A new species of <Emphasis Type="Italic">Zygophiala</Emphasis> associated with the flyspeck complex on apple from China

Zygophiala qianensis is described as a new fungal species associated with the cuticle of apple fruit sampled from an orchard in Shaanxi Province, China. Conidiophores were separate, arising from superficial hyphae, erect, scattered, subcylindrical, irregularly flexuous, consisting of four parts: a hyaline supporting cell that gives rise to a smooth, dark brown stipe, terminating in a finely verruculose, medium brown apical cell that gives rise to (1–)2(–3) medium brown, finely verruculose, doliiform to ellipsoidal, polyblastic conidiogenous cells, with 1–2 prominent scars, apical and lateral, darkened, thickened. Conidia were solitary, fusiform to obclavate, hyaline, smooth and thick-walled, transversely (0–)1(–7)-septate, mostly 1–2-septate, prominently constricted at the septum; apex obtuse, base subtruncate, with a darkened, thickened hilum. Zygophiala qianensis is compared morphologically to other species of Zygophiala, and a phylogenetic analysis of their DNA sequence data is presented.     

Four species of Zygophiala (Schizothyriaceae, Capnodiales) are associated with the sooty blotch and flyspeck complex on apple

Sooty blotch and flyspeck (SBFS) is a complex of fungi that cause late-season blemishes of apple and pear fruit that cosmetically damage the cuticle, which result in fruit that are unacceptable to consumers. Previous studies reported that a single, wide-host-range species, Schizothyrium pomi (presumed anamorph Zygophiala jamaicensis), caused flyspeck on apple. In the present study we compared morphology and DNA phylogeny (ITS, LSU) of 139 fungal strains isolated from flyspeck signs from 39 apple orchards in 14 midwestern and eastern states (USA). Parsimony analysis, supported by cultural characteristics and morphology in vitro, provided support to delimit the flyspeck isolates into four species of Zygophiala, two of which are known to be sexual. Three of these species are described as new. Based on DNA phylogeny, species of Schizothyrium were shown to cluster with members of the genus Mycosphaerella in the Capnodiales, having similar asci and ascospores but morphologically distinct ascomata. These data question the value of ascomatal morphology at the ordinal level, although it still appears to be relevant at the family level, delimiting the thyrothecial Schizothyriaceae from other families in the Capnodiales.     

Molecular and Morphological Analysis Reveals Five New Species of Zygophiala Associated with Flyspeck Signs on Plant Hosts from China

Species in the genus Zygophiala are associated with sooty blotch and flyspeck disease on a wide range of hosts. In this study, 63 Zygophiala isolates collected from flyspeck colonies on a range of plants from several regions of China were used for phylogeny, host range and geographic distribution analysis. Phylogenetic trees were constructed on four genes - internal transcribed spacer (ITS), partial translation elongation factor 1-alpha (TEF), β-tubulin (TUB2), and actin (ACT) – both individually and in combination. Isolates were grouped into 11 clades among which five new species, Z. emperorae, Z. trispora, Z. musae, Z. inaequalis and Z. longispora, were described. Species of Zygophiala differed in observed host range and geographic distribution. Z. wisconsinensis and Z. emperorae were the most prevalent throughout the sampled regions of China, whereas Z. trispora, Z. musae, Z. inaequalis and Z. longispora were collected only in southern China. The hosts of Z. wisconsinensis and Z. emperorae were mainly in the family Rosaceae whereas Z. trispora, Z. musae, Z. inaequalis and Z. longispora were found mainly on banana (Musa spp.). Cross inoculation tests provided evidence of host specificity among SBFS species.     

Redescription of the East Asian bagrid catfish, <Emphasis Type="Italic">Pseudobagrus kyphus</Emphasis> Mai, 1978, and a new record from China

 A small species (<90 mm SL) of the bagrid catfish, Pseudobagrus kyphus Mai, 1978, originally described from northern Vietnam, is newly recorded from China and redescribed in detail. The species is distributed in rivers flowing into Beibu Gulf (Gulf of Tonkin); i.e., the Song Hong River basin in northern Vietnam, and rivers on the southern side of Mts. Shiwan–Dashan in southernmost Guangxi Zhuangzu Autonomous Region, China. Partial nucleotide sequences of the mitochondrial DNA are also described and compared among the populations.     

Characterization of <Emphasis Type="Italic">Penicillium</Emphasis> Species by Ribosomal DNA Sequencing and BOX,ERIC and REP-PCR Analysis

The genus Penicillium is one of the largest and widely distributed fungal genera described to date. As a result, its taxonomic classification and species discrimination within this genus has become complicated. In this study, 52 isolates that belonged to the Penicillum genus and other related genera were characterized using two DNA-based methods: (i) analysis of the nucleotide sequences of internal transcribed spacers in ribosomal DNA and (ii) analysis of DNA fingerprints that were generated by polymerase chain reactions with specific primers for enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP) sequences, and BOX elements. Using both methods, Penicillium species were discriminated from other fungal genera. Furthermore, Penicillium species that include strains which are used as biocontrol agents, such as P. glabrum, P. purpurogenum, and P. oxalicum, could be distinguished from other Penicillium species using these techniques. Based on our findings, we propose that a polyphasic approach that includes analysis of the nucleotide sequences of ribosomal DNA and detecting the presence of highly conserved, repeated nucleotide sequences can be used to determine the genetic relationships between different Penicillium species. Furthermore, we propose that our results can be used as a start point to develop a strategy to monitor the environmental presence of particular strains of Penicillium species when they are used as biocontrol agents.     

A new species of Scolecobasidium associated with the sooty blotch and flyspeck complex on banana from China

Sooty blotch and flyspeck is caused by a complex of epiphytic fungi on diverse hosts. A fungus morphologically similar to Scolecobasidium humicola was isolated from a banana fruit exhibiting SBFS signs in Hainan, China. ITS and LSU sequences of rDNA corroborated that it was a Scolecobasidium species. The results of LSU rDNA analysis support the interpretation that Ochroconis is synonymous with Scolecobasidium. Five new combinations, S. anomalum, S. calidifluminale, S. gallopavum, S. gamsii, and S. lascauxense, were proposed. A new species, S. musae, was described based on morphological characteristics and sequence comparison with previous species.     

First Record of Didymella anemones sp. nov. Teleomorph of Ascochyta anemones,the Causal Agent of Leaf Spots on Pulsatilla korreana in China

The teleomorph of Ascochyta anemones has been recorded for the first time on overwintering windflower stalk in Liaoning, China. The features of this new species were described, and it was distinguished from other species to which it is similar. The morphological characteristics of this species, such as asci and ascospores, phylogenetic analysis based on internal transcribed spacer sequences, comparison with similar fungi so far described from Ranunculaceae indicated that the teleomorph is an undescribed species of Didymella.     

A New Species of <Emphasis Type="Italic">Rhytisma</Emphasis> Causes Tar Spot on <Emphasis Type="Italic">Comarostaphylis arbutoides</Emphasis> (Ericaceae) in Panama

A fungus causing tar spots on leaves of Comarostaphylis arbutoides (Ericaceae) in Panama is described as a new species, Rhytisma panamense. The fungus forms gregarious black stromata on pale yellow spots on the adaxial side of leaves. Its ascomata develop from unilocular or multilocular stromata. An analysis of a combined dataset of DNA sequences from LSU to ITS rDNA supports the placement of the species in the genus Rhytisma.     

Expansion of the sooty blotch and flyspeck complex on apples based on analysis of ribosomal DNA gene sequences and morphology

Sooty blotch and flyspeck (SBFS) is a late-season disease of apple and pear fruit that cosmetically damages the cuticle, resulting in produce that is unacceptable to consumers. Previous studies reported that four species of fungi comprise the SBFS complex. We examined fungal morphology and the internal transcriber spacer (ITS) and large subunit (LSU) regions of rDNA of 422 fungal isolates within the SBFS complex from nine orchards in four Midwestern states (USA) and compared them to previously identified species. We used LSU sequences to phylogenetically place the isolates at the order or genus level and then used ITS sequences to identify lineages that could be species. We used mycelial and conidial morphology on apple and in culture to delimit putative species. Thirty putative species found among the Midwest samples were shown to cause SBFS lesions on apple fruit in inoculation field trials. Among them Peltaster fructicola and Zygophiala jamaicensis have been associated previously with SBFS in North Carolina. The LSU analyses inferred that all 30 SBFS fungi from Midwestern orchards were Dothideomycetes; one putative species was within the Pleosporales, 27 were within Dothideales, and two putative species could not be placed at the ordinal level. The LSU sequences of 17 Dothideales species clustered with LSU sequences of known species of Mycosphaerella.     

Two new species of <Emphasis Type="Italic">Trichoderma</Emphasis> from Yunnan,China

Two new species of the fungal genus Trichoderma, Trichoderma compactum and Trichoderma yunnanense, isolated from rhizosphere of tobacco in Yunnan Province, China are described based on morphological characters and phylogenetic analyses of nucleotide sequences. Our DNA sequences included the internal transcribed spacer (ITS) regions of the rDNA cluster (ITS1 and ITS2), and partial sequences of the translation elongation factor 1-alpha (tef1) and a fragment of the gene coding for endochitinase 42 (ech42). The analyses show that T. compactum belongs to the Harzianum clade, and T. yunnanense belongs to the Hamatum clade.     

Discovery of <Emphasis Type="Italic">Ophiostoma tsotsi</Emphasis> on <Emphasis Type="Italic">Eucalyptus</Emphasis> wood chips in China

Ophiostoma species such as O. quercus are the most frequent causal agents of sapstain of freshly felled hardwood timber and pulpwood. Many species are regarded as economically important agents of wood degradation. The aim of this study was to identify a collection of Ophiostoma isolates, resembling O. quercus, found on stained Eucalyptus pulpwood chips in China. DNA sequences of the internal transcribed spacer regions, including the 5.8S region, of the ribosomal DNA, and parts of the β-tubulin and elongation factor-1α genes, revealed that the isolates were not O. quercus. Surprisingly, they represented O. tsotsi, a wound-infesting fungus recently described from hardwoods in Africa. In addition, sequence data from an isolate from agarwood in Vietnam, identified in a previous study as belonging to an unknown Pesotum species, were also shown to represent O. tsotsi. A high level of genetic variability was observed among isolates of both O. quercus and O. tsotsi. This was unexpected and suggests that both species have been present in Asia for a significant amount of time.     

Molecular and morphological identification of truffle-producing <Emphasis Type="Italic">Tuber</Emphasis> species in New Zealand

Molecular and morphological techniques were used to examine New Zealand ascomycetous truffle (Tuber spp.) samples deposited in the Plant & Food Research and Landcare Research Fungi Herbarium collections. Truffles have been found on the roots of many Northern Hemisphere tree species growing in New Zealand, but not on indigenous plant species. Comparisons of ribosomal DNA sequences proved to be a simple and rapid method to identify the Tuber species. Tuber maculatum was by far the predominant species in New Zealand, and was distributed throughout the country. A single truffle sample from Christchurch was identified as T. rufum. Two other groups of truffle samples from Pinus spp. were closely related to anonymous Northern Hemisphere Tuber sequences. Ascocarps with these sequences have not previously been described. Specific primers for the PCR detection of these Pinus isolates were developed. None of these Tuber species accidentally introduced to New Zealand is of economic value.     

A new species of Devriesia causing sooty blotch and flyspeck on Rubber Trees in China

Sooty blotch and flyspeck is a fungal disease complex that colonizes a wide range of plants. In a survey of host plants for sooty blotch and flyspeck fungi, we obtained an isolate associated with discrete flyspeck symptom on stems of the Rubber Trees, Ficus elastica, from Haikou, Hainan, China. A pure culture was obtained, and the nuclear ITS region of rDNA was amplified by PCR. Molecular phylogeny inferred from ITS sequence data suggested that it was close to Devriesia strelitziae, reported from South Africa. Morphologically, it was characterized by longer conidia, more conidial septa, and having chlamydospores. Therefore, we describe our isolate as a novel species of Devriesia.     

Two new species of Antispila Hübner (Lepidoptera: Adeloidea: Heliozelidae) associated with Hydrangea (Hydrangeaceae) from China

The genus Antispila Hübner, [1825] is poorly recorded from China. Two new Antispila species, A. longcangensis, sp. nov. and A. emeishanensis, sp. nov. feeding on Hydrangeaceae, are described from Sichuan, China. The adult male and female, genitalia and venation of both species are illustrated. Diagnostic characters of these species are compared with other described Antispila species from China. DNA barcodes sequences of the two new species are also provided.     

DNA barcoding and morphology reveal exceptional species diversity of Scoparia (Lepidoptera: Crambidae) from the Hailuogou Glacier area,China

The genus Scoparia from the Hailuogou Glacier area in the south‐eastern fringe of the Tibetan Plateau of south‐western China is studied, nine species are revealed by combining DNA barcoding (658 bp of the mitochondrial gene cytochrome c oxidase subunit I, COI) and morphology. Studies show that Scoparia species from the Hailuogou Glacier area are 37.5% as many as all the previously known congeners in China. Six species are described as new to science: S coparia simplicissima Li sp. nov. , S coparia tribulosa Li sp. nov. , S coparia longispina Li sp. nov. , S coparia gibbosa Li sp. nov. , S coparia globosa Li sp. nov. , and S coparia annulata Li sp. nov. The female of Scoparia metaleucalis is described for the first time herein. All species are either diagnosed or described and illustrated, and a map of China showing the topography and localities where Scoparia species are recorded is also provided, including the species number of all the recorded localities. The results reveal the exceptional species diversity of Scoparia from the Hailuogou Glacier area, as well as demonstrate that the integration of DNA barcoding and morphological approaches is highly effective for indentifying scopariine moths in the Hailougou glacier area. All the studied specimens are deposited in the Insect Museum, Jiangxi Agricultural University, Nanchang, China. © 2014 The Linnean Society of London     

Out of South America: phylogeny of non‐biting midges in the genus Labrundinia suggests multiple dispersal events to Central and North America

Non‐biting midges of the genus Labrundinia (Chironomidae: Tanypodinae) are minute dipterans with immature stages living in a variety of unpolluted water bodies, from small streams and ponds to lakes and bays. Extensively recorded in ecological studies, the genus comprises 39 species, all except one described from areas outside the Palearctic region. Internal structure among Labrundinia species was postulated by S. S. Roback, who recognized four species groups based on morphological characters of immature stages. We examined phylogenetic relationships among known Labrundinia species using partial DNA sequences of the nuclear protein‐coding gene CAD and morphological characters. Both analyses with Bayesian inference and parsimony methods recovered the monophyly of Labrundinia, strongly supported by five morphological synapomorphies. Internal relationships within the genus partly supported Roback's species groups with the addition of later described species. Biogeographical inferences were obtained by applying Bayesian binary MCMC (BBM) analysis and favoured a scenario where Labrundinia had its initial diversification in the Neotropical region and that current presence in the Nearctic region and southern South America is due to subsequent dispersal.     

Two species of Lirula on Abies from Yunnan, Southwest China

Based on a recent investigation of species of the Rhytismatales in the eastern Himalayas in China, two species of Lirula on Abies were found, one of which is described as a new species, Lirula yunnanensis, which differs from Lirula japonica mainly by ascomatal structure and conidiomatal features, while the other is Lirula exigua, a new record for China. An analysis of the combined sequences of ITS + LSU r DNA showed that Lirula macrospora and Lophodermium piceae were closely related and that Lirula yunnanensis and Lophodermium autumnale were grouped together. The genus of Lirula should be redefined based on more molecular data. Lirula exigua was distantly related to Lirula species and its systematic position has to be confirmed by further molecular data. A key for known species of Lirula worldwide is provided.     

Characterization of an <Emphasis Type="Italic">Eco</Emphasis>RI family of satellite DNA from two species

We have cloned and sequenced a 321bp band of repetitive DNA from Eptesicus fuscus and E. serotinus observed after gel electrophoresis of EcoRI digested genomic DNA in both species. Southern blot analysis of genomic DNA (from both species) digested with the same enzyme showed the existence of a ladder pattern indicating that the repetitive DNA is arrayed in tandem. The repetitive sequences have a monomer unit of 321bp which is composed of two subunits of 160bp, suggested by the existence of a 160bp band in the ladder of E. fuscus and by the presence of some direct repeats found in the analysis of the consensus sequence. Analysis of the methylation status demonstrated that cytosines in CCGG sequences in this satellite DNA are methylated in E. fuscus but not in the E. serotinus. Alignment of the sequenced clones showed that several nucleotide positions are diagnostic species-specific and consequently the phylogenetic analysis grouped the monomer units from both species in two clearly separated groups.     

Early larvae of <Emphasis Type="Italic">Zesticelus profundorum</Emphasis> (family Cottidae) identified using DNA barcoding

Early larvae of Flabby Sculpin, Zesticelus profundorum, are described using DNA barcoding methods to identify specimens collected in the Gulf of Alaska and Bering Sea. Based on the low myomere count, distribution of larvae, and vertebral counts in adults, we selected Zesticelus profundorum and Sebastolobus macrochir as good potential candidates for genetic comparison. We used DNA samples from voucher specimens of each candidate taxon to identify species-specific sequences. An exact match in sequence data allowed us to positively identify these larvae as Zesticelus profundