Partial Large Subunit Research Articles

Glyphidohaptor safiensis n. sp. (Monogenea: Ancyrocephalidae) from the white-spotted rabbitfish Siganus canaliculatus (Park) (Perciformes: Siganidae) off Oman, with notes on its phylogenetic position within the Ancyrocephalidae Bychowsky & Nagibina, 1968 (sensu lato).

A new ancyrocephalid monogenean is described from the gills of wild White-spottedrabbitfish Siganus canaliculatus (Park) based on morphological and molecular analyses. Glyphidohaptor safiensis n. sp. can be distinguished from other members of the genus by the shape of the accessory piece of the male copulatory organ (MCO). Unlike its congeners, the rod-shaped accessory piece of G. safiensis n. sp. is distally broad and flattened. The MCO of G. safiensis n. sp. is curved, enclosed in a heavy sheath with a terminal flap. Partial large subunit (LSU), partial small subunit (SSU) and the partial SSU, entire internal transcribed spacer region 1 (ITS1) and partial 5.8S rDNA of the new species and two species of Tetrancistrum Goto & Kikuchi, 1917 from the same host and locality were sequenced and subjected to phylogenetic analysis. The LSU rDNA analysis grouped G. safiensis n. sp. with Tetrancistrum sp. from the gills of Siganus fuscescens Houttuyn from Australia, indicating a possible misidentification of the latter. Sequences of the SSU rDNA of the new species were most similar to those for Pseudohaliotrema sphincteroporus Yamaguti, 1953, demonstrating the close relatedness of the genera Pseudohaliotrema Yamaguti, 1953 and Glyphidohaptor Kritsky, Galli & Yang, 2007 within the Ancyrocephalidae. The comparison of the partial SSU (424 bp) and entire ITS1 and partial 5.8S rDNA (246 bp) sequences obtained for G. safiensis n. sp. with the only available sequence of another member of Glyphidohaptor Kritsky, Galli & Yang, 2007, G. pletocirra Paperna, 1972 (HE601931-HE601933) yielded on average 1.08% dissimilarity (a difference of 7 bases), with a p-distance of 0.010 ± 0.004%. This is the first record of a species of Glyphidohaptor from S. canaliculatus and from the Persian Gulf, the Gulf of Oman and the Arabian Sea.

Karlodinium elegans sp. nov. (Gymnodiniales, Dinophyceae), a novel species isolated from the East China Sea in a dinoflagellate bloom

On May 24–29, 2019, a harmful algal bloom occurred in Pingtan coastal areas, Fujian, southeast China, and caused mass mortality of cage-cultured fish. Two clonal cultures of an unknown naked dinoflagellate were set up from seawater samples taken during the bloom. The cultures were examined for morphological features, ultrastructure characters, photosynthetic pigments, the large subunit (LSU) of the rRNA gene, and the internal transcribed spacer (ITS) sequences, as well as acute toxicity analysis. The cell was unarmored, small-sized, and ovoid, and was characterized by elaborate striations on the epicone and hypocone. The nucleus was large, ellipsoid to oval or kidney-shaped, and centrally located in the cell. A long linear apical groove originated above the sulcus in the ventral epicone and extended to the dorsal side. An elongate, slit-like “ventral pore” was located on the left of the epicone, well away from the apical groove. The chloroplasts were yellowish brown, numerous, band like, and irregularly distributed in the cell periphery. Fucoxanthin was the main accessory pigment composition. Phylogeny topology reconstructed on partial LSU rDNA showed that the unknown dinoflagellate branched as a sister species to Karlodinium sp. (strain IFR981 & IFR797, from France) and Karlodinium corrugatum (strain KDGSO08, from Australia), with genetic divergences of 0.6% and 3.3%, respectively. Based on the morphological and molecular phylogenetic analysis, we describe the novel dinoflagellate as Karlodinium elegans sp. nov. A toxicity assay revealed that the clonal culture of K. elegans (strain PTB601) had no adverse effect on brine shrimp (Artemia salina) and marine medaka (Oryzias melastigma), indicating it may not be a toxic species.

