Small-subunit Sequences Research Articles

First report of Colletotrichum viniferum causing leaf spot of Hopea odorata in Bangladesh

An outbreak of leaf spot disease affecting Hopea odorata was detected during a survey conducted in September 2019 in two different plantations in Sylhet, Bangladesh. The spots on leaves were small, dark brown to blackish and circular to irregular in shape (Fig. 1). They enlarged progressively in size with the maturity of leaves. Diseased leaves were collected, cut into small pieces (5 × 5 mm), treated with sodium hypochlorite (1%) solution for 30 seconds, and then rinsed with sterile distilled water three times. After drying on filter paper, the pieces were placed aseptically on potato dextrose agar (PDA) amended with kanamycin monosulphate and ampicillin, each at 50 μg/ml. The plates were incubated at 25°C for four days in the dark to allow fungal growth. The fungal mycelia were sub-cultured several times on the same media amended with the same amount of antibiotics to obtain pure cultures. Six-day-old colonies on PDA had greyish-white, dense, cottony mycelia (Fig. 2). Mycelial growth was optimal at 25°C (radial growth rate, 8.2 mm/d), whereas no growth was observed at 32°C. Conidia were single- celled, smooth-walled and cylindrical with rounded ends, the length and width ranged from 14 to 18 μm (mean, 15.6 ±1.2 μm, n=50) and 4.5 to 5.5 μm (mean, 4.8 ±0.5 μm, n=50) respectively (Fig. 3). Conidiophores were unbranched and hyaline. Setae were absent but appressoria were observed with various shapes (e.g., ovoid, globose and clavate). All these morphological features suggested that the species was likely to Collectotrichum viniferum as described by Peng et al. (4). For molecular identification, genomic DNA of one representative isolate was extracted using a genomic DNA extraction kit (Favorgen Biotech Corporation, Taiwan). The complete sequences of the internal transcribed spacer 1 (ITS1) and 5.8S ribosomal DNA (rDNA), and the partial sequences of ITS2 and small subunit of the fungal rDNA were amplified using primers SR6R (forward) and LR1 (reverse) as described by Meyer et al. (3). A BLAST search with the amplified sequence and a phylogenetic tree revealed that the sequence (GenBank Accession No. MT876535.2) was identical to C. viniferum (KX594334) (Fig. 4). The phylogenetic tree was constructed using MEGA7 software (Kumar et al., 2) as described previously (Azad et al., 1). A pathogenicity test was performed using ten detached healthy leaves which were inoculated with 0.5 ml of a 105 conidia/ml spore suspension. Sterile distilled water was used as a control. Inoculated leaves were incubated in a growth chamber at 25°C (±2°C) with a 14-hr photoperiod. Seven days after inoculation, black necrotic spots appeared on the inoculated leaves, whereas the control leaves showed no disease symptoms (Fig. 5). C. viniferum was re-isolated from the necrotic spots of inoculated leaves and re-identified, thus fulfilling Koch's postulates. As per our knowledge, this is the first report on C. viniferum causing leaf spot of H. odorata in Bangladesh. Further studies and surveys are essential to determine the severity of the disease in different regions of the country, and provide a detailed characterisation of the pathogen. The authors are thankful to the Research Centre, Shahjalal University of Science and Technology, Bangladesh, for allocating fund to conduct the study (grant no. FES/2018/2/05).

Parvilucifera multicavata sp. nov. (Alveolata, Perkinsozoa), a New Parasitoid Infecting Marine Dinoflagellates Having Abundant Apertures on the Sporangium

The phylum Perkinsozoa is known as an exclusively parasitic group including the parasites of shellfish, fish, dinoflagellates, cryptophytes, and tadpoles and at present comprises seven genera across three families (Parviluciferaceae, Perkinsidae, and Xcellidae), with the genus Parvilucifera having the most abundant species in the family Parviluciferaceae. During intensive sampling along the Korean coast in August and September 2017, a new species of the genus Parvilucifera was discovered and successfully established in cultures. Morphological and ultrastructural observations revealed that the new parasitoid shares almost all known diagnostic characters with other species of Parvilucifera, except that its sporangium has a higher number of apertures although with smaller diameters than those in P. infectans. Molecular phylogenetic trees based on both nuclear small subunit (SSU) and concatenated SSU and large subunit (LSU) ribosomal DNA (rDNA) sequences revealed that the new parasitoid was nested within the family Parviluciferaceae and had a sister relationship with P. infectans. Based on morphological, ultrastructural, and molecular data, we propose to erect a new species, P. multicavata sp. nov., for the new parasitoid found in this study.

