Nuclear Ribosomal RNA Gene Research Articles

The potential use of mitochondrial ribosomal genes (12S and 16S) in DNA barcoding and phylogenetic analysis of trematodes

BackgroundGenetic markers like the nuclear ribosomal RNA (rRNA) genes, internal transcribed spacer regions, mitochondrial protein-coding genes, and genomes have been utilized for molecular identification of parasitic trematodes. However, challenges such as the design of broadly applicable primers for the vast number of species within Digenea and the genetic markers’ ability to provide sufficient species-level resolution limited their utility. This study presented novel and broadly applicable primers using the mitochondrial 12S and 16S rRNA genes for Digenea and aimed to show their suitability as alternative genetic markers for molecular identification of orders Plagiorchiida, Echinostomida, and Strigeida.ResultsOur results revealed that the mitochondrial 12S and 16S rRNA genes are suitable for trematode molecular identification, with sufficient resolution to discriminate closely related species and achieve accurate species identification through phylogenetic placements. Moreover, the robustness of our newly designed primers to amplify medically important parasitic trematodes encompassing three orders was demonstrated through successful amplification. The convenience and applicability of the newly designed primers and adequate genetic variation of the mitochondrial rRNA genes can be useful as complementary markers for trematode molecular-based studies.ConclusionsWe demonstrated that the mitochondrial rRNA genes could be alternative genetic markers robust for trematode molecular identification and potentially helpful for DNA barcoding where our primers can be widely applied across the major Digenea orders. Furthermore, the potential of the mitochondrial rRNA genes for molecular systematics can be explored, enhancing their appeal for trematode molecular-based studies. The novelty of utilizing the mitochondrial rRNA genes and the designed primers in this study can potentially open avenues for species identification, discovery, and systematics in the future.

Redescription of Ptychobothrium belones (Dujardin, 1845) (Cestoda: Bothriocephalidea) from needlefishes (Beloniformes: Belonidae) in the Mediterranean Sea.

The bothriocephalidean tapeworm Ptychobothrium belones (Dujardin, 1845) Lönnberg, 1889 is redescribed on the basis of new materials collected from the intestine of the Mediterranean needlefish Tylosurus imperialis (Rafinesque) (Beloniformes: Belonidae) off Tunisia in the central Mediterranean Sea. This species is characterised by a laterally compressed hexagonal to fan-shaped scolex with an apical disc and two well-developed bothria, an oval ovary and numerous testes arranged in lateral and median fields. The typical diagnostic feature of P. belones is the medullary yolk follicles located between the muscle fibres of the internal longitudinal muscles (paramuscular) and the uterine duct strongly sinuous, S-shaped in mature proglottides. New molecular data (partial sequence of nuclear large subunit ribosomal RNA gene) confirmed the close relationship with specimens previously collected by Brabec et al. (2006, 2015) from the banded needlefish Strongylura leiura (Bleeker) off the Maldives, Indian Ocean, but these specimens are most likely another species. Thus, Ptychobothrium belones as previously reported represents most likely more than one species and its cosmopolitan distribution and host specificity revised by Kuchta et al. (2008b) should be revisited. The type material of P. belones was reported as collected in Mediterranean Sea off Sète, France, from Belone belone (Linnaeus); however, this host was probably misidentified, being confused with morphologically similar small specimens of T. imperialis. This study confirms that P. belones is an exclusive parasite of needlefishes (Belonidae) but at least one further species infecting S. leiura may exist.

First Report of Mucor irregularis Causing Postharvest Fruit Rot on Garcinia mangostana in China.

