Molecular phylogeny and morphology of <Emphasis Type="Italic">Ophiocordyceps unituberculata</Emphasis> sp. nov. (Ophiocordycipitaceae), a pathogen of caterpillars (Noctuidae,Lepidoptera) from Yunnan,China

Ophiocordyceps unituberculata, a new insect pathogenic fungus from southwestern China, is described using molecular phylogenetic and morphological data. This fungus differs from other Ophiocordyceps species by its enormously long monophialidic conidiogenous cells and a large periclinal protuberance often growing near the apex of conidiogenous cells, single conidia (lanceolate to fusiform) embedded in mucous sheath, and much larger conidial dimensions. Four-locus (nrSSU, nrLSU, tef-1α and rpb1) and ITS data phylogenetic analyses show that O. unituberculata belongs to the Hirsutella nodulosa clade within the genus Ophiocordyceps of Ophiocordycipitaceae and is a separate clade from other allied species. Molecular phylogeny and morphology both strongly support the distinctiveness of this taxon. The interspecific relationships in the H. nodulosa clade are discussed.     

<Emphasis Type="Italic">Melampsora salicis-bakko</Emphasis>, a new species on willows in Japan evidenced by morphological and molecular phylogenetic analyses

Through the morphological and molecular examinations of Melampsora species on willows, we clarified the taxonomic identity of the rust specimens on Salix bakko, S. hultenii and S. leucopithecia from Japan and described the following rust fungus as a new species, Melampsora salicis-bakko. This rust fungus resembled M. caprearum in morphology of teliospores, but it differed from M. caprearum mainly in the density of spines on the urediniospores. Molecular phylogenetic analyses using the rDNA ITS region (complete ITS1, 5.8S rRNA gene and ITS2) revealed that M. salicis-bakko was monophyletic, and that this rust fungus was distinct from other Melampsora species, including M. caprearum.     

Detection of <Emphasis Type="Italic">Rickettsia</Emphasis> spp. in ticks parasitizing toads (<Emphasis Type="Italic">Rhinella marina</Emphasis>) in the northern Brazilian Amazon

This study evaluated rickettsial infection in ticks collected on toads from the northern Brazilian Amazon (Amapá state), where to our knowledge there are neither records of ticks from amphibians nor rickettsial infections in ticks. During 2016–2017, a total of 22 free-living toads were captured and identified as Rhinella marina. Overall, 12 (54.5%) toads were parasitized by a total of 97 ticks (6 males, 39 females, 31 nymphs, 21 larvae), giving a mean intensity of 8.1 ticks per infested toad. Two tick species were morphologically identified: Amblyomma rotundatum Koch (31 females, 14 nymphs), and Amblyomma dissimile Koch (6 males, 8 females, 17 nymphs). The 21 larvae were morphologically denoted as Amblyomma sp. Five toads were co-infested by A. rotundatum and A. dissimile. Morphological identifications were confirmed by nucleotide sequencing of fragments of the mitochondrial genes 16S rDNA, 12S rDNA and/or COX1. A total of 54 ticks were analyzed for the presence of rickettsial DNA. Eleven (9 females and 2 nymphs) out of 14 A. rotundatum ticks contained Rickettsia bellii. None of the 25 specimens of A. dissimile (6 males, 6 females, 13 nymphs) contained amplifiable rickettsial DNA. From 15 Amblyomma sp. larvae, a pool of 10 individuals contained Rickettsia sp. strain Colombianensi. Sequencing of the 16S rDNA amplicon derived from the positive pool yielded a sequence of A. dissimile. We detected Rickettsia sp. strain Colombianensi for the first time in Brazil. Prior records of this agent were restricted to Colombia and Honduras. In addition, we report the presence of A. rotundatum for the first time in the state of Amapá, where the only other record of A. dissimile was registered over 20 years ago.     

