<Emphasis Type="Italic">Phragmidium</Emphasis> species parasitizing species of Rosaceae in Tibet,China, with descriptions of three new species

Phragmidum spp. are rust fungi that are serious pathogens of plants in the Rosaceae. We characterized 15 Phragmidium species from Tibet, including 12 previously described and 3 new species. All the taxa, including the three new species (Ph. chayuensis, Ph. cibanum, Ph. zangdongii) are described and illustrated based on morphological characteristics. These taxa have multi-celled teliospores with firm pedicels. Phragmidium chayuensis is characterized by (5)7–9-celled teliospores with short papillae. Phragmidium cibanum is characterized by smooth teliospores that are distinctly constricted at the septa. Phragmidium zangdongii is characterized by large uredinia and 11–13-celled teliospores. Five types of urediniospore-surface structures were identified based on the gross shape of ornamentations and their distribution on the urediniospore wall. Molecular sequence data from the LSU rDNA analysis showed that the new taxa formed distinct clades independent from previously recorded species represented by LSU rDNA sequence data. The phylogenetic tree indicated that Phragmidium species are highly host specific.     

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.     

Taxonomy and phylogenetic position of <Emphasis Type="Italic">Phyllactinia takamatsui</Emphasis>, a newly described powdery mildew on cotoneaster,based on molecular and morphological data

The newly recognised powdery mildew species Phyllactinia takamatsui on Cotoneaster nummularius (Rosaceae) is described and illustrated. This species, collected in Kerman Province, Iran, is well characterised by its conidial morphology and rDNA ITS sequences clearly different from allied species. Conidia are broadly ellipsoid to subcylindrical, i.e. they are not clavate-spathuliform as in most Phyllactinia species. The rDNA ITS sequence analysis showed that this species is closely allied to other species described on hosts belonging to Rosaceae, such as Ph. mali and Ph. pyri-serotinae. The ITS sequence of P. takamatsui was 92 to 94 % similar to that of the closest known relatives. The new species is described in detail, illustrated and compared with other similar taxa.     

Additions to <Emphasis Type="Italic">Lindgomyces</Emphasis> (Lindgomycetaceae,Pleosporales, Dothideomycetes), including two new species occurring on submerged wood from North Carolina,USA, with notes on secondary metabolite profiles

Two new species of freshwater ascomycetes belonging to the genus Lindgomyces (Pleosporales, Dothideomycetes) are described and illustrated from submerged wood in North Carolina, USA. Lindgomyces carolinensis is characterized by immersed to erumpent ascomata, fissitunicate broadly cylindrical to clavate asci, and fusiform ascospores with acute ends surrounded by a large, fusiform gelatinous sheath. Lindgomyces cigarospora morphologically differs from L. carolinensis in that its ascospores are fusiform to cylindrical with rounded ends, without a large fusiform gelatinous sheath. These two new species nest in the family Lindgomycetaceae based on analyses of combined SSU and LSU rDNA sequence data. Phylogenetic analyses using ITS sequence data support the establishment of the new taxa as separate species within Lindgomyces. In addition to the new species, we report new ITS sequence data for L. cinctosporus and L. griseosporus from France, and L. ingoldianus from North Carolina, USA. We report a video exhibiting fissitunicate ascus dehiscence in L. carolinensis showing ascospore discharge and unraveling of the gelatinous sheath in real time. Chemical analysis of the organic extracts of L. carolinensis and L. cigarospora resulted in two known cyclodepsipeptides, Sch 378161 and Sch 217048. The in situ spatial mapping of these secondary metabolites on fungal cultures indicates the presence of both compounds on the surface of mycelia, as well as being exuded into the surrounding agar.     

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.     

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.     

<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.     

Phylogenetic analysis of <Emphasis Type="Italic">Plectosphaerella</Emphasis> species based on multi-locus DNA sequences and description of <Emphasis Type="Italic">P. sinensis</Emphasis> sp. nov.