Mangosteen (Garcinia mangostana Linn.) is an evergreen fruit tree grown in Southeast Asia. The fruit of Mangosteen is dark purple spherical with a diameter of 5 to 8 centimeter, a hard rind and white flesh. The seeds and pericarps of the fruit have long been utilized as a traditional medicine in Southeast Asia (Ovalle-Magallanes et al. 2017). Surveys were conducted during the months from March to April 2019 of fruit markets located in Wujing Town, Shanghai (31°02'33″ N, 121°28'04″ E), one noteworthy postharvest disease was observed. Overall 116 fruit samples were collected from 5 different supermarkets, 10.3% (12) of the fruits were found infected with the same symptoms during cold storage. The infected fruits were subjected to pathogen isolation and characterization. The junction areas between diseased and healthy tissues were cut into small pieces of diameter 5 mm, which were surface sterilized with 0.5% NaClO solution for 3 min, followed by rinsing in sterile water thrice, and then the tissues were placed on sterilized potato dextrose agar (PDA) plates supplemented with 50 μg/ml streptomycin. The colony edges of resulted fungal cultures were cut and removed to new PDA plates for purification after 2 days incubation at 28°C. After 3 rounds of subculture, a total of 19 purified isolates were obtained and these isolates showed similar phenotypes in morphological characters. The fungal colonies were fast growing (25 mm per day at 28°C on PDA plates) and were circular shaped with massive aerial hyphae, white to ochraceous color. Non-septate hyphae were hyaline and the septa were found only at the branching points. Sporangiophores arose from hyphae or from stolons. Sporangiospores were hyaline, smooth-walled, mostly subspherical to ellipsoidal, and measured 1.34 to 2.73 µm (width) and 3.12 to 6.19 µm (length) (1.98 ± 0.36×4.46 ± 0.69, n=100)(Alvarez et al. 2011). Three independent isolates (GM-07, GM-16, GM-19) were selected used for further genetic analysis and pathogenicity test. The internal transcribed spacer (ITS) and the D1/D2 region of the nuclear large subunit ribosomal RNA gene (LSU) were amplified by primers ITS1/ITS4 and primers NL1/LR3, respectively (Walther et al. 2013). BLASTn analyses of ITS (MN081612, MN081613, MN087659) and LUS (MN080423, MN080422, MN080432) sequences exhibited 99 to 100% identify to Mucor irregularis strains HZ640, extr25 and CBS977.68 (Accession numbers KC461926, MH510278 of ITS and JX976214 of LUS). These observed morphological features and molecular identification indicated that the pathogen possessed identical characteristics as Mucor irregularis Stchigel, Cano, Guarro & Ed. Alvarez (Alvarez et al. 2011). To conduct a pathogenicity test (Al-Sadi et al. 2014), 20 µl of spore suspension (106 spores ml-1) were inoculated into five healthy mangosteen fruits by sterile syringes, meanwhile another five healthy fruits were injected with sterilized water as controls. All inoculated fruits were kept in sealed plastic boxes at 28°C with a 12L/12D illumination cycle and 90% relative humidity. After about 3 to 7 days, the inoculated materials showed similar symptoms to those on the original diseased fruits, while control samples remained symptomless. M. irregularis was successfully re-isolated from symptomatic fruits, fulfilling Koch's postulates. Only three plant species that can be infected by M. irregularis have been reported so far, including key lime from Oman (Al-Sadi et al. 2014), corn from China (Peng et al. 2015), and sorghum from Brazil (Lima et al. 2018). To our knowledge, this is the first report of M. irregularis causing the postharvest rot on mangosteen fruit in China.

Species diversity, molecular phylogeny and ecological habits of Cyanosporus (Polyporales, Basidiomycota) with an emphasis on Chinese collections.

Cyanosporus is a genus widely distributed in Asia, Europe, North America, South America and Oceania. It grows on different angiosperm and gymnosperm trees and can cause brown rot of wood. Blue-tinted basidiomata of Cyanosporus makes it easy to distinguish from other genera, but the similar morphological characters make it difficult to identify species within the genus. Phylogeny and taxonomy of Cyanosporus were carried out based on worldwide samples with an emphasis on Chinese collections, and the species diversity of the genus is updated. Four new species, C.flavus, C.rigidus, C.subungulatus and C.tenuicontextus, are described based on the evidence of morphological characters, distribution areas, host trees and molecular phylogenetic analyses inferred from the internal transcribed spacer (ITS) regions, the large subunit of nuclear ribosomal RNA gene (nLSU), the small subunit of nuclear ribosomal RNA gene (nSSU), the small subunit of mitochondrial rRNA gene (mtSSU), the largest subunit of RNA polymerase II (RPB1), the second largest subunit of RNA polymerase II (RPB2), and the translation elongation factor 1-α gene (TEF). Our study expanded the number of Cyanosporus species to 35 around the world including 23 species from China. Detailed descriptions of the four new species and the geographical locations of the Cyanosporus species in China are provided.