Taxonomic revisions in the Microstromatales: two new yeast species,two new genera,and validation of <Emphasis Type="Italic">Jaminaea</Emphasis> and two <Emphasis Type="Italic">Sympodiomycopsis</Emphasis> species

Environmental sampling yielded two yeast species belonging to Microstromatales (Exobasidiomycetes, Ustilaginomycotina). The first species was collected from a leaf phylloplane infected by the rust fungus Coleosporium plumeriae, and represents a new species in the genus Jaminaea, for which the name Jaminaea rosea sp. nov. is proposed. The second species was isolated from air on 50% glucose media and is most similar to Microstroma phylloplanum. However, our phylogenetic analyses reveal that species currently placed in Microstroma are not monophyletic, and M. phylloplanum, M. juglandis and M. albiziae are not related to the type species of this genus, M. album. Thus, Pseudomicrostroma gen. nov. is proposed to accommodate the following species: P. glucosiphilum sp. nov., P. phylloplanum comb. nov. and P. juglandis comb. nov. We also propose Parajaminaea gen. nov. to accommodate P. albizii comb. nov. and P. phylloscopi sp. nov. based on phylogenetic analyses that show these are not congeneric with Jaminaea or Microstroma. In addition, we validate the genus Jaminaea, its respective species and two species of Sympodiomycopsis and provide a new combination, Microstroma bacarum comb. nov., for the anamorphic yeast Rhodotorula bacarum. Our results illustrate non-monophyly of Quambalariaceae and Microstromataceae as currently circumscribed. Taxonomy of Microstroma and the Microstromataceae is reviewed and discussed. Finally, analyses of all available small subunit rDNA sequences for Jaminaea species show that J. angkorensis is the only known species that possess a group I intron in this locus, once considered a potential feature indicating the basal placement of this genus in Microstromatales.     

Two new and one known species of <Emphasis Type="Italic">Tergestia</Emphasis> Stossich, 1899 (Trematoda: Fellodistomidae) with novel molecular characterisation for the genus

Combined morphological and molecular analyses are employed to characterise three species of Tergestia Stossich, 1899 (Digenea: Fellodistomidae) from fishes of Moreton Bay, Queensland, Australia. Tergestia clonacantha Manter, 1963 is reported here for the first time from the halfbeak (Beloniformes: Hemiramphidae) species Arrhamphus sclerolepis krefftii (Steindachner), Hyporhamphus australis (Steindachner), H. quoyi (Valenciennes) and H. regularis ardelio (Whitley). Two new species, both infecting trevally (Perciformes: Carangidae) species, are described: T. maryae n. sp. from Alepes apercna Grant and T. henryi n. sp. from Pantolabus radiatus (MacLeay). Complete ITS2 and partial 28S ribosomal DNA data were generated for each of the new taxa. The three species differ from each other by 47–58 base pairs (bp) in the ITS2 rDNA region. Phylogenetic analysis of 28S rDNA supports Tergestia as a reliable generic concept, with our analyses showing that some species of the genus form a well-supported clade to the exclusion of all other fellodistomids for which sequence data are available.     

<Emphasis Type="Italic">Pattersoniomyces tillandsiae</Emphasis> gen. et comb. nov.: linking sexual and asexual morphs of the only known smut fungus associated with Bromeliaceae

The enigmatic species Ustilago tillandsiae is the only known smut fungus associated with Bromeliaceae. Its generic position is evaluated by morphological, physiological, and molecular phylogenetic analyses using large subunit rDNA sequences. Phylogenetic analyses resolved U. tillandsiae as a member of the Ustilaginales in a sister relationship to the lineage containing Tranzscheliella species. However, U. tillandsiae differs from Tranzscheliella species by the development of sori in flowers, a different structure of sori and a different type of spore ornamentation. Consequently, a new genus Pattersoniomyces is described to accommodate U. tillandsiae. The new combination Pattersoniomyces tillandsiae is substantiated. In the sexual stage (teleomorph), this species infects bromeliads: Tillandsia flabellata, Tillandsia leiboldiana, and Tillandsia sp. in Central America between southern Mexico and Costa Rica. The yeast stage (anamorph) of P. tillandsiae was found associated with the phylloplane of Canistrum improcerum and in water tanks (phytotelmata) of Vriesea minarum, two bromeliads occurring in northeast and southeast Brazil, respectively. The link between the teleomorph and anamorphic strains is supported by identical sequences of the D1/D2 domains of the large subunit rDNA. Pattersoniomyces represents the tenth endemic smut genus to the Americas, but the only one that occurs in both North and South America, being a truly neotropical genus. The host plant families of Ustilaginales are extended to the Bromeliaceae. As far as we know, Pattersoniomyces represents the single event of a host jump from Cyperaceae or Poaceae to Bromeliaceae, apparently without further species radiation on multiple bromeliad species and genera growing in South America.     