First Report of Gaeumannomyces radicicola causing stalk rot on maize in Brazil.

Maize (Zea mays L.) is one of the most important commodities, and Brazil is the second-largest maize exporter country in the world. In April 2019, the period of the second crop maize (safrinha), it was observed black decayed lesions on roots and wilting of some maize plants, causing a "sudden death" in a commercial area in the west of Paraná state, Brazil (Figure 1A-C). Symptomatic root and stalk were collected, and tissues surface disinfected with 70% ethanol for 30 s, 1.5% NaOCl for 1 min and rinsed three times in sterile distilled water, slices of necrotic tissues were transferred to potato dextrose agar (PDA) medium and grown for 7 days at 27 ± 1ºC with a photoperiod of 12 h. Pure cultures were obtained through monosporic isolation. The fungal morphology is alike Gaeumannomyces radicicola, which is a synonym of Phialophora radicicola var. radicicola, Harpophora radicicola, P. zeicola, H. zeicola and G. graminis var. maydis (Hernández-Restrepo et al. 2016). Colonies on PDA showed flat, white to light gray at first (Fig. 1D), turning gray to black with age (Fig. 1E). Colony diameter approximately 5.2 cm on PDA in the dark after 7 days at 27ºC. Conidiophores with slightly thickened wall, mostly branched, varying in dimensions, with a range of 57.5-166.5 (avg. 128.7 μm) × 2.9-5.9 (avg. 4.2 μm) n = 25 (Fig. 1H-J). The conidia showed lunate-shaped with rounded ends, produced successively at the apex of phialide, 3.3-9.7 (avg. 6.6 μm) × 1.5-3.6 μm (avg. 2.5 μm), n = 100 (Fig. 1G-J). Morphological characteristics were comparable to the description of this specie (Cain 1952; Gams 2000; McKeen 1952). The total genomic DNA of a representative isolate, LEMIDPRZm 19-01 was extracted and the partial large subunit (28S nrDNA; LSU), internal transcribed spacer nrDNA including the intervening 5.8S nrDNA (ITS), and part of the largest subunit of the RNA polymerase II gene (RPB1) were amplified and sequenced, as following by Hernández-Restrepo et al. (2016) and Klaubauf et al. (2014). The primers to LSU - NL1 (O'Donnel, 1993) and LR5 (Vilgalys; Hester, 1990); ITS - ITS5 and ITS4 (White et al., 1990); and RPB1 - RPB1F and RPB1R (Klaubauf et al., 2014) were used in this study. The gene sequences of LSU (MT123866), ITS (MT114427), and RPB1 (MT123867) were deposited in GenBank and showed 99.67%, 99.75%, and 100% identity with type material G. radicicola CBS 296.53 (KM484962, KM484845, and KM485061). A multi-locus phylogenetic analysis based on Bayesian Inference showed the isolate LEMIDPRZm 19-01 in the G. radicicola clade (Fig. 2). To confirm pathogenicity, ex vivo assays were performed with mycelial PDA discs of 5 mm from a 7-day-old culture using detached roots (adapted method by Degani et al., 2019), on wounded and unwounded stalk and leaves, each treatment consisted of five replications. PDA discs without fungal were used in negative tissue controls. Pathogenicity tests were also conducted in vivo, two experiments performed: i) the stalk tissue was inoculated by sterilized toothpick grown on PDA with fungal mycelium and the leaves inoculated as ex vivo assay, and toothpick without fungal mycelium was used to stalk negative control, whereas PDA discs without fungal were used in the tested leaves; ii) 6 mycelial PDA discs/500 mL were placed on potato dextrose broth (PDB) media and it remained in agitation for 10 days to obtain a mycelial suspension. Subsequently, the mycelial was crushed to soil infestation, and 50 mL from this suspension were dropped in each 2 L maize pot with soil sterilization 10 days after emergence. Maize pots with soil sterilization without mycelium fungal were used as negative controls. Four replications (maize pots), for each treatment, were used in both tests. Experiments were repeated twice. In the ex vivo assay, all inoculated tissues with and without wounds showed necrotic lesions (Fig. 1K-N). In the first in vivo assay, stalk rot symptoms, including wilting of the inoculated plants causing premature plant death, were observed within 6 days (Fig. 1O-Q). In the second in vivo assay, inoculated plants had inferior growth than compared with plant control. Sixty days after inoculation, the plants were removed from the pots and it was observed a roots degeneration with symptoms of necrosis (Fig. 1R-U). No symptoms were detected in the control treatments and the pathogen was re-isolated from symptomatic tissues confirming Koch's postulate for all assays. So far, to our knowledge, the pathogen distribution was reported solely in the west area of Paraná state, but it may become a potential threat to Brazilian maize production. Further monitoring is necessary to better understand the epidemiology of this pathogen to address a strategy for disease control. The pathogen has already been detected in Canada, South Africa, and China. To our knowledge, this is the first report of G. radicicola in Brazil, as well as in South America.