Short communication: Algal genotypes in White Syndrome infected coral Acropora muricata from the Karimunjawa Islands, Indonesia

Abstract. Wijayanti DP, Nadia M, Indrayanti E, Haryanti D, Bachtiar M. 2021. Short communication: Algal genotypes in White Syndrome infected coral Acropora muricata from the Karimunjawa Islands, Indonesia. Biodiversitas 23: 415-423. The physiological equilibrium of the coral holobiont may help the corals to survive under various environmental threats. The symbiont genotype is thought to play an important role in the host's susceptibility to thermal stress and coral disease incidence. We identified the Symbiodiniaceae diversity of coral Acropora muricata infected by White Syndrome (WS). Samples were collected from Cemara Kecil Island and Barakuda Beach, Kemujan Island, Karimunjawa Islands. We applied DNA barcoding of 28S nuclear ribosomal RNA gene of the healthy colonies, healthy tissue, and lesions of diseased colonies to perform their symbiont type identification. There were two different symbiont types observed from the phylogenetic analysis, which formed two clades. A clade consisting all Cemara Kecil Island samples while the other with all Barakuda Beach samples except for one. All samples harbored symbiont types from the genus Cladocopium. The members of the genus were reported to have various physiological properties which may support the host resilience when facing the disease. A robust study is needed to reveal the influence of symbiont genotypes on the occurrence of coral disease and the susceptibility of the coral host.

Taxonomy and Phylogeny of a new species of Pseudocercospora on Solanum nigrum from India

A new species Pseudocercospora solanicola is discovered causing severe leaf spot disease on Solanum nigrum from Sonebhadra. Species is described and illustrated based on morphology, molecular sequence analysis and phylogeny of the large subunit (LSU) and internal transcribed spacer region of the nuclear ribosomal RNA (rRNA) gene. Morphologically, new species has similarity with allied species in Pseudocercospora but differs in having black circular, velvety leaf spot and size of stromata, conidia and conidiophores. A key to Pseudocercospora species on Solanum has been provided.

Paraboeremia yungensis sp. nov., a new fungal species isolated from Las Yungas, South America, with promising tyrosinase production potential

In the context of a bioprospection programme for tyrosinase/L-DOPA- and melanin-producing fungal strains for biotechnological purposes, a hyperproducer isolate was obtained from Las Yungas rainforest, a relevant biodiverse ecoregion in North-Western Argentina. The selected strain was preliminarily identified as Paraboeremia sp. This is, to the best of our knowledge, the first native reported species of this genus in South America. Single-gene and multi-locus analyses of the internal transcribed spacer nuclear ribosomal RNA gene region (ITS), partial large subunit 28S nrDNA region (LSU), RNA polymerase II region (RPB2) and partial β-tubulin gene (TUB2) alignments were carried out to define the phylogenetic identity of this strain. As part of a polyphasic identification approach, these results were combined with morphological studies of active cultures growing on malt extract, oatmeal and potato dextrose agar plates. Incubation was performed under diverse conditions to stimulate sporulation for the subsequent micromorphological analysis. Microphotographs of pycnidia and conidia were taken with a scanning electron microscope. Maximum likelihood and Bayesian Inference analyses supported the location of the strain within the genus Paraboeremia, whilst morphological features allowed distinguishing it from previously described species within this genus. Based on the results herein reported, the new South-American species Paraboeremia yungensis is described and proposed.

Multigene Phylogeny and Morphology Reveal Unexpectedly High Number of New Species of Cantharellus Subgenus Parvocantharellus (Hydnaceae, Cantharellales) in China.

