moore, 2012), M. arabica Manjaji-Matsumoto & Last et al., 2016, and Pateobatis fai (Jordan & Seale, 1906) with rhinebothriid cestodes is reported for the first time. In total, seven rhinebothriid species were recognized. Rhinebothrium klimpeli sp. nov. from Pateobatis fai and Rhi. atabaki sp. nov. from Maculabatis randalli are described herein. Both species differ from one another and their congeners on the basis of various characteristics of the scolex and proglottids. In order to create a DNA Barcoding library for local species and perform a phylogenetic analysis, different marker genes for seven species of Rhinebothrium were partially sequenced here for the first time; nuclear small subunit rDNA (SSU) and large subunit rDNA (LSU), and the Cytochrome c oxidase subunit I (COI) gene. Both Bayesian inference and maximum likelihood phylogenetic analyses using nuclear markers confirmed species boundaries recognized by morphological characteristics. Furthermore, both new species are classified into the genus Rhinebothrium based on molecular data. The description of Rhi. klimpeli sp. nov. leads us to modify the diagnosis of the genus Rhinebothrium in introducing the character 'bearing pre- and postvaginal testes' and 'lacking postovarian testes'. Rhinebothrium verticillatum (Subhapradha, 1955) is redescribed herein with designation of the neotype identified from the type host Rhynchobatus laevis (Bloch & Schneider, 1801) deposited at the Collection of the Zoological Museum, University of Tehran (ZUTC). All parasite species except Rhi. verticillatum were found in more than one host species from different families. Among them, Rhi. atabaki sp. nov. infects seven of 11 host species and seems to have the lowest host specificity. Furthermore, some implications for host specificity of rhinebothriid species are discussed. http://zoobank.org/urn:lsid:zoobank.org:pub:1CD0BC57-2741-4352-A7C5-CD6C36BF967C http://zoobank.org/urn:lsid:zoobank.org:act:9AA8D331-7936-49D7-8CA4-0B6BC927D0AC http://zoobank.org/urn:lsid:zoobank.org:act:5B21A19B-89E8-48E9-A0FF-FAB57387912F

New pelagophytes show a novel mode of algal colony development and reveal a perforated theca that may define the class.

Pelagophytes (Heterokonta) are a morphologically diverse class of marine algae historically united only by DNA sequences. We established clonal cultures of sand-dwelling pelagophytes collected from intertidal pools around Australia. Phylogenetic trees based on nuclear 18S rDNA and plastid rbcL, psaA, psaB, psbA, and psbC sequences revealed two new genera, Gazia and Glomerochrysis, related to Aureoumbra in a distinct lineage within the Sarcinochrysidaceae (Pelagophyceae). The three new species (Gazia saundersii, Gazia australica, and Glomerochrysis psammophila), along with an Australian strain of Aureoumbra geitleri, are characterized by dominant benthic stages that differ significantly from one another, while occasionally producing classic heterokont zoospores. The benthic stage of Ga. saundersii has a novel development not observed in any other colonial alga, consisting of large, spherical colonies (up to 140μm in diameter) containing c. 2,500 cells that eventually differentiate and segregate into a large number of daughter colonies that are subsequently liberated. Alternatively, colonies may differentiate into a mass of zoospores that escape and settle to develop into new colonies. In Gl. psammophila, cubic packets of cells form large sticky clusters that bind sand together, while Ga. australica and A. geitleri are unicellular species. Using fixation by high-pressure freezing, a distinctive perforated theca was observed by TEM in all genera of this lineage, and we hypothesize this unique covering may be the first morphological feature to characterize most, if not all, pelagophytes. This study substantiates the diverse nature of sand-dwelling pelagophytes as well as their mechanisms for thriving in a dynamic habitat.

Unexpected morphological and molecular diversity of trematode (Haploporidae: Forticulcitinae) parasites of mullets from the ocean Pacific coasts in Middle America.