A New Species of Thelonectria and a New Record of Cephalotrichum hinnuleum from Gunwi and Ulleungdo in Korea

Three fungal strains belonging to the class Sordariomycetes were isolated from soils collected from Gyeongsangbuk-do in Korea. They were identified as Cephalotrichum hinnuleum (UD CT 1-3-3 and KNU-19GWF1) and Thelonectria chlamydospora sp. nov. (UD ST 1-2-1). T. chlamydospora sp. nov. was morphologically identical to T. truncata, but its specific macroconidial dimensions, lower number of septations, and chlamydospore diameter render it distinct from the strains of the genus Thelonectria. The strains UD CT 1-3-3 and KNU-19GWF1 were developed flat, velvety to felty, and golden gray to brown-gray after 14 days of incubation at 25 °C on PDA. These strains were produced polyblastic conidiogenous cells and conidia were pale brown to brown, smooth, thin-walled, subglobose to ellipsoidal, arranged in chains, and the diameters of 6.7–9.0 × 3.7–5.1 μm. The strains were also confirmed by using the multi-locus genes using internal transcribed spacer (ITS) regions, partial large subunit (LSU), translation elongation factor 1α (TEF1-α), β-tubulin (TUB2), and actin (ACT) genes. This is the discovery of T. chlamydospora sp. nov. and Cephalotrichum hinnuleum, a new record from Korea.

A 28S rDNA‐based phylogeny of the nematode family Acuariidae (Spirurida) parasitic in vertebrates

AbstractThe aim of this study was to investigate the phylogenetic relationships of nematodes of the family Acuariidae using partial large subunit nuclear ribosomal DNA (28S) sequences of 15 genera represented by 18 species. The results confirmed the monophyly of the family Acuariidae and supported its close relationship with nematodes of the family Cystidicolidae. Two major clades were revealed within Acuariidae, one represented by nematodes of the subfamily Schistorophinae and another composed of members of the subfamilies Acuariinae and Seuratiinae. The collarette was shown to be a structure that arose several times within the clade Acuariinae–Seuratiinae. As a result, we suggest the suppression of the subfamily Seuratiinae and inclusion of its genera within the subfamily Acuariinae. Morphological characters, host ranges and life cycles of the taxa included in the analyses are discussed as well as the possible relationships of remaining acuariid genera within the revealed subclades. Additionally, we propose an amended generic diagnosis of Syncuaria Gilbert, 1927 to accommodate Syncuaria sagittata (Rudolphi, 1809) n. comb. and Syncuaria longevaginata (Molin, 1860) Skrjabin, Sobolev, & Ivashkin, 1965. New host and geographic records are also presented.