The genus Cantharellus, commonly known as chanterelles, has recently been divided into six subgenera; however, wider sampling approaches are needed to clarify the relationships within and between these groups. A phylogenetic overview of Cantharellus subgenus Parvocantharellus in China was inferred based on the large subunit nuclear ribosomal RNA gene (nrLSU), the DNA-directed RNA polymerase II subunit 2 (rpb2), and the transcription elongation factor 1-alpha (tef1). A total of nine species from China were assigned to the subgenus, including seven novel species, namely Cantharellus aurantinus, C. austrosinensis, C. galbanus, C. luteolus, C. luteovirens, C. minioalbus, and C. sinominior, and two known species, namely C. albus and C. zangii. The detailed descriptions and illustrations were provided based on the newly obtained data, with the comparisons to closely related species. C. zangii was restudied based on the paratype specimens and multiple new collections from the type locality. Futhermore, the Indian species C. sikkimensis was identified as a synonym of C. zangii based on the morphological and molecular analyses. A key to the Chinese species belonging to the subg. Parvocantharellus is also provided.

Phylogeography and Taxonomic Revision of the Pen Shell Atrina pectinata Species Complex in the South China Sea

Phylogeographic studies contribute to addressing questions regarding the geographic patterns and evolutionary scenarios within and among species and also shed light on the taxonomic status of widely distributed species complexes. The pen shell Atrina pectinata species complex is a widely distributed and economically important bivalve in the northwestern Pacific. Previous phylogeographic studies have identified four genetically distinct cryptic species within the A. pectinata species complex along the coast of China, of which three cryptic species were distributed in the South China Sea. However, less attention has been given to their identification and delimitation. Herein, we report the phylogeography and taxonomic revision of the A. pectinata species complex in the South China Sea using DNA sequence data from mitochondrial cytochrome oxidase I (COI) and 16S ribosomal RNA (16S rRNA), and nuclear 28S ribosomal RNA (28S rRNA) gene markers. Using a combination of phylogenetic and DNA-based species delimitation analysis methods, we found strong support for four genetically valid species in the A. pectinata species complex and defined them as A. japonica, A. lischkeana, Atrina sp., and A. pectinata based on our results as well as on previous morphological and genetic studies. A demographic historical analysis showed that all three species in the South China Sea had populations that were relatively stable over time and then subjected to sudden expansion during the late Pleistocene (60,000–90,000 years ago). These results provide new insights into the systematics and evolution of the A. pectinata species complex and have important conservation and management implications.

Taxonomy and Phylogeny of the Favolaschia calocera Complex (Mycenaceae) with Descriptions of Four New Species

Favolaschia calocera was originally described from Madagascar, and reported to have a worldwide distribution. In the current study, samples of the Favolaschia calocera from Central America, Australia, China, Kenya, Italy, New Zealand, and Thailand were analyzed by using both morphological and molecular methods. Phylogenetic analyses were based on the internal transcribed spacer (ITS) dataset, and the combined five-locus dataset of ITS, large subunit nuclear ribosomal RNA gene (nLSU), the small subunit mitochondrial rRNA gene (mt-SSU), the small subunit of nuclear ribosomal RNA gene (nu-SSU), and the translation elongation factor 1α (TEF1). Our study proves that Favolaschia calocera is a species complex, and six species are recognized in the complex including four new species. Three new species F. brevibasidiata, F. brevistipitata, and F. longistipitata from China; and one new species F. minutissima from Asia. In addition, Favolaschia claudopus (Singer) Q.Y. Zhang & C. Dai, earlier treated as a variety of Favolaschia calocera R. Heim, were raised to species rank. Illustrated descriptions of these five new taxa are given. An identification key and a comparison of the characteristics of species in the Favolaschia calocera complex are provided.

Multi-Gene Phylogeny and Taxonomy of Hydnellum (Bankeraceae, Basidiomycota) from China.