The genus <Emphasis Type="Italic">Fusariella</Emphasis>

The genus Fusariella, typified by F. atrovirens, is characterised by semi- to macronematous, mononematous conidiophores, with cylindrical, subulate or lageniform phialidic conidiogenous cells that produce catenate, septate, curved to straight, subhyaline to brown conidia. During a survey of hyaline-spored hyphomycetes from karst areas in Thailand, we collected a new species of Fusariella with curved conidia and introduce it in this paper as Fusariella curvata sp. nov. In addition, all hitherto described species of Fusariella are reviewed. The result of phylogenetic analyses, based on combined SSU, LSU, TEF and RPB2 sequence data, indicates that the genus belongs in the family Bionectriaceae (Hypocreales, Sordariomycetes).     

Molecular marker to identify the fungus gnat, <Emphasis Type="Italic">Bradysia</Emphasis> sp. (Diptera: Sciaridae), a new pest of Welsh onion and carrot in Japan

A new pest of Welsh onion and carrot, the dark-winged fungus gnat, Bradysia sp., was found in the northern region of Saitama Prefecture, Japan. DNA sequences of mitochondrial cytochrome c oxidase subunit I revealed that Bradysia sp. is identical to the Chinese chive gnat, Bradysia odoriphaga Yang and Zhang. This suggests that Bradysia sp. may be an invasive species that originates from continental East Asia. Direct sequencing analysis and PCR using a species-specific primer were found to be useful tools for discriminating Bradysia sp. from other native Japanese fungus gnats, such as Bradysia impatiens (Johannsen), Pnyxia scabiei (Hopkins), and Lycoriella ingenua (Dufour).     

New species of the genus <Emphasis Type="Italic">Spathius</Emphasis> Nees (Hymenoptera,Braconidae) from Japan and neighboring territories

Eight new species of the genus Spathius Nees from Japan and the Russian Far East are described and illustrated: S. enigmatus sp. n., S. ishigakus sp. n., S. konishii sp. n., S. multigranulatus sp. n., S. omotodakus sp. n., S. pseudaspersus sp. n., S. pumilio sp. n., and S. tsushimus sp. n.     

<Emphasis Type="Italic">Lepotrema</Emphasis> Ozaki, 1932 (Lepocreadiidae: Digenea) from Indo-Pacific fishes,with the description of eight new species,characterised by morphometric and molecular features

We review species of the genus Lepotrema Ozaki, 1932 from marine fishes in the Indo-West Pacific. Prior to the present study six species were recognised. Here we propose eight new species on the basis of combined morphological and molecular analysis: Lepotrema acanthochromidis n. sp. ex Acanthochromis polyacanthus from the Great Barrier Reef (GBR); Lepotrema hemitaurichthydis n. sp. ex Hemitaurichthys polylepis and H. thompsoni from Palau and French Polynesia; Lepotrema melichthydis n. sp. ex Melichthys vidua from Palau and the GBR; Lepotrema amansis n. sp. ex Amanses scopas from the GBR; Lepotrema cirripectis n. sp. ex Cirripectes filamentosus, C. chelomatus and C. stigmaticus from the GBR; Lepotrema justinei n. sp. ex Sufflamen fraenatum from New Caledonia; Lepotrema moretonense n. sp. ex Prionurus microlepidotus, P. maculatus and Selenotoca multifasciata from Moreton Bay; and Lepotrema amblyglyphidodonis n. sp. ex Amblyglyphidodon curacao and Amphipron akyndynos from the GBR. We also report new host records and provide novel molecular data for two known species: Lepotrema adlardi Bray, Cribb & Barker, 1993 and Lepotrema monile Bray & Cribb, 1998. Two new combinations are formed, Lepotrema cylindricum (Wang, 1989) n. comb. (for Preptetos cylindricus) and Lepotrema navodonis (Shen, 1986) n. comb. (for Lepocreadium navodoni). With the exception of a handful of ambiguous records, the evidence is compelling that the host-specificity of species in this genus is overwhelmingly oioxenous or stenoxenous. This renders the host distribution in three orders and ten families especially difficult to explain as many seemingly suitable hosts are not infected. Multi-loci molecular data (ITS2 rDNA, 28S rDNA and cox1 mtDNA) demonstrate that Lepotrema is a good generic concept, but limited variability in sequence data and differences in phylogenies produced for different gene regions make relationships within the genus difficult to define.     