<p><strong>A new species of <em>Podosphaera</em> sect. <em>Sphaerotheca</em> subsect. <em>Magnicellulatae</em> from India and a key to all species reported on <em>Fabaceae</em></strong></p>

Podosphaera thermopsidicola, a new sexual morph form of powdery mildew was discovered on living leaves of Thermopsis barbata Benth. (Fabaceae). It is described, illustrated and compared with other Podosphaera species reported on Fabaceae. The phylogeny of this species has been inferred from partial nuclear ribosomal 28S large subunit (LSU) and internal transcribed spacer (ITS) rDNA sequence data. On the basis of LSU, P. thermopsidicola represents characteristic feature of Podosphaera s. str. Phylogenetically, P. thermopsidicola is closer to P. erigerontis-canadensis but differs in having larger chasmothecia, longer appendages, thick-walled asci, smaller oculi and smaller ascospores.

Morphology, ultrastructure, and molecular phylogeny of the unarmoured dinoflagellate Kirithra sigma sp. nov. (Ceratoperidiniaceae, Dinophyceae)

ABSTRACT A Gymnodinium-like dinoflagellate, Kirithra sigma sp. nov., was isolated from Jiaozhou Bay, China, and the southern coast of Korea, and examined by light, scanning, and transmission electron microscopy. Pigment composition and nuclear-encoded partial large subunit ribosomal RNA (LSU rRNA) gene were determined. Cells were oval-shaped (26.4‒36.4 μm long and 17.7‒27.2 μm wide), with a conical epicone and hemispherical hypocone. Its cingulum and sulcus were deep and wide. The cingulum encircled the cell completely, its ends slightly overlapping and displaced one to two cingular widths; the sulcus and sulcal extension formed a sigmoid curve. The anterior sulcal extension contacted the apical structure complex (ASC). The ASC formed a complete circle, consisting of three parallel series of vesicles, and the amphiesmal vesicles within ASC were arranged with one central vesicle (sometimes two) encircled by approximately three rows of polygonal amphiesmal vesicles. Polygonal amphiesmal vesicles of different sizes covered the whole cell surface. The subspherical nucleus was located in the hypocone. Pigment analysis showed that peridinin was the main accessory pigment. The present species is morphologically similar to Kirithra asteri but differs in numbers and rows of amphiesmal vesicles within the ASC, absence of ventral pore, and a sigmoid curve formed by the sulcus and sulcal extension. The partial LSU rRNA gene sequence (c. 1400 bases) of K. sigma differed by c. 5.3% from that of K. asteri. Molecular phylogeny revealed that K. sigma formed an independent sister clade to K. asteri, indicating they are congeneric but different species.

Stemmatostoma cribbi n. sp. (Digenea: Cryptogonimidae) from Freshwater Fishes in the Wet Tropics Bioregion of Queensland, Australia.

A survey of the parasite fauna of freshwater fishes from the Wet Tropics Bioregion in Queensland, Australia, revealed the presence of a new species of Stemmatostoma Cribb, 1986 (Digenea: Cryptogonimidae). Stemmatostoma cribbi n. sp. is described from the intestine and pyloric caeca of 2 species of grunter (Terapontidae), Hephaestus fuliginosus (Macleay) and Hephaestus tulliensis (De Vis), and the Jungle perch (Kuhliidae), Kuhlia rupestris (Lacepède), collected from the Barron and Mulgrave-Russell River drainage divisions in tropical north Queensland, Australia. Stemmatostoma cribbi is primarily distinguished morphologically from the type and only other species in the genus, Stemmatostoma pearsoni Cribb, 1986, in having consistently fewer oral spines (14 in S. cribbi vs. 16 in S. pearsoni). Alignment of novel molecular data for S. cribbi and S. pearsoni revealed that they differ genetically by 26 nucleotides (2.1%) over the 1,258 bp partial large subunit (LSU) region, 1 nucleotide (0.8%) over the 121 bp partial 5.8S region, and 23 nucleotides (7.2%) over the entire 318 bp ITS2 rDNA region. Bayesian inference and maximum likelihood phylogenetic analyses of the partial LSU region for the species of Stemmatostoma sequenced here were used to explore the relationships of these species to other cryptogonimid species reported from freshwater ecosystems.