The genus Hydnellum is an important group of stipitate hydnaceous fungi which can form ectomycorrhiza with many species of woody plants. In recent decades, the frequency and number of basidiocarps observed in China have been declining significantly. So far, however, we know little about the species diversity of Hydnellum in China. In this study, we conducted molecular phylogenetic analyses based on sections of multiple loci, including the large subunit of nuclear ribosomal RNA gene (nLSU), the internal transcribed spacer regions (ITS), the small subunit of nuclear ribosomal RNA gene (SSU) and the second-largest subunit of RNA polymerase II gene (RPB2), as well as morphological studies, of collected samples of Hydnellum from China. We also inferred Maximum Likelihood and Bayesian phylogenies for the order Thelephorales from the dataset of the combined nLSU and ITS. This study has revealed the phylogenetic position of Hydnellum in the order Thelephorales, and phylogenetically confirmed ten major clades in Thelephorales; Twenty-nine taxa are proposed, described or reported, including 10 new subgenera (Hydnellum subgenus Hydnellum, subg. Caesispinosum, subg. Croceum, subg. Inflatum, subg. Rhizomorphum, subg. Scabrosum, subg. Spongiosum, subg. Subindufibulatum, subg. Violaceum and subg. Zonatum), 11 new species (Hydnellum atrorubrum, H. atrospinosum, H. bomiense, H. brunneorubrum, H. fibulatum, H. granulosum, H. inflatum, H. rubidofuscum, H. squamulosum, H. sulcatum and H. yunnanense), 3 newly recorded species (H. caeruleum, H. peckii and H. spongiosipes) and 5 notable specimens (Hydnellum sp 1, H. sp 2, H. sp 3, H. sp 4 and H. sp 5). A classification system based on the morphological characteristics (especially the hyphal structure types) and molecular analyses is proposed to accommodate most species in Hydnellum. The distinguishing characters of the subgenera and the new species with their closely related taxa are discussed. A key to the species of Hydnellum from China is provided.

Disporella guada sp. nov., an erect-ramose rectangulate cyclostome (Bryozoa, Stenolaemata) from the Caribbean Sea: convergent evolution in bryozoan colony morphology

The taxonomy of cyclostome bryozoans is founded on characters of the skeleton, but molecular sequence data have increasingly shown that established higher taxa are not monophyletic. Here we describe the skeletal morphology of a new species from Guadeloupe (French West Indies) with erect ramose colonies consisting of long, curved zooids that are typical of the suborder Cerioporina among living cyclostomes. However, molecular evidence from nuclear ribosomal RNA genes 18S and 28S places the new taxon in the suborder Rectangulata, where this colony-form has not been previously recorded. It nests firmly within the genus Disporella Gray, 1848, in a strongly supported clade that also includes Plagioecia patina (Lamarck, 1816) (Tubuliporina) and the sister taxa Doliocoitis cyanea Gordon & Taylor, 2001 (Rectangulata) and Favosipora rosea Gordon & Taylor, 2001 (Cerioporina). The short and robust branches of the new Guadeloupe cyclostome, here named Disporella guada Harmelin, Taylor & Waeschenbach sp. nov., are well adapted to life in shallow rocky sites exposed to severe wave action, which appear to be its exclusive habitat.

Redescription of Biacetabulum giganteum Hunter, 1929 (Cestoda: Caryophyllidea), description of two new, closely related species from suckers (Catostomidae) in North America, and a critical review of host specificity of species of Biacetabulum Hunter, 1927