Two new genera and four new species from subfamily Forticulcitinae are described from the intestines of white mullet (Mugil curema) and flathead grey mullet (Mugil cephalus) collected in 27 localities across a wide geographical range on Pacific Ocean slopes comprising three countries in Middle America: Mexico, Guatemala and Costa Rica. The new genera Ekuarhuni n. gen. and Overstreetoides n. gen. had to be erected to accommodate two new species, Ekuarhuni papillatum n. sp. and Overstreetoides pacificus n. sp., with unique morphological traits that differentiate them from the two genera described previously. In addition, two new species, Forticulcita minuta n. sp. and Forticulcita isabelae n. sp., were described, which were characterized as exhibiting a small body size (< 1100μm long). These new species were classified as the diminutive morphotype, together with three other congeneric species of Forticulcita. Forticulcita minuta n. sp. is distinguished by being the smallest species within the genus (< 305μm). Meanwhile, Forticulcita isabelae n. sp. is distinguished by its body size and testis length. In specimens of the four new species, sequencing was performed with two molecular markers, the large subunit (LSU) and the internal transcribed spacer 2 (ITS2) of nuclear rDNA, and the results were compared with other sequences available in GenBank. Phylogenetic analyses performed with the combined dataset of the two nuclear molecular markers (LSU + ITS2) placed all the analysed species within the clade of Forticulcitinae with strong bootstrap support (100%) and a high Bayesian posterior probability (1.0). The four new species showed differences in abundance in their definitive hosts and were widely distributed along the Pacific Ocean coasts of Mexico, Guatemala and Costa Rica, in Middle America.

Molecular approach for analysis of in situ feeding by the dinoflagellate Noctiluca scintillans

The red tide forming heterotrophic dinoflagellate Noctiluca scintillans is common in temperate to tropical waters around the world. Understanding the in situ prey of N. scintillans is essential for elucidating its role in marine microbial food webs. In this study, we applied two polymerase chain reaction (PCR)-based cloning techniques, a predator-specific restriction enzyme, and a blocking primer. The PCR of nuclear 18S rDNA was performed on single N. scintillans cells that were collected from Ishinomaki Bay, Japan, in May 2018. The maximum detection rates of non-Noctiluca sequences were 56% using the restriction enzyme method and 87% with the blocking primer method, representing a broad taxonomic range of organisms, including diatom, dinoflagellate, bolidophyte, haptophyte, euglenophyte, green algae, golden algae, ciliate, heliozoa, copepod, brown seaweed, sponge, bivalve, and polychaete. The diverse DNA was probably ingested by N. scintillans directly or indirectly through secondary predation or ingestion of marine snow or detritus containing many organisms. The application of molecular approaches to various species may reveal undiscovered interactions within the phytoplankton community, including prey-predator, or symbiotic relationships.

A phylogenetic analysis of Momordica (Cucurbitaceae) in India based on karyo-morphology, nuclear DNA content and rDNA ITS1-5.8S-ITS2 sequences.

The infrageneric delimitation of Momordica, a medicinally important genus of Cucurbitaceae, is ill-defined until date. Momordica chromosomes are extremely small and are difficult to stain and visualize because of the dense cytoplasmic background. We have conducted karyomorphometric analysis by EMA method in five Indian Momordica species, and the nuclear genome sizes were estimated by flow cytometry for the first time. The somatic chromosome numbers ranged from 2n = 18 to 56 in the species. We have resolved previously disputed chromosome numbers in M. cymbalaria and M. dioica as 2n = 18 (lowest) and 2n = 56, respectively. Chromosome counts in the other species were re-confirmed as 2n = 22 in M. charantia, 2n = 28 in M. cochinchinensis and 2n = 56 in M. subangulata. The largest genome size was recorded in M. cymbalaria (3.74pg 2C-1), while the smallest size (0.72pg 2C-1) was detected in M. charantia var. charantia. The nuclear genome sizes were analysed in comparison to chromosome numbers and total chromosome lengths of the species. Karyomorphometric indices showed comparable symmetric karyotypes in the species except in M. cymbalaria having tendency towards asymmetry. The UPGMA phenogram and principle component analysis based on nuclear DNA contents and karyomorphometric parameters demonstrated interspecies differences, intraspecific distinction within M. charantia varieties and highlighted distinction of M. cymbalaria. This study was further supported by the rDNA ITS sequence-based phylogenetic analysis which revealed the monophyletic origin of the Indian members of Momordica and clarified the intraspecies relationship among the studied members. As a whole, the study brought out new insights on species diversification within the genus Momordica in India and would benefit further studies on biosystematics and plant breeding programmes.