A new lichen-forming fungus, Orientophila corticola, from South Korea, with a key to the genus

Abstract Orientophila corticola is described as a new lichen-forming fungus from South Korea. The new species is distinguishable from other Orientophila species by the substrate preference to tree barks and the habitat specificity to inland areas. Molecular analyses applying internal transcribed spacer (ITS) and mitochondrial small subunit (mtSSU) sequences clearly classify O. corticola as a distinct species in the genus Orientophila. A surrogate key is provided to assist in the identification of all 13 taxa in the genus Orientophila.

Marine mammal skin microbiotas are influenced by host phylogeny.

Skin-associated microorganisms have been shown to play a role in immune function and disease of humans, but are understudied in marine mammals, a diverse animal group that serve as sentinels of ocean health. We examined the microbiota associated with 75 epidermal samples opportunistically collected from nine species within four marine mammal families, including: Balaenopteridae (sei and fin whales), Phocidae (harbour seal), Physeteridae (sperm whales) and Delphinidae (bottlenose dolphins, pantropical spotted dolphins, rough-toothed dolphins, short-finned pilot whales and melon-headed whales). The skin was sampled from free-ranging animals in Hawai‘i (Pacific Ocean) and off the east coast of the United States (Atlantic Ocean), and the composition of the bacterial community was examined using the sequencing of partial small subunit (SSU) ribosomal RNA genes. Skin microbiotas were significantly different among host species and taxonomic families, and microbial community distance was positively correlated with mitochondrial-based host genetic divergence. The oceanic location could play a role in skin microbiota variation, but skin from species sampled in both locations is necessary to determine this influence. These data suggest that a phylosymbiotic relationship may exist between microbiota and their marine mammal hosts, potentially providing specific health and immune-related functions that contribute to the success of these animals in diverse ocean ecosystems.

Phylogeny and cultivation of the holocarpic oomycete Diatomophthora perforans comb. nov., an endoparasitoid of marine diatoms

Oomycetes infecting diatoms are biotrophic parasitoids and live in both marine and freshwater environments. They are ubiquitous, but the taxonomic affinity of many species remains unclear and the majority of them have not been studied for their molecular phylogeny. Only recently, the phylogenetic and taxonomic placement of some diatom-infecting, early-diverging oomycetes was resolved, including the genera Ectrogella, Miracula, Olpidiopsis, and Pontisma. A group of holocarpic diatom parasitoids with zoospores swarming within the sporangium before release were found to be unrelated to the known genera with diatom-infecting species, and were re-classified to a new genus, Diatomophthora. However, about a dozen species of holocarpic diatom parasitoids with unclear affinity remained unsequenced, which includes a commonly occurring species so far identified as Ectrogella perforans. However, this assignment to Ectrogella is doubtful, as the species was not reported to feature a clear-cut diplanetism, a hallmark of Ectrogella s. str. and the whole class Saprolegniomycetes. It was the aim of the current study to clarify the phylogenetic affinities of the species and if the rather broad host range reported is correct or a reflection of cryptic species. By targeted screening, the parasitoid was rediscovered from Helgoland Roads, North Sea and Oslo Fjord, Southern Norway and investigated for its phylogenetic placement using small ribosomal subunit (18S) sequences. Stages of its life cycle on different marine diatoms were described and its phylogenetic placement in the genus Diatomophthora revealed. A stable host-parasite axenic culture from single spore strains of the parasitoid were established on several strains of Pleurosigma intermedium and Coscinodiscus concinnus. These have been continuously cultivated along with their hosts for more than 2 years, and cultural characteristics are reported. Cross-infection trials revealed the transferability of the strains between hosts under laboratory conditions, despite some genetic distance between the pathogen strains. Thus, we hypothesise that D. perforans might be in the process of active radiation to new host species.