Morphological and molecular evaluation of tapeworms of the order Caryophyllidea from suckers (Cypriniformes: Catostomidae) in North America made it possible to redescribe the insufficiently known and rare species Biacetabulum giganteum Hunter, 1929, based on specimens from the type host, small mouth buffalo, Ictiobus bubalus. This cestode is mainly typified by an elongate bottle-shaped body with a narrow, long neck region distinctly separating a relatively large scolex with deep loculi and a sharply widening body. Molecular phylogenetic analysis based on sequences of the large subunits of the nuclear ribosomal RNA genes (lsrDNA or 28S rDNA) placed B. giganteum in a clade called herein the B. giganteum-species complex, which also includes two morphologically similar new species. Biacetabulum magdae sp. nov. is described from the blacktail redhorse, Moxostoma poecilurum (type host), and the spotted sucker, Minytrema melanops, in Mississippi, USA. It differs from B. giganteum by the scolex with a weakly developed apical disc and shallow median and lateral loculi. Biacetabulum johni sp. nov. is described from Minytrema melanops (type host) in Florida and Moxostoma poecilurum in Mississippi, USA. Biacetabulum johni sp. nov. can be distinguished from B. magdae sp. nov. by a smaller size of the body (5–7 mm versus 10–12 mm) and a relatively longer neck, with the first vitelline follicles beginning at about the mid-length of the body in B. johni (versus at the first third or fourth of the body in B. magdae). With two new species described herein, Biacetabulum becomes the most species-rich genus of caryophyllidean tapeworms, with a total of 14 nominal species known to date (recently described B. hypentelii is synonymized with B. longicollum based on newly generated molecular data). Host specificity of species of Biacetabulum Hunter, 1927 was critically reviewed and it is shown that the actual host specificity is considerably higher than that reported in the literature. http://zoobank.org/urn:lsid:zoobank.org:pub:lsid:zoobank.org:pub:ABF5446A-F170-4CA0-9AF7-D5234BD71139

Two new brown rot polypores from tropical China.

Brown-rot fungi are types of fungi that selectively degrade cellulose and hemicellulose from wood and are perhaps the most important agents involved in the degradation of wood products and dead wood in forest ecosystem. Two new brown-rot species, collected from southern China, are nested within the clades of Fomitopsis sensu stricto and Oligoporus sensu stricto, respectively. Their positions are strongly supported in the Maximum Likelihood phylogenetic tree of the concatenated the internal transcribed spacer (ITS) regions, the large subunit of nuclear ribosomal RNA gene (nLSU), the small subunit of nuclear ribosomal RNA gene (nuSSU), the small subunit of mitochondrial rRNA gene (mtSSU), the largest subunit of RNA polymerase II (RPB1), the second largest subunit of RNA polymerase II (RPB2) and the translation elongation factor 1-α gene (TEF1) sequences. Fomitopsisbambusae, only found on bamboo, is characterised by its resupinate to effused-reflexed or pileate basidiocarps, small pores (6–9 per mm), the absence of cystidia, short cylindrical to oblong-ellipsoid basidiospores measuring 4.2–6.1 × 2–2.3 μm. Oligoporuspodocarpi is characterised by white to pale cream pore surface, round or sometimes angular pores (5–6 per mm), broadly ellipsoid to reniform basidiospores measuring 3.8–4.2 × 2–2.3 μm and growing on Podocarpus. Illustrated descriptions of these two novel species, Fomitopsisbambusae and Oligoporuspodocarpi, are provided.

Phlebiopsis xuefengensis sp. nov. from Gastrodia elata (Orchidaceae) in Hunan Province, Southern China

Phlebiopsis is a genus of poroid crust fungi belong to the family Phanerochaetaceae. The genus contains 19 species, which show a widespread distribution. In this paper, a strain of Phlebiopsis sp., named WS01, was isolated from tubers of Gastrodia elata collected in southern China. The endophytic fungus WS01 bearing fruiting-body was cultivated successfully in the fruiting body media. The characterized strain, in hyphae of the hymenium with colorless walls and cystidia lacking revealed that it morphologically differs from other Phlebiopsis species. A combined dataset of the internal transcribed spacer (ITS), the large subunit regions of nuclear ribosomal RNA gene (LSU) and RNA polymerase II largest subunit gene (rpb1) was used to confirm the phylogenetic placement of this species, and phylogenetic analyses were carried out with maximum likelihood and bayesian inference methods. The results provide evidence that the species represents a new member of the Phlebiopsis clade in the family Phanerochaetaceae, in which the new species formed a monophyletic lineage with a strong support (100% ML, 1.00 BPP) and then grouped with P. pilatii. According to a signature sequence for a new species, the strain is, therefore, proposed as Phlebiopsis xuefengensis sp. nov. It is the first record of Phlebiopsis fungi from the plant endophytes and the first novel fungal species isolated from Gastrodia elata.