Diversity of endophytic fungal and bacterial communities in <Emphasis Type="Italic">Ilex paraguariensis</Emphasis> grown under field conditions

The composition and diversity of the endophytic community associated with yerba mate (Ilex paraguariensis) was investigated using culture-depending methods. Fungi were identified based on their micromorphological characteristics and internal transcribed spacer rDNA sequence analysis; for bacteria 16S rDNA sequence analysis was used. Fungal and bacterial diversity did not show significant differences between organ age. The highest fungal diversity was registered during fall season and the lowest in winter. Bacterial diversity was higher in stems and increased from summer to winter, in contrast with leaves, which decreased. The most frequently isolated fungus was Fusarium, followed by Colletotrichum; they were both present in all the sampling seasons and organ types assayed. Actinobacteria represented 57.5 % of all bacterial isolates. The most dominant bacterial taxa were Curtobacterium and Microbacterium. Other bacteria frequently found were Methylobacterium, Sphingomonas, Herbiconiux and Bacillus. Nitrogen fixation and phosphate solubilization activity, ACC deaminase production and antagonism against plant fungal pathogens were assayed in endophytic bacterial strains. In the case of fungi, strains of Trichoderma, Penicillium and Aspergillus were assayed for antagonism against pathogenic Fusarium sp. All microbial isolates assayed showed at least one growth promoting activity. Strains of Bacillus, Pantoea, Curtobacterium, Methylobacterium, Brevundimonas and Paenibacillus had at least two growth-promoting activities, and Bacillus, Paenibacillus and the three endophytic fungi showed high antagonistic activity against Fusarium sp. In this work we have made a wide study of the culturable endophytic community within yerba mate plants and found that several microbial isolates could be considered as potential inoculants useful for improving yerba mate production.     

New and little-known crickets of the subfamily phalangopsinae (Orthoptera,Gryllidae). 9. The african genus <Emphasis Type="Italic">Phaeophilacris</Emphasis>

The genus Phaeophilacris Walk. is divided into two subgenera: Phaeophilacris s. str. and Speluncacris Sjöst. The latter name, originally proposed as a generic one and synonymized with Phaeophilacris s. l., is restored as a subgeneric name. The former genera Opilionacris Sjöst. and Cavernacris Sjöst. previously synonymized with Phaeophilacris s. l. are considered as synonyms of the subgenus Speluncacris. Four new species (Ph. (S.) malawi sp. n., Ph. (S.) boldyrevi sp. n., Ph. (S.) dmitrievi sp. n., and Ph. (S.) gigantea sp. n.) and three new subspecies (Ph. (S.) bredoides mangochi subsp. n., Ph. (S.) bredoides mutinondo subsp. n., and Ph. (Ph.) townsendi aethiopica subsp. n.) of the genus Phaeophilacris are described.     

Two new species of <Emphasis Type="Italic">Haplorchoides</Emphasis> Chen, 1949 (Digenea: Heterophyidae) infecting an Australian siluriform fish, <Emphasis Type="Italic">Neoarius graeffei</Emphasis> Kner &amp; Steindachner