Phylogeny of Adramini (Diptera, Tephritidae) based on integrative evidence

AbstractSpecies of the tribe Adramini (Tephritidae: Trypetinae) are usually slender, and some specific species have eyes borne at the ends of their long stalks. This tribe is mainly distributed in the tropics and subtropics of the Afrotropical, Oriental and Australasian Regions; relatively few species occur in the Palearctic and Nearctic Regions. The phylogeny of the tribe Adramini is presented here based on analysis of morphological and molecular information (DNA sequences of nuclear 28S rDNA, mitochondrial COI and COII, and 16S rDNA genes) for its representative species in most genera. Three monophyletic groups (Adrama‐com‐group, Pelmatops‐com‐group and Dimeringophrys‐com‐group) were discovered in the combined morphological and molecular tree. The results showed moderate support for the monophyly of Adramini and strong support for most of its genera. However, Euphranta appears to be polyphyletic. Sapadrama, Celidodacus and Euphranta are basal taxon, and Coelopacidia, Soita and Trypanophion are closely related to the stalk‐eyed fruit flies (Pelmatops + Pseudopelmatops). A hypothesis regarding the morphology–function relationships for two main groups of Adramini (Adrama‐group and Pelmatops‐group) with different evolving probabilities is inferred. Sapadrama is proposed be removed from Adramini; a new genus, Ichneumonomacula Chen gen. n. and a new species Ichneumonmacula wangyongi Chen sp. n., are recognized and described, and a key to recognize the genera of Adramini around the world is provided.

Molecular phylogeny and chromosomal evolution of endemic species of Sri Lankan Anacardiaceae

Family Anacardiaceae comprises 70 genera and approximately 985 species distributed worldwide. Sri Lanka harbours 19 species in seven genera, among these 15 are endemics. This study focuses on regionally restricted endemics and native Anacardiaceae species, which have not been investigated before at molecular and cytological level. Nuclear rDNA ITS and plastid matK regions were sequenced for ten species, having nine endemics and one native, and incorporated into the existing sequence data for phylogenetic analyses. The topologies resulting from maximum parsimony, maximum likelihood and Bayesian inference are congruent. Family Anacardiaceae forms a monophyletic group having monophyletic subfamily Anacardioideae and paraphyletic subfamily Spondioideae. Tribes Anacardieae, Semecarpeae and Rhoeae form subclades within the major clade of Anacardioideae. All the endemic species occupy correct position in the molecular phylogeny as per the existing classification except for Campnosperma zeylanica, which shows a close relationship to members of the subfamily Spondioideae. The position of non-native species Trichoscypha acuminata also remains unclear. Chromosome counts and karyograms were constructed for five endemic species. The chromosome numbers incorporated in the tree range from 2n = 28–58. Species of tribe Rhoeae have the lowest chromosome number (2n = 24, 28, 30) while species of tribe Semecarpeae have the higher numbers (2n = 50, 52, 58). Chromosome numbers mapped on the phylogeny shows that dysploidy had played a role in the evolution of the species of the family Anacardiaceae in Sri Lanka.

Genotyping ITS and matK regions of Hedera nepalensis K. Koch in Vietnam

Genotyping ITS and matK regions of Hedera nepalensis K. Koch in Vietnam

Production of chitosan from Oyster mushroom for α-amylase immobilization

Experiments were carried out to produce chitosan from a locally available mushroom and to use the produced chitosan as a matrix to immobilize α-amylase. On the basis of morphological characteristics and the sequence of the internal transcribed spacer region of the nuclear rDNA, the fungal sample was identified as Pleurotus ostreatus. The average crude chitin content in the dried fruit body of P. ostreatus was 24.11%. The average yield of chitosan from P. ostreatus was 163.3 mg/g dry weight and the degree of deacetylation of the produced chitosan was 73.42%. This chitosan was used as a matrix to immobilize α-amylase. The diameter of the α-amylase immobilized beads ranged from 1839 - 2273 μm. The amount of reducing sugar produced from starch by using free α-amylase and chitosan-immobilized α-amylase was 1.710 and 1.508 mg/ml, respectively. Immobilized enzyme produced only 11.81% less reducing sugar than that of the soluble enzyme in the first cycle. However, immobilized α-amylase was easily recovered from the product and reused for two more cycles which was not possible with the same soluble free enzyme. Considering the total production of reducing sugar in three cycles, chitosan-immobilized α-amylase was found to be more productive and cost-effective than conventional soluble enzymatic reaction.

Re-Evaluation of the Order Sordariales: Delimitation of Lasiosphaeriaceae s. str., and Introduction of the New Families Diplogelasinosporaceae, Naviculisporaceae, and Schizotheciaceae.