Molecular and morphological characterization of Ektaphelenchoides maafiae n. sp. (Nematoda: Ektaphelenchinae) from northern forests of Iran

AbstractEktaphelenchoides maafiae n. sp. was isolated during a survey of nematodes associated with bark samples of an oak tree (Quercus castaneifolia) in Gorgan, Golestan Province, northern Iran. The new species has a body length of 480–546 μm (in females) and 431–480 μm (in males). The cuticle is weakly annulated, with three lateral lines. Lip region offset. The stylet is 13–15 μm long without basal swellings. The excretory pore is located at the level of the metacorpus base to slightly posterior, and hemizonid is at 15–17 μm posterior to the excretory pore. The post‐uterine sac is short, 6–8 μm long. Spicules having rounded condylus, rostrum short, conical with bluntly pointed tip, a cucullus (apophysis) presented on the dorsal distal end. Male tail bearing four (2 + 2) caudal papillae, conical, with sharply pointed terminus. The new species is close to four known species of the genus, including E. hunti, E. ruehmi, E. caspiensis and E. poinari, but differs from them by body size, shape of tail terminus, stylet length, shape and size of spicules, length of post‐vulval uterine sac and number of caudal papillae. Phylogenetic analysis based on small subunit (SSU) and partial large subunit (LSU) sequences of rDNA confirmed its status as a new species.

Rhizoglomus dalpeae, R. maiae, and R. silesianum, new species

ABSTRACTWe examined three arbuscular mycorrhizal fungi (AMF; phylum Glomeromycota) producing glomoid spores. The mode of formation and morphology of these spores suggested that they represent undescribed species in the genus Rhizoglomus of the family Glomeraceae. Subsequent morphological studies of the spores and molecular phylogenetic analyses of sequences of the nuc rDNA small subunit (18S), internal transcribed spacer (ITS1-5.8S-ITS2 = ITS), and large subunit (28S) region (= 18S-ITS-28S) confirmed the suggestion and indicated that the fungi strongly differ from all previously described Rhizoglomus species with known DNA barcodes. Consequently, the fungi were described here as new species: R. dalpeae, R. maiae, and R. silesianum. Two of these species lived hypogeously in the field in habitats subjected to strong environmental stresses. Rhizoglomus dalpeae originated from an inselberg located within Guineo-Sudanian transition savanna zone in Benin, West Africa, where the temperature of the inselberg rock during a 5-mo drought ranges from 40 to 60 C. Rhizoglomus silesianum originated from a coal mine spoil heap in Poland, whose substrate is extremely poor in nutrients, has unfavorable texture, and may heat up to 50 C. By contrast, R. maiae was found in more favorable habitat conditions. It produced an epigeous cluster of spores among shrubs growing in a tropical humid reserve in Brazil. Moreover, the compatibility of phylogenies of species of the family Glomeraceae reconstructed from analyses of sequences of 18S-ITS-28S and the largest subunit of RNA polymerase II (RPB1) gene was discussed.

Experimental warming alters the community composition, diversity, and N2 fixation activity of peat moss (Sphagnum fallax) microbiomes.

Sphagnum‐dominated peatlands comprise a globally important pool of soil carbon (C) and are vulnerable to climate change. While peat mosses of the genus Sphagnum are known to harbor diverse microbial communities that mediate C and nitrogen (N) cycling in peatlands, the effects of climate change on Sphagnum microbiome composition and functioning are largely unknown. We investigated the impacts of experimental whole‐ecosystem warming on the Sphagnum moss microbiome, focusing on N2 fixing microorganisms (diazotrophs). To characterize the microbiome response to warming, we performed next‐generation sequencing of small subunit (SSU) rRNA and nitrogenase (nifH) gene amplicons and quantified rates of N2 fixation activity in Sphagnum fallax individuals sampled from experimental enclosures over 2 years in a northern Minnesota, USA bog. The taxonomic diversity of overall microbial communities and diazotroph communities, as well as N2 fixation rates, decreased with warming (p < 0.05). Following warming, diazotrophs shifted from a mixed community of Nostocales (Cyanobacteria) and Rhizobiales (Alphaproteobacteria) to predominance of Nostocales. Microbiome community composition differed between years, with some diazotroph populations persisting while others declined in relative abundance in warmed plots in the second year. Our results demonstrate that warming substantially alters the community composition, diversity, and N2 fixation activity of peat moss microbiomes, which may ultimately impact host fitness, ecosystem productivity, and C storage potential in peatlands.