Combined morphological and molecular analyses are used to characterise two new species of Haplorchoides Chen, 1949 (Digenea: Heterophyidae) from an Australian siluriform fish. Haplorchoides maiwariensis n. sp. and H. daguilarensis n. sp. are described from the intestine of the Blue salmon catfish, Neoarius graeffei (Kner & Steindachner) (Siluriformes: Ariidae) from tidal reaches of the Brisbane River and from a freshwater creek off Lake Wivenhoe, Kipper Creek, Australia. The two new species most obviously differ from all previously described species of Haplorchoides in possessing clearly and reliably tripartite seminal vesicles. The two new species differ in the size and distribution of vitelline follicles, the size of the pigment granules and the form of the ventral sucker. Previously described species of Haplorchoides have been reported from Africa and Asia, principally from bagrid, schilbeid, silurid and sisorid catfishes; these are the first species reported from an ariid catfish. Complete ITS2 and partial 28S ribosomal DNA data were generated for both new species. The two species differ from each other by eight base pairs in the ITS2 region, and by 13 bp for the 28S region. The 28S rDNA sequence of H. daguilarensis agrees with a previously reported sequence from an unidentified species of Haplorchoides collected from N. graeffei in Lake Wivenhoe, Australia; we identify this previous report as relating to H. daguilarensis.     

Four new species of <Emphasis Type="Italic">Paradiscogaster</Emphasis> Yamaguti, 1934 (Digenea: Faustulidae) from batfishes (Perciformes: Ephippidae) on the Great Barrier Reef,Australia

Examination of three species of batfishes (Teleostei: Epphippidae) from off Lizard and Heron Islands on the Great Barrier Reef led to the discovery of specimens of the trematode genus Paradiscogaster Yamaguti, 1934 (Digenea: Faustulidae). Morphological analysis demonstrated that the new specimens represented four morphotypes which we interpret to be new species: Paradiscogaster martini n. sp., P. vichovae n. sp. and P. brayi n. sp. from Platax orbicularis (Forsskål) and P. pinnatus (Linnaeus) off Lizard Island, and P. nitschkei n. sp. from P. teira (Forsskål) off Heron Island. Published material was re-examined and the specimens identified as P. chaetodontis okinawensis Yamaguti, 1971 from P. pinnatus from Okinawa, Japan, actually represent the new species P. brayi n. sp., demonstrating that some species of Paradiscogaster have wide geographical distributions. ITS2 rDNA data for the four morphotypes differ by 4–39 base pairs confirming the delineation of the four species proposed. A feature of this study is the recognition of Platax spp. as an important host group for Paradiscogaster, with the new species placing them as the second richest host group for these parasites after the Chaetodontidae.     

A new species of <Emphasis Type="Italic">Ophiognomonia</Emphasis> from Northern China inhabiting the lesions of chestnut leaves infected with <Emphasis Type="Italic">Diaporthe eres</Emphasis>

A fungus inhabiting the lesions of chestnut leaves infected with Diaporthe eres Nitschke was identified as a new species, Ophiognomonia castaneae (Ophiognomonia, Gnomoniaceae, Diaporthales). In this study, morphological characterization and multi-gene analysis of three markers [internal transcribed spacer regions nuc rDNA ITS1-5.8S-ITS2 (ITS), guanine nucleotide-binding protein subunit beta gene (MS204), and translation elongation factor 1-α gene (tef1-α)] were used to identify this previously undescribed fungus. The teleomorphic and anamorphic stages are described and illustrated.     

Detection of pea wilt pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">pisi</Emphasis> using DNA-based markers

Identification of the fungus Fusarium oxysporum f. sp. pisi (Fop), the causal organism of wilt disease of pea, is a time consuming and arduous task. Diagnosis of Fop by traditional means requires more than 2 months and involves two steps, identification of species using morphological characters and formae specialis ‘pisi’ using pathogenicity assays. The ambiguous morphological differences between F. solani and F. oxysporum further complicate the diagnosis of F. oxysporum. A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) based method was developed to detect Fop from India. A PCR–RFLP marker, HPACAPS1₃₈₀, generated after restriction of 28S rDNA region with enzyme MvaI, detected accurately the Fop among several other fungi with detection sensitivity of 5 fg of Fop genomic DNA. In a mixture of Fop and pea DNA, the sensitivity was 500 pg of Fop DNA in 50 ng of pea DNA. The assay was further refined to detect the Fop from infected tissues and infested soil. The current assay can detect Fop from culture, plant tissues and soil in a considerably shorter period of time compared to traditional methods.     

Two monorchiid species from the freckled goatfish, <Emphasis Type="Italic">Upeneus tragula</Emphasis> Richardson (Perciformes: Mullidae), in Moreton Bay,Australia, including a proposal of a new genus

Two monorchiid species are reported from the freckled goatfish, Upeneus tragula Richardson, from Moreton Bay, Queensland, Australia. Specimens of a species new to science were most morphologically similar to species of the genus Timonia Bartoli & Prevot, 1966, but significant differences in the arrangement of the testes (symmetrical vs oblique) and morphology of the terminal organ (bipartite vs unipartite) necessitate the proposal of a new genus; Madhavia n. g. is proposed for M. fellaminutus n. sp. Specimens of the second species are identified as Parachrisomon delicatus (Manter & Pritchard, 1964) Madhavi, 2008, extending its known range from Hawaii to Australia. Complete ITS2 and partial 28S rDNA sequence data were generated for both species and analysed with those for other monorchiids available on GenBank. Phylogenetic analyses of the 28S rDNA dataset showed that both genera are distinct from other sequenced monorchiids, but overall the resolution between genera is poor and more sequence data are required to elucidate relationships within the family. We propose to transfer Timonia stunkardi (Ahmad, 1985) and Timonia vinodae (Ahmad, 1987) to the genus Neotimonia Madhavi, 2008, as Neotimonia stunkardi (Ahmad, 1985) n. comb. and Neotimonia vinodae (Ahmad, 1987) n. comb. Additionally, we were unable to locate any literature on Parachrisomon brotulidorum (Toman, 1973) Madhavi, 2008 and consider this species as nomen nudum.     

Three new species of <Emphasis Type="Italic">Plagioporus</Emphasis> Stafford, 1904 from darters (Perciformes: Percidae), with a redescription of <Emphasis Type="Italic">Plagioporus boleosomi</Emphasis> (Pearse, 1924) Peters, 1957

A form of Plagioporus Stafford, 1904 is described from the intestine of three North American species of darters (Perciformes: Percidae) from River West Twin, Wisconsin, USA, that we consider to be conspecific with Plagioporus boleosomi (Pearse, 1924) Peters, 1957 based on similarities in the sucker ratio, extent of the forebody, shape and position of the testes, vitellarium distribution and terminal genitalia. Three new species of Plagioporus are described from the intestine of darters as follows: Plagioporus fonti n. sp. from Percina nigrofasciata Agassiz in Florida, USA, Plagioporus limus n. sp. from Etheostoma squamosum Distler in Arkansas, USA and Plagioporus aliffi n. sp. from Etheostoma blennioides newmanni Miller in Arkansas, USA. Morphologically Plagioporus fonti n. sp., Plagioporus limus n. sp. and Plagioporus aliffi n. sp. are most similar to one another and to P. boleosomi, Plagioporus lepomis Dobrovolny, 1939 and ‘P. etheostomae’, a nomen nudum for a species described from Etheostoma blennioides Rafinesque in Kentucky, USA, all of which are collectively distinguished from congeners in having a combination of confluent vitellarium in the post-testicular space and absence of vitelline follicles with their entire length distributed in the forebody. Plagioporus fonti n. sp., P. limus n. sp. and P. aliffi n. sp. are respectively distinguished from one another and their closest congeners in having the anterior extent of the vitellarium in the anterior half of forebody to slightly anterior to the ventral sucker as opposed to one approximately at the level of the posterior margin of the ventral sucker, possession of an excretory vesicle reaching the anterior testis as opposed to one only reaching the posterior testis and having a longer than wide oral sucker and a wider than long ventral sucker. A Bayesian inference (BI) analysis of partial 28S rDNA sequences was conducted using the three new species and 24 sequences of opecoelids retrieved from GenBank, including ten species of Plagioporus. Plagioporus aliffi n. sp., Plagioporus fonti n. sp. and P. boleosomi comprised a moderately supported sister group to a clade containing all species of Plagioporus except Plagioporus limus n. sp. and Plagioporus shawi (Mcintosh, 1939) Margolis, 1970. Plagioporus limus and in turn P. shawi were resolved as sister to all other congeners with high and moderate support, respectively.