A new species of Albatrellus sensu stricto (Albatrellaceae, Russuales) from China

Based on morphological and molecular data, a new species of Albatrellus sensu stricto, A. alpinus sp. nov., is described from Yunnan Province of southwestern China. Albatrellus alpinus is characterized by a buff to pale yellow pileal surface, simple septate hyphae, yellow and shapeless oily substance present in all hyphae but seldom in tramal hyphae, and small amyloid basidiospores. Phylogenetic analyses based on the nuclear ribosomal internal transcribed spacer (nrITS) and the large subunit of the nuclear ribosomal RNA gene (nLSU) indicated that the new species is nested within Albatrellus sensu stricto. A key to the Chinese species of Albatrellus sensu stricto is provided.

A phylogenetic overview of the Hydnaceae (Cantharellales, Basidiomycota) with new taxa from China.

The family Hydnaceae (Cantharellales, Basidiomycota) is a group of fungi found worldwide which exhibit stichic nuclear division. The group is highly diverse in morphology, ecology, and phylogeny, and includes some edible species which are popular all over the world. Traditionally, Hydnaceae together with Cantharellaceae, Clavulinaceae and Sistotremataceae are four families in the Cantharellales. The four families were combined and redefined as “Hydnaceae”, however, a comprehensive phylogeny based on multiple-marker dataset for the entire Hydnaceae sensu stricto is still lacking and the delimitation is also unclear. We inferred Maximum Likelihood and Bayesian phylogenies for the family Hydnaceae from the data of five DNA regions: the large subunit of nuclear ribosomal RNA gene (nLSU), the internal transcribed spacer regions (ITS), the mitochondrial small subunit rDNA gene (mtSSU), the second largest subunit of RNA polymerase II (RPB2) and the translation elongation factor 1-alpha gene (TEF1). We also produced three more phylogenetic trees for Cantharellus based on 5.8S, nLSU, mtSSU, RPB2 and TEF1, Craterellus and Hydnum both based on the combined nLSU and ITS. This study has reproduced the status of Hydnaceae in the order Cantharellales, and phylogenetically confirmed seventeen genera in Hydnaceae. Twenty nine new taxa or synonyms are described, revealed, proposed, or reported, including eight new subgenera (Cantharellus subgenus Magnus, Craterellus subgenus Cariosi, subg. Craterellus, subg. Imperforati, subg. Lamelles, subg. Longibasidiosi, subg. Ovoidei, and Hydnum subgenus Brevispina); seventeen new species (Ca. laevihymeninus, Ca. magnus, Ca. subminor, Cr. badiogriseus, Cr. croceialbus, Cr. macrosporus, Cr. squamatus, H. brevispinum, H. flabellatum, H. flavidocanum, H. longibasidium, H. pallidocroceum, H. pallidomarginatum, H. sphaericum, H. tangerinum, H. tenuistipitum and H. ventricosum); two synonyms (Ca. anzutake and Ca. tuberculosporus as Ca. yunnanensis), and two newly recorded species (H. albomagnum and H. minum). The distinguishing characters of the new species and subgenera as well as their allied taxa are discussed in the notes which follow them. The delimitation and diversity in morphology, ecology, and phylogeny of Hydnaceae is discussed. Notes of seventeen genera which are phylogenetically accepted in Hydnaceae by this study and a key to the genera in Hydnaceae are provided.

High-throughput sequencing outperforms traditional morphological methods in Blue Catfish diet analysis and reveals novel insights into diet ecology.

Blue Catfish Ictalurus furcatus are an invasive, yet economically important species in the Chesapeake Bay. However, their impact on the trophic ecology of this system is not well understood. In order to provide in‐depth analysis of predation by Blue Catfish, we identified prey items using high‐throughput DNA sequencing (HTS) of entire gastrointestinal tracts from 134 samples using two genetic markers, mitochondrial cytochrome c oxidase I (COI) and the nuclear 18S ribosomal RNA gene. We compared our HTS results to a more traditional “hybrid” approach that coupled morphological identification with DNA barcoding. The hybrid study was conducted on additional Blue Catfish samples (n = 617 stomachs) collected from the same location and season in the previous year. Taxonomic representation with HTS vastly surpassed that achieved with the hybrid methodology in Blue Catfish. Significantly, our HTS study identified several instances of at‐risk and invasive species consumption not identified using the hybrid method, supporting the hypothesis that previous studies using morphological methods may greatly underestimate consumption of critical species. Finally, we report the novel finding that Blue Catfish diet diversity inversely correlates to daily flow rates, perhaps due to higher mobility and prey‐seeking behaviors exhibited during lower flow.

Unique environmental Symbiodiniaceae diversity at an isolated island in the northwestern Pacific

Dinoflagellates in the family Symbiodiniaceae are intensively investigated as algal symbionts of corals and other invertebrates, underpinning coral reef ecosystems as primary producers. Diversity, including regional diversification, of free-living communities is less studied. In this study, an environmental Symbiodiniaceae community at an isolated island, Okinotori Island, Japan, was investigated to determine whether the community is endemic or common with other locations near continents and major ocean currents. Symbiotic algae in common corals at the island were the same type as those of the corals from other Japanese waters. In the environmental samples, genera Symbiodinium (formerly clade A), Cladocopium (clade C), Durusdinium (clade D), and clades F (including Freudenthalidium), G, and I, were identified through analysis of internal transcribed spacer region 2 of nuclear ribosomal RNA gene (ITS2) sequences. Interestingly, some sequences found were genetically different from those of previously reported genera/clades. These unknown sequences were genetically included in the Symbiodiniaceae linage, but they were differentiated from the previously known nine clades. The sequences formed a cluster in the phylogenetic tree based on 28S nrDNA. These sequences were thus considered members of a novel clade in the family (clade J). In total, 120 kinds of ITS2 sequences were produced; while 10 were identical to previously reported sequences, the majority were highly divergent. These genetically unique Symbiodiniaceae types, including novel clade J, may have evolved in isolation and reflect the environmental characteristics of the Okinotori Island.

Phylogenetic analysis of the distribution of deadly amatoxins among the little brown mushrooms of the genus Galerina.

Some but not all of the species of 'little brown mushrooms' in the genus Galerina contain deadly amatoxins at concentrations equaling those in the death cap, Amanita phalloides. However, Galerina's ~300 species are notoriously difficult to identify by morphology, and the identity of toxin-containing specimens has not been verified with DNA barcode sequencing. This left open the question of which Galerina species contain toxins and which do not. We selected specimens for toxin analysis using a preliminary phylogeny of the fungal DNA barcode region, the ribosomal internal transcribed spacer (ITS) region. Using liquid chromatography/mass spectrometry, we analyzed amatoxins from 70 samples of Galerina and close relatives, collected in western British Columbia, Canada. To put the presence of toxins into a phylogenetic context, we included the 70 samples in maximum likelihood analyses of 438 taxa, using ITS, RNA polymerase II second largest subunit gene (RPB2), and nuclear large subunit ribosomal RNA (LSU) gene sequences. We sequenced barcode DNA from types where possible to aid with applications of names. We detected amatoxins only in the 24 samples of the G. marginata s.l. complex in the Naucoriopsis clade. We delimited 56 putative Galerina species using Automatic Barcode Gap Detection software. Phylogenetic analysis showed moderate to strong support for Galerina infrageneric clades Naucoriopsis, Galerina, Tubariopsis, and Sideroides. Mycenopsis appeared paraphyletic and included Gymnopilus. Amatoxins were not detected in 46 samples from Galerina clades outside of Naucoriopsis or from outgroups. Our data show significant quantities of toxin in all mushrooms tested from the G. marginata s.l. complex. DNA barcoding revealed consistent accuracy in morphology-based identification of specimens to G. marginata s.l. complex. Prompt and careful morphological identification of ingested G. marginata s.l. has the potential to improve patient outcomes by leading to fast and appropriate treatment.