Rediscovery and phylogenetic placement of Olpidiopsis gillii (de Wildeman) Friedmann, a holocarpic oomycete parasitoid of freshwater diatoms

Abstract The genus Olpidiopsis of the Oomycota includes several species that are aquatic parasites and hyperparasites. Despite their widespread occurrence and potential ecological importance, only a handful of these species has been subjected to phylogenetic investigations, so far. Most species have not been observed and reported for several decades. In the current study, the freshwater diatom parasite Olpidiopsis gillii (de Wild.) Friedmann was rediscovered from the river Main in Germany and investigated for its phylogenetic placement using nuclear small ribosomal subunit (SSU) sequences. The absence of a zoospore diplanetism is a characteristic of the genus Olpidiopsis, which is in contrast to the diplanetism observed in species of Ectrogella. The phylogenetic reconstruction revealed that Olpidiopsis gillii is a basal lineage within the oomycetes, grouping together with the recently-described marine diatom parasite Olpidiopsis drebesii with high support, and loosely associated with Olpidiopsis species parasitising red algae. However, as there are no sequence data available for the type species of both Olpidiopsis and Ectrogella the taxonomic assignment of these simple holocarpic parasites of algae and diatoms remains fraught with uncertainty.

Description of Ektaphelenchus cupressi n. sp. (Nematoda: Ektaphelenchinae) from Iran

Summary Ektaphelenchus cupressi n. sp. was isolated during a survey of nematodes associated with bark samples of a dead cypress tree (Cupressus sempervirens) in Mazandaran province, northern Iran. The new species is characterised by a female body length of 612 (520-693) μm, stylet 17-19 μm long with wide lumen and lacking basal swellings, head region hemispherical in lateral view and slightly offset from the body contour by a shallow constriction, three incisures in the lateral field, excretory pore situated anterior to level of metacorpus valve, hemizonid not seen, post-uterine sac 29-35 μm long, shape of the tail terminus, and arrangement of the male caudal papillae. By a combination of morphological characters, e.g., stylet without knobs or swellings at the base, short conical tail, PUS length, and only a vestigial anus and rectum in most individuals, the new species shares similarities with species belonging to three genera, i.e., Anomyctus, Ektaphelenchus, and Seinura. Phylogenetic analysis based on small subunit (SSU) and partial large subunit (LSU) sequences of rDNA confirmed its status as a new species.

Molecular phylogenetic study of Bealius Massey &amp; Hinds, 1970 (Nematoda: Hexatylina: Sphaerulariidae), with description of B. pinus n. sp. from Iran

Summary Bealius pinus n. sp. is described and illustrated based on its morphological, morphometric, and molecular characters. The new species is characterised by a body length of 1040 (920-1124) μm (females) and 985 (990-1043) μm (males), lip region continuous, 7.2 (7.0-8.0) μm broad, 3.3 (3.0-4.0) μm high, stylet length 8.4 (7.0-9.0) μm or ca 1.0-1.3 times the lip region diam., lateral fields with five or six incisures, the outer incisures crenate and inner incisures weakly crenate, and excretory pore situated 156 (141-170) μm from the anterior end. The basal pharyngeal bulb has a short posterior extension projecting dorsally into the intestine. Post-uterine sac absent. Functional males common in the population, spicules 29 (28-30) μm long. Tail cylindrical with broadly rounded terminus in the female and conical, arcuate, completely enveloped by a well-developed bursa in the male. The new species is compared with two known species of the genus, B. bisulcus and B. pissodi. Molecular phylogenetic studies of the new species using partial sequences of small subunit (SSU) rDNA revealed that it forms a clade with several nematode species belonging to genera in the Sphaerularioidea, namely: Sphaerularia, Paurodontella, Nothotylenchus, and Deladenus.