Morphology, DNA Phylogeny, and Pathogenicity of Wilsonomyces carpophilus Isolate Causing Shot-Hole Disease of Prunus divaricata and Prunus armeniaca in Wild-Fruit Forest of Western Tianshan Mountains, China

Prunus divaricata and Prunus armeniaca are important wild fruit trees that grow in part of the Western Tianshan Mountains in Central Asia, and they have been listed as endangered species in China. Shot-hole disease of stone fruits has become a major threat in the wild-fruit forest of the Western Tianshan Mountains. Twenty-five isolates were selected from diseased P. divaricata and P. armeniaca. According to the morphological characteristics of the culture, the 25 isolates were divided into eight morphological groups. Conidia were spindle-shaped, with ovate apical cells and truncated basal cells, with the majority of conidia comprising 3–4 septa, and the conidia had the same shape and color in morphological groups. Based on morphological and cultural characteristics and multilocus analysis using the internal transcribed spacer (ITS) region, partial large subunit (LSU) nuclear ribosomal RNA (nrRNA) gene, and the translation elongation factor 1-alpha (tef1) gene, the fungus was identified as Wilsonomyces carpophilus. The 25 W. carpophilus isolates had high genetic diversity in phylogenetic analysis, and the morphological groups did not correspond to phylogenetic groups. The pathogenicity of all W. carpophilus isolates was confirmed by inoculating healthy P. divaricata and P. armeniaca leaves and fruits. The pathogen was re-isolated from all inoculated tissues, thereby fulfilling Koch’s postulates. There were no significant differences in the pathogenicity of different isolates inoculated on P. armeniaca and P. divaricata leaves (p > 0.05). On fruit, G053 7m3 and G052 5m2 showed significant differences in inoculation on P. armeniaca, and G010 5m2 showed extremely significant differences with G004 7m2 and G004 5m2 on P. divaricata (p < 0.05). This is the first report on shot-hole disease of P. armeniaca (wild apricot) leaves and P. divaricata induced by W. carpophilus in China.

Isolation and identification of L-asparaginase-producing endophytic fungi from the Asteraceae family plant species of Iran.

L-asparaginase is an important anticancer enzyme that is used in the first line treatment of acute lymphoblastic leukemia. This study was conducted to isolate L-asparaginase-producing endophytic fungi from medicinal plants of family Asteraceae. Seven healthy medicinal plants from family Asteraceae were selected for the isolation of endophytic fungi using standard surface sterilization techniques. A total of 837 isolates belonging to 84 species were comprised of the stem (55.6%), leaf (31.1%), root (10.6%) and flower (2.7%). Initial screening of L-asparaginase-producing endophytes was performed by qualitative plate assay on modified Czapex dox’s agar medium. L-asparaginase activity of fungal endophytes was quantified by the nesslerization method. Identification of endophytic fungi was performed using both morphological characteristics and phylogenetic analyses of DNA sequence data including ribosomal DNA regions of ITS (Internal transcribed spacer) and LSU (partial large subunit rDNA), TEF1 (Translation Elongation Factor) and TUB (β-tubulin). Of the 84 isolates, 38 were able to produce L-asparaginase and their L-asparaginase activities were between 0.019 and 0.492 unit/mL with Fusarium proliferatum being the most potent. L-asparaginase-producing endophytes were identified as species of Plectosphaerella, Fusarium, Stemphylium, Septoria, Alternaria, Didymella, Phoma, Chaetosphaeronema, Sarocladium, Nemania, Epicoccum, Ulocladium and Cladosporium. This study showed that endophytic fungi from Asteraceae members have a high L-asparaginase-producing potential and they can be used as an alternative source for production of anticancer enzymes.