Species in Plectosphaerella are well known as pathogens of several plant species causing fruit, root and collar rot and collapse. In an investigation of endophytic fungi associated with cucurbit plants in China, we isolated 77 strains belonging to the genus Plectosphaerella. To identify the isolated strains, we collected the type or reference strains of all currently accepted species in Plectosphaerella except P. oratosquillae and conducted a phylogenetic analysis. Phylogenetic analysis of the partial 28S rDNA sequences showed that all species in Plectosphaerella were located in one clade of Plectosphaerellaceae. Based on multi-locus phylogenetic analysis of the ITS, CaM, EF1, TUB and morphological characteristics, all species in Plectosphaerella were well separated. Three endophytic strains from stems of Cucurbita moschata, Citrullus lanatus and Cucumis melo from North China were assigned to a new species described as P. sinensis in this paper. The new species differs morphologically from other Plectosphaerella species by irregular chlamydospores, and the dimensions of phialides and conidia. The other endophytic strains from several cucurbit plants were identified as P. cucumerina.     

Five new species of <Emphasis Type="Italic">Conocybe</Emphasis> (Agaricomycetes,Bolbitiaceae) from Russia

Five new Conocybe species are described from Russia based on collections made in different parts of the country. One species (C. olivaceopileata) belongs to sect. Conocybe, two species (C. praticola and C. coniferarum) to sect. Mixtae. The ornamented basidiospores of C. confundens and C. incerta is characteristic for sect. Ochromarasmius. All new species are described based on morphology and, in some cases, on DNA sequencing (ITS rDNA). Photos of basidiocarps, illustrations of microstructures and a comparison with similar taxa are given.     

Looking beneath the tip of the iceberg: diversification of the genus <Emphasis Type="Italic">Epimeria</Emphasis> on the Antarctic shelf (Crustacea,Amphipoda)

The amphipod genus Epimeria is very speciose in Antarctic waters. Although their brooding biology, massive and heavily calcified body predict low dispersal capabilities, many Epimeria species are documented to have circum-Antarctic distributions. However, these distribution records are inevitably dependent on the morphological species definition. Yet, recent DNA evidence suggests that some of these Epimeria species may be complexes of species with restricted distributions. Mitochondrial COI and nuclear 28S rDNA sequence data were used to infer evolutionary relationships among 16 nominal Epimeria species from the Antarctic Peninsula, the eastern Weddell Sea and the Adélie Coast. Based on this phylogenetic framework, we used morphology and the DNA-based methods GMYC, bPTP and BPP to investigate species boundaries, in order to revise the diversity and distribution patterns within the genus. Most of the studied species appeared to be complexes of pseudocryptic species, presenting small and previously overlooked morphological differences. Altogether, 25 lineages were identified as putative new species, increasing twofold the actual number of Antarctic Epimeria species. Whereas most of the species may be geographically restricted to one of the three studied regions, some still have very wide distribution ranges, hence suggesting a potential for large-scale dispersal.     

Identification of <Emphasis Type="Italic">Bacillus</Emphasis> Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, <Emphasis Type="Italic">recA</Emphasis>, <Emphasis Type="Italic">rpoB</Emphasis> Gene Sequencing and RAPD-PCR

Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products.     

Four new species of <Emphasis Type="Italic">Tubeufia</Emphasis> (Tubeufiaceae,Tubeufiales) from Thailand

Tubeufia was named for a sexual morph taxon. However, several asexual morph species have been accommodated in this genus as well. In our study, four new species of Tubeufia, viz. T. filiformis, T. latispora, T. laxispora and T. mackenziei, are described and illustrated. The phylogenetic placement of the new species is confirmed by analysis of combined ITS, LSU and TEF1α sequence data. A key to Tubeufia species is provided and the new species are compared with similar fungal taxa.     

Multi-gene phylogenetic analyses reveals <Emphasis Type="Italic">Neohelicosporium</Emphasis> gen. nov. and five new species of helicosporous hyphomycetes from aquatic habitats

Helicosporous hyphomycetes are a morphologically allied group of Tubeufiales. We introduce a new helicosporous genus, Neohelicosporium, with five new species, Neohelicosporium aquaticum, N. guangxiense, N. hyalosporum, N. parvisporum, and N. thailandicum, based on morphological and phylogenetic evidence. The RPB2 protein gene data are provided to analyze their phylogeny in Tubeufiales. Phylogenetic analyses of combined ITS, LSU, RPB2, and TEF1α sequence data from 13 new isolates of Neohelicosporium provided evidence to support the establishment of the new taxa. The morphological characters of Neohelicosporium that differentiate it from other helicosporous species are compared and discussed.     

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.     

<Emphasis Type="Italic">Calcarisporium xylariicola</Emphasis> sp. nov. and introduction of <Emphasis Type="Italic">Calcarisporiaceae</Emphasis> fam. nov. in Hypocreales

During a survey of fungicolous fungi, a novel taxon from the surface of stroma of an unidentified Xylaria species was collected. Phylogenetic analyses showed that this taxon clustered with Calcarisporium sp. and C. arbuscula isolates, but was resolved as a distinct species. A detailed morphological examination coupled with phylogenetic analysis indicated that the taxon represented a new species. Calcarisporium xylariicola sp. nov. is thus introduced. The new taxon is characterized by short conidiophores with swollen bases and less length/width ratio of conidia that distinguish it from other Calcarisporium species. Calcarisporium is presently placed in Hypocreales genera, incertae sedis genus. Species in the genus are largely fungicolous, or occasionally caulicolous or foliicolous, and have hyaline, erect, verticillate conidiophores and sympodial, polyblastic conidiation. A phylogenetic analysis of combined SSU, ITS, LSU, TEF and RPB2 sequence data from Calcarisporium species and other taxa in Hypocreales indicate that Calcarisporium is a distinct lineage from other families. Therefore, a new family, Calcarisporiaceae, in Hypocreales is introduced.     

Two new species of <Emphasis Type="Italic">Bacciger</Emphasis> Nicoll, 1914 (Trematoda: Faustulidae) in species of <Emphasis Type="Italic">Herklotsichthys</Emphasis> Whitley (Clupeidae) from Queensland waters

Two new species of Bacciger Nicoll, 1914 (Faustulidae) are described infecting clupeids collected from the waters off Queensland, Australia; Bacciger minor n. sp. is described from Herklotsichthys castelnaui (Ogilby) in Moreton Bay, southern Queensland and Bacciger major n. sp. is described from Herklotsichthys quadrimaculatus (Rüppell) collected off Lizard Island, on the northern Great Barrier Reef. The two species both differ from previously described species of Bacciger in the combination of their generally elongate bodies, an entire rather than deeply lobed ovary, vitelline follicles that reach to at least the intestinal bifurcation, instead of restricted to further posteriorly but principally distributed in the hindbody, and intestinal caeca extending posteriorly well past the ventral sucker. The two new species have non-overlapping size ranges and differ in their sucker ratios, the distribution of the vitelline follicles and in the shape of the cirrus-sac. ITS2 and 28S rDNA sequence data distinguish the two new species unambiguously. Phylogenetic analysis of available 28S data show they are most closely related to Pseudobacciger cheneyae Sun, Bray, Yong, Cutmore & Cribb, 2014, also recorded off Lizard Island. These are the first faustulids reported from species of Herklotsichthys Whitley, but overall members of the Clupeidae undoubtedly harbours the richest faustulid fauna of any fish family. Baccigeroides ovatus (Price, 1934) n. comb. is proposed for Bacciger ovatus (Price, 1934) Bray & Gibson, 1980 (syn. B. opisthonema Nahhas & Cable, 1964) based on the position of the genital pore being far anteriorly removed from the ventral sucker.