The order Sordariales includes the polyphyletic family Lasiosphaeriaceae, which comprises approximately 30 genera characterized by its paraphysate ascomata, asci with apical apparati, and mostly two-celled ascospores, which have a dark apical cell and a hyaline lower cell, frequently ornamented with mucilaginous appendages. To produce a more natural classification of this family, we carried out a phylogenetic analysis based on sequences of the internal transcribed spacer region (ITS), the nuclear rDNA large subunit (LSU), and fragments of ribosomal polymerase II subunit 2 (rpb2) and β-tubulin (tub2) genes of several isolates from soil and of reference strains of the Sordariales. As a result, Lasiosphaeriaceae s. str. has been circumscribed for the clade including the type species of the genus Lasiosphaeria and, consequently, its description emended. In addition, the new families Diplogelasinosporaceae, Naviculisporaceae, and Schizotheciaceae are introduced to accommodate those taxa located far from the Lasiosphaeriaceae s. str. Moreover, we propose the erection of the new genera Areotheca, Lundqvistomyces, Naviculispora, Pseudoechria, Pseudoschizothecium, and Rhypophila based on morphological and sequence data. New combinations for several species of the genera Cladorrhinum, Jugulospora, Podospora, Schizothecium, and Triangularia are proposed, their descriptions are emended, and dichotomous keys are provided to discriminate among their species.

Genetic Evidence for a Mixed Composition of the Genus Myoxocephalus (Cottoidei: Cottidae) Necessitates Generic Realignment.

Sculpin fishes belonging to the family Cottidae represent a large and complex group, inhabiting a wide range of freshwater, brackish-water, and marine environments. Numerous studies based on analysis of their morphology and genetic makeup frequently provided controversial results. In the present work, we sequenced complete mitochondrial (mt) genomes and fragments of nuclear ribosomal DNA (rDNA) of the fourhorn sculpin Myoxocephalus quadricornis and some related cottids to increase the power of phylogenetic and taxonomic analyses of this complex fish group. A comparison of the My. quadricornis mt genomes obtained by us with other complete mt genomes available in GenBank has revealed a surprisingly low divergence (3.06 ± 0.12%) with Megalocottus platycephalus and, at the same time, a significantly higher divergence (7.89 ± 0.16%) with the species of the genus Myoxocephalus. Correspondingly, phylogenetic analyses have shown that My. quadricornis is clustered with Me. platycephalus but not with the Myoxocephalus species. Completely consistent patterns of divergence and tree topologies have been obtained based on nuclear rDNA. Thus, the multi-gene data in the present work indicates obvious contradictions in the relationships between the Myoxocephalus and Megalocottus species studied. An extensive phylogenetic analysis has provided evidence for a closer affinity of My. quadricornis with the species of the genus Megalocottus than with the species of the genus Myoxocephalus. A recombination analysis, along with the additional GenBank data, excludes introgression and/or incorrect taxonomic identification as the possible causative factors responsible for the observed closer affinity between the two species from different genera. The above facts necessitate realignment of the genera Myoxocephalus and Megalocottus. The genetic data supports the two recognized genera, Myoxocephalus and Megalocottus, but suggests changing their compositions through transferring My. quadricornis to the genus Megalocottus. The results of the present study resolve the relationships within a complex group of sculpin fishes and show a promising approach to phylogenetic systematics (as a key organizing principle in biodiversity research) for a better understanding of the taxonomy and evolution of fishes and for supplying relevant information to address various fish biodiversity conservation and management issues.

Molecular Detection for Two Abalone Shell-boring Species Polydora haswelli and P. hoplura (Polychaeta, Spionidae) from Korea using 18S rDNA and cox1 Markers

Polydora species (Polychaeta: Spionidae) are morphologically similar and have shell-boring abilities that are harmful to commercially important mollusks; genetic analysis is therefore important for their identification. Here, molecular evidence for Polydora species in Korean cultured abalone (Haliotis discus hannai) is first reported. DNA barcode markers (Nuclear 18S rDNA and mitochondrial cox1 regions) identified Polydora haswelli and P. hoplura in infested abalone shells. The cox1 marker was efficient in detecting Polydora species even in samples contaminated with shell debris. The findings provide useful information for the accurate identification of Polydora species and for monitoring harmful spionids in mollusk aquaculture farms.