<p><strong><em>Strelitziana sarbhoyi </em></strong><strong>sp. nov.<em> </em>(Strelitzianaceae, Chaetothyriales), from North-Western Himalayas, India, described based on morphology and molecular phylogeny</strong></p>

Strelitziana sarbhoyi is established as a new species to accommodate a phylloplane fungus isolated from Mallotus philippensis collected from Kangra region of North-Western Himalayas, Himachal Pradesh. The identity of the fungus is confirmed based on the asexual-morphs, cultural characteristics and phylogenetic analyses of the internal transcribed spacer (ITS) rDNA and partial nuclear ribosomal 28S large subunit (LSU) sequence data. The placement of S. sarbhoyi in the phylogenetic tree was determined based on DNA sequences from authenticated isolates of Strelitziana. Strelitziana sarbhoyi shows nearly 94% similarity with other known species of Strelitziana. Area description is provided for the proposed taxon along with microscopic images, and a phylogenetic tree. This is probably the first report of Strelitziana from India. Holotype specimen (dried voucher culture) is deposited in the Ajrekar Mycological Herbarium (AMH), and an ex-type culture is deposited in National Fungal Culture Collection of India (NFCCI).

<p><strong><em>Kamalia indica,</em></strong><strong> a new genus and species belonging to Diaporthales (Ascomycota, Fungi) described based on morphology and molecular phylogeny</strong></p>

A fungal taxa isolated from leaf spots of Mallotus philippensis from Kangra district of North-Western Himalayan region of India is established as a new genus based on morphological characters of asexual-morphs, cultural characteristics and phylogenetic analyses of the partial nuclear ribosomal 28S large subunit (LSU) and internal transcribed spacer (ITS) rDNA sequence data. The generic placement of the genus has been determined based on DNA sequences from authenticated isolates. The present taxon has turned out to be distinct, showing nearly 90% identity with other known genera in Diaporthales based on nrDNA internal transcribed spacer region. The morphological description is provided for the new taxa and compared with the similar taxa belonging to the order Diaporthales. The culture was found to show heavy sporulation in all kind of media. The type specimen and ex type culture have been deposited in the Ajrekar Mycological Herbarium (AMH) and National Fungal Culture Collection of India (NFCCI-WDCM 932), respectively.

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.

Pseudoplagiostoma myracrodruonis (Pseudoplagiostomataceae, Diaporthales): a new endophytic species from Brazil

Two initially unidentifiable isolates were found when endophytic fungi from healthy leaves of Myracrodruon urundeuva (Anacardiaceae) were being investigated in Brazil. Based on phylogenetic analyses using internal transcribed spacer (ITS) and partial large subunit (LSU) of ribosomal DNA, RNA polymerase II (RPB2), translation elongation factor EF-1 alpha (TEF1), and tubulin (TUB2), the isolates were identified as members of the family Pseudoplagiostomataceae (Diaporthales). In this paper, we introduce a new species, Pseudoplagiostoma myracrodruonis, thus accepting eight species in Pseudoplagiostoma (Pseudoplagiostomataceae). The new species is proposed to accommodate endophytes characterized by a coelomycetous asexual morph, pycnidial conidiomata with lageniform to ampulliform and hyaline to pale brown phialidic conidiogenous cells that produce unicellular, ellipsoid to oblong-cylindrical, hyaline, and guttulate conidia. In the current phylogenetic analysis, P. myracrodruonis is placed in a well-supported clade in Pseudoplagiostoma (Pseudoplagiostomataceae). A discussion about the relationship of Pseudoplagiostoma species is included in the paper.

Identification and Characterization of Neofabraea kienholzii and Phlyctema vagabunda Causing Leaf and Shoot Lesions of Olive in California.

California produces over 95% of the olives grown in the United States. In 2017, California's total bearing acreage for olives was 14,570 hectares producing 192,000 tons of olives valued at $186.6 million. During the early spring of 2016, unusual leaf and shoot lesions were detected in olive trees from superhigh-density orchards in the Northern San Joaquin and Sacramento valleys of California. Affected trees displayed numerous leaf and shoot lesions developing at wounds created by mechanical harvesters. The 'Arbosana' cultivar was highly affected by the disease, whereas the disease was sporadic in 'Arbequina' and not found in 'Koroneiki' cultivar. Two fungal species, Neofabraea kienholzii and Phlyctema vagabunda, were found to be consistently associated with the disease, and Koch's postulates were completed. Species identity was confirmed by morphology and molecular data of the partial large subunit rDNA, the internal transcribed spacer region, and partial beta-tubulin region. The disease signs and symptoms are described and illustrated.

Polyphasic Discrimination of Trichophyton tonsurans and T. equinum from Humans and Horses.

The anthropophilic dermatophyte Trichophyton tonsurans and its zoophilic counterpart T. equinum are phylogenetically closely related. The barcoding marker rDNA internal transcribed spacer (ITS) shows limited variation between these two species. In the current study, we combined molecular approaches with phenotypic data to determine the species boundaries between T. tonsurans (n = 52) and T. equinum (n = 15) strains originating from humans (n = 40), horses (n = 26), and a mouse (n = 1). Culture characteristics and physiology on Trichophyton agar media 1 and 5 were evaluated. Multi-locus sequencing involving ITS, partial large rDNA subunit (LSU), β-tubulin (TUB), 60S ribosomal protein (RPB), and translation elongation factor-3 (TEF3) genes, and the mating-type (MAT) locus was performed. Amplified fragment length polymorphism data were added. None of the test results showed complete mutual correspondence. With the exception of strains from New Zealand, strains of equine origin required niacin for growth, whereas most strains from human origin did not show this dependence. It is concluded that T. tonsurans and T. equinum incompletely diverged from a common lineage relatively recently. MAT1-1 and MAT1-2 are the main distinguishing genes between the two species.

Ectophoma iranica sp. nov. and new hosts and records of Allophoma spp. in Iran

AbstractA survey was conducted to identify fungi associated with leaf diseases of ornamental herbs, including Christ plant, dracaena, rose geranium, rose periwinkle and treasure flower, in Ahvaz, southwestern Iran. Twenty‐five symptomatic herbs were collected and studied, which led to the isolation of seven Phoma‐like strains belonging to four taxa. These fungi were characterised based on DNA sequence data for the partial large subunit 28S nrDNA (LSU‐D1/D2), the internal transcribed spacer 1 and 2 and 5.8S nrDNA (ITS) and part of the β‐tubulin (tub2) gene regions. A multi‐locus‐based phylogeny, in combination with morphology, allowed for the identification of Allophoma hayatii, A. labilis, A. tropica and a novel species of the genus Ectophoma. Ectophoma iranica sp. nov. is morphologically and genetically distinctive from previously described species. In pathogenicity tests, the two strains of E. iranica (CBS 144681 and IRAN 3354C) caused leaf spot symptoms on leaves of Catharanthus roseus. New host plant species and records for A. hayatii, A. labilis and A. tropica are reported, plus further emended characterization of A. hayatii.

Phylogenetic relationships of Phaeochorella parinarii and recognition of a new family, Phaeochorellaceae (Diaporthales)

ABSTRACTSpecies of Phaeochorella are biotrophic leaf parasites with a tropical distribution, traditionally accepted in the family Phyllachoraceae, Phyllachorales in classifications based on morphological characters. Phylogenetic evidence presented here resolves the relationship of Phaeochorella within the Sordariomycetes, based on a multilocus analysis of partial nuc rDNA large subunit (28S) and internal transcribed spacers (ITS1-5.8S-ITS2 = ITS), the DNA-directed RNA polymerase II second largest subunit (RPB2), and the translation elongation factor 1-α (TEF1-α) gene. Phylogenetic analyses indicate that Phaeochorella belongs to the Diaporthales rather than the Phyllachorales. Phaeochorella parinarii, the type species of the genus, present on native hosts from the Brazilian Cerrado, forms a unique clade with a species of Phaeoappendicospora with high support. Thus, a new family, Phaeochorellaceae, Diaporthales, including both genera, is herein proposed. With the exception of P. parinarii and P. zonata, all other species in Phaeochorella (P. artocarpi, P. ciliata, P. machaerii) were excluded from the genus.

Genetic diversity, and description of a new dagger nematode, Xiphinema afratakhtehnsis sp. nov., (Dorylaimida: Longidoridae) in natural forests of southeastern Gorgan, northern Iran.

The most prevalent dagger nematode recovered from rhizospheric soil samples of forest trees in the Afrātakhteh region of Golestan province (Iran) was Xiphinema afratakhtehnsis sp. nov. and it is described and illustrated with integrative approaches using both morphological and molecular criteria. It belongs to the morphospecies group 6 of the intragenic historical grouping of Xiphinema non-americanum species. The new species is characterized by females with 3.3–4.9 mm sized body, lip region separated from the rest of body by a depression, anteriorly expanded, 16–18 μm wide, vulva located at 47.2–58.5%, odontostyle 155–173 μm and odontophore 89–107 μm long, female genital system composed of two equally developed branches, the tubular part of each having spines, short symmetrically rounded female tail to symmetrically rounded with a small mucro-like projection at the end in a few females, rare males (n = 1 out of 74 females) with 83 μm long dorylaimoid spicules and four juvenile developmental stages. The third-stage juveniles (J3) have a characteristic tail shape (short, symmetrically conical with a club-shaped long mucro) demarcating the species, and being typologically useful for its separation from closely similar species (except X. cohni, with currently no data on its juvenile stages) viz. X. adenohystherum, X. iranicum, X. mazandaranense, X. nuragicum, X. pyrenaicum, X. robbinsi, X. sphaerocephalum and X. zagrosense. Molecular phylogenetic studies using genomic (partial large subunit and internal transcribed spacer 1 ribosomal RNA genes: D2-D3 and ITS1 rDNA) and mitochondrial cytochrome c oxidase subunit I gene (COI mtDNA) revealed the new species forming a unique lineage in all reconstructed trees using Bayesian inference (BI) and maximum likelihood (ML) methods. The sequenced isolates of the new species formed a monophyletic group in the D2-D3 tree. The sequenced isolates of the new species for their COI mtDNA formed four subclades in COI mtDNA phylogeny, and four haplotypes in corresponding analysis.

Morphology, ultrastructure and molecular phylogeny of cyst-producing Caladoa arcachonensis gen. et sp. nov. (Peridiniales, Dinophyceae) from France and Indonesia

ABSTRACTThe dinoflagellate order Peridiniales encompasses several well circumscribed families. However, the family level of some genera, such as Bysmatrum and Vulcanodinium, has remained elusive for many years. Four Peridinium-like strains were established from the Atlantic coast of France and North Sulawesi, Indonesia through cyst germination or isolation of single cells. The cyst-theca relationship was established on specimens from the French Atlantic. Their morphologies were examined using light, scanning and transmission electron microscopy. The cells were characterized by a much larger epitheca relative to the hypotheca, a large anterior sulcal (Sa) plate deeply intruding the epitheca and a small first anterior intercalary plate. The plate formula was identified as Po, cp, X, 4′, 3a, 7′′, 6C, 5S, 5′′′, 2′′′′, shared by Apocalathium, Chimonodinium, Fusiperidinium and Scrippsiella of the family Thoracosphaeraceae but the configuration of Sa plate and anterior intercalary plates is different. Transmission electron microscopy showed that the eyespot was located within a chloroplast comprising two rows of lipid globules and thus belongs to type A. All four strains were classified within a new genus Caladoa as C. arcachonensis gen. et sp. nov. Small subunit ribosomal DNA (SSU rDNA), partial large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer ribosomal DNA (ITS rDNA) sequences were obtained from all strains. Genetic distance based on ITS rDNA sequences between French and Indonesian strains reached 0.17, suggesting cryptic speciation in C. arcachonensis. The maximum likelihood and Bayesian inference analysis based on concatenated data from SSU and LSU rDNA sequences revealed that Caladoa is monophyletic and closest to Bysmatrum. Our results supported that Caladoa and Bysmatrum are members of the order Peridiniales but their family level remains to be determined. Our results also support that Vulcanodinium is closest to the family Peridiniaceae.