Vegetative reproduction and phylogeny of a new subtidal alga from Korea: Dipterocladia yongdeokkoi sp. nov. (Ceramiales, Rhodophyta)

ABSTRACTDipterocladia yongdeokkoi sp. nov. was collected at depths of 6–15 m at three locations at Jeju Island, Korea: Biyangdo, Hado and Shinchang. All plants were vegetative without reproductive structures. For two years in laboratory culture, plants were consistently vegetative. Cultured plants formed single-celled diaspores and multicellular propagules that grew into mature plants. Dipterocladia yongdeokkoi is distinguished by its flattened thallus arising from a thick, cylindrical stipe, compressed axes, irregular branches and alternately pinnate determinate adventitious laterals and pseudolaterals. Analyses of mitochondrial cox1, plastid rbcL, and nuclear small subunit (SSU) sequences support the monophyly of Dipterocladia and the distinctness of D. yongdeokkoi from other species of Dipterocladia.

New operational taxonomic units of Enterocytozoon in three marsupial species

BackgroundEnterocytozoon bieneusi is a microsporidian, commonly found in animals, including humans, in various countries. However, there is scant information about this microorganism in Australasia. In the present study, we conducted the first molecular epidemiological investigation of E. bieneusi in three species of marsupials (Macropus giganteus, Vombatus ursinus and Wallabia bicolor) living in the catchment regions which supply the city of Melbourne with drinking water.MethodsGenomic DNAs were extracted from 1365 individual faecal deposits from these marsupials, including common wombat (n = 315), eastern grey kangaroo (n = 647) and swamp wallaby (n = 403) from 11 catchment areas, and then individually tested using a nested PCR-based sequencing approach employing the internal transcribed spacer (ITS) and small subunit (SSU) of nuclear ribosomal DNA as genetic markers.ResultsEnterocytozoon bieneusi was detected in 19 of the 1365 faecal samples (1.39%) from wombat (n = 1), kangaroos (n = 13) and wallabies (n = 5). The analysis of ITS sequence data revealed a known (designated NCF2) and four new (MWC_m1 to MWC_m4) genotypes of E. bieneusi. Phylogenetic analysis of ITS sequence data sets showed that MWC_m1 (from wombat) clustered with NCF2, whereas genotypes MWC_m2 (kangaroo and wallaby), MWC_m3 (wallaby) and MWC_m4 (kangaroo) formed a new, divergent clade. Phylogenetic analysis of SSU sequence data revealed that genotypes MWC_m3 and MWC_m4 formed a clade that was distinct from E. bieneusi. The genetic distinctiveness of these two genotypes suggests that they represent a new species of Enterocytozoon.ConclusionsFurther investigations of Enterocytozoon spp. from macropods and other animals will assist in clarifying the taxonomy and epidemiology of these species in Australia and elsewhere, and in assessing the public health risk of enterocytozoonosis.

Morphology, morphogenesis, and phylogeny of an Anteholosticha intermedia (Ciliophora, Urostylida) population from the United States

A distinct population of Anteholosticha intermedia was isolated from soil in the Great Smoky Mountains of North Carolina, USA, and its morphology, morphogenesis and molecular phylogeny investigated by microscopic observations of live and protargol-prepared specimens and analyses of the sequence of small subunit (SSU) rDNA. Our population closely resembles the populations from Austria and Korea. Members of the genus Anteholosticha have been regarded as ontogenetically diverse, which is confirmed by the present work. The most noteworthy ontogenetic feature of the American population of A. intermedia is that the oral primordium in the proter appears apokinetally at the posterior end of the undulating membranes anlage at the beginning of division and then dedifferentiates midway through morphogenesis. Molecular phylogenetic analyses demonstrate, with high support, that the American population of A. intermedia is clearly distinct from congeners and branches as part of a sister lineage to the Bakuella–Urostyla clade that belongs to the major clade comprising the order Urostylida.

Biological and Molecular Characterization of Trypanosoma cruzi Strains from Four States of Brazil.

Chagas disease affects between six and seven million people. Its etiological agent, Trypanosoma cruzi, is classified into six discrete typing units (DTUs). The biological study of 11 T. cruzi strains presented here included four parameters: growth kinetics, parasitemia curves, rate of macrophage infection, and serology to evaluate IgM, total IgG, IgG1, IgG2a, and IgG3. Sequencing of small subunit of ribosomal RNA (SSU rRNA)was performed and the T. cruzi strains were classified into three DTUs. When their growth in liver infusion tryptose medium was represented in curves, differences among the strains could be noted. The parasitemia profile varied among the strains from the TcI, TcII, and TcIII groups, and the 11 T. cruzi strains produced distinct parasitemia levels in infected BALB/c. The TcI group presented the highest rate of macrophage infection by amastigotes, followed by TcII and TcIII. Reactivity to immunoglobulins was observed in the TcI, TcII, and TcIII; all the animals infected with the different strains of T. cruzi showed anti-T. cruzi antibodies. The molecular study presented here resulted in the classification of the T. cruzi strains into the TcI (Bolivia, T lenti, Tm, SC90); TcII (Famema, SC96, SI8, Y); and TcIII (QMM3, QMM5, SI5) groups. These biological and molecular results from 11 T. cruzi strains clarified the factors involved in the biology of the parasite and its hosts. The collection of triatomine (vector) species, and the study of geographic distribution, as well as biological and molecular characterization of the parasite, will contribute to the reporting and surveillance measures in Brazilian states.

Lemnicola hungarica (Bacillariophyceae) and the new monoraphid diatom Lemnicola uniseriata sp. nov. (Bacillariophyceae) from South Korea

Many achnanthoid diatoms were transferred from Achnanthes or Achnanthidium to new genera based on characteristics observed via scanning electron microscopy (SEM). In this study, we isolated and cultured two Korean freshwater achnanthoid diatoms (Bacillariophyceae), studied their morphological characteristics using SEM, and assessed their molecular characteristics by analysing the nuclear small subunit (SSU) and ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) genes. We emended the description of Lemnicola hungarica with regard to the spindly pore opening of the hymen and the internally convex occlusions of elongated pores on the mantle. In addition, we demonstrated that the valves of L. hungarica are occasionally biseriate to uniseriate towards the valve edge. A morphological comparison of Lemnicola strains and species showed that biseriate rows of areolae are no longer an essential characteristic for allocation to Lemnicola. Based on the SEM and molecular analysis results, we also established a new, small-celled monoraphid species, Lemnicola uniseriata sp. nov., which has elongated areolae occluded by radiately structured hymenes and an elliptic valve shape. The uniseriate areolae of L. uniseriata showed that Lemnicola includes both uniseriate and biseriate striae. Additionally, the results of SSU and rbcL sequence analyses provided strong evidence to support the recognition of L. uniseriata as a new species.

Molecular and morphological characterisation of Aphelenchoides kheirii n. sp. (Nematoda: Aphelenchoididae) isolated from Pinus nigra in north-western Iran

SummaryAphelenchoides kheiriin. sp. was isolated during a survey of nematodes associated with bark samples of pine trees (Pinus nigra) in West Azerbaijan province, north-western Iran. The new species has a body length of 500 (448-520) μm in the female and 442 (402-480) μm in the male. The cuticle is weakly annulated with four lateral lines and the lip region is offset. The stylet is 10.4 (10-11) μm long with small basal swellings and the excretory pore located immediately posterior to base of metacorpus; hemizonid invisible. The post-vulval uterine sac length 37.7 (28-45) μm. Spicules are relatively short (18-20 μm in dorsal limb) and the end of the dorsal limb clearly curved ventrally like a hook. The male tail has usual three pairs of caudal papillae (2 + 2 + 2) and a well-developed mucron. The female tail is conical, terminating in a complicated step-like projection, usually with many tiny nodular protuberances. The new species belongs to the Group 2 category ofAphelenchoidesspeciessensuShahina in which ten known species among Groups 2 and 4sensuShahina, namely:A. arcticus,A. blastophthorus,A. fuchsi,A. parasaprophilus,A. paraxui,A. xui,A. dactylocercus,A. gynotylurus,A. iranicusandA. saprophilusare the most similar species. Phylogenetic analysis based on small subunit (SSU) and partial large subunit (LSU) sequences of rRNA supported the morphological results.

Morphological and molecular characterisation of three species of Paratylenchus Micoletzky, 1922 (Tylenchida: Paratylenchidae) from China, with a first description of the male P. rostrocaudatus

Summary In a recent survey of plant-parasitic nematodes associated with forest in China, three species of Paratylenchus, including P. aculentus, P. leptos and P. rostrocaudatus, were found. Paratylenchus leptos and P. rostrocaudatus are new records for China and the male of P. rostrocaudatus is described for the first time. Morphological descriptions, morphometrics and light microscopic photos are provided for these three species. Molecular characterisation of these species using ribosomal RNA (rRNA) D2-D3 expansion domains of large subunit (LSU D2-D3), internal transcribed spacer (ITS) and small subunit (SSU) sequences are also given. Except for the ITS sequence of P. aculentus, other sequences were amplified for the first time. Phylogenetic relationships within the genus Paratylenchus are discussed as inferred from the analyses of LSU D2-D3, ITS and SSU.

Arthothelium punctatum (Arthoniaceae, Arthoniales), A New Lichen Species from South Korea

A total of 121 species of lichens belonging to the genus Arthothelium have been described to date, most of which have been found in tropical regions. Here, we describe the discovery of a novel Arthothelium species for the first time in South Korea. Until now, Arthothelium ruanum was the only Arthothelium species reported in South Korea. Among the 113 specimens collected in this study, we identified A. ruanum and a putative new species, Arthothelium punctatum (J. S. Park & J.-S. Hur, sp. nov.). The diagnostic characters of A. punctatum are as follows: apothecia punctate, shortly elongate to branched, small, 0.1–0.2 mm wide, hypothecium hyaline to pale brown and obovate to broadly ellipsoid, muriform ascospores, 29.5–44.6 × 12.2–18.2 μm. The new species was found in Mt. Seokbyeong at an altitude of 790 m on smooth bark. Upon phylogenic analysis, the putative new species, A. punctatum, was separated from other Arthothelium species although the specimens analyzed were clustered with Arthoniaceae in phylogenetic trees based on both the mitochondrial small subunit (mtSSU) sequence and combined mtSSU and nuclear ribosomal large subunit sequences. Our data clearly indicate that this species is a new species belonging to the family Arthoniaceae. To elucidate the taxonomic characteristics of the new species, we provide morphological descriptions and a distribution map.

Taxonomic Revision of the Lichen Genera Pertusaria, Varicellaria, and Variolaria (Pertusariales, Ascomycota) in South Korea

The crustose lichen genus Pertusaria comprises over ca. 800 species worldwide. In total, 20 Pertusaria species were localized to the Mt. Sorak and Jeju-do in Korea. To date, information regarding the distribution of Pertusaria species in the South Korean peninsula is scarce. In this study, we collected Pertusaria species across South Korea and identified them based on morphological, chemical, and molecular characteristics. Of the 387 samples collected, we identified 24 taxa and 1 variety, of which 17 were previously recorded, and 6 taxa were newly found in South Korea (P. leioplaca, P. leucosora var. violascens, P. texana, P. thiospoda, P. thwaitesii, and P. xanthodes), 2 known species were transferred to Varicellaria (Varicellaria lactea and V. velata), one species was transferred to Variolaria as a new record (Variolaria multipunctoides) and one was a new species (P. jogyeensis J. S. Park & J.-S. Hur, sp. nov.). Characteristics of the newly discovered species, P. jogyeensis, are as follows: smooth to bumpy thallus, scattered to crowded poriform apothecia, blackish ostioles, definitely sunken, thin yellowish green rims around ostioles, 8-spored ascus, and the presence of perlatolic acid and thiophaninic acid (chlorinated xanthone). Phylogenetic studies on P. jogyeensis based on the mitochondrial small subunit sequence revealed proximity to P. flavicans and P. texana, and supported its classification as a new species within the genus Pertusaria. Additionally, we describe the chemical composition and morphology of all listed species in detail and provide an artificial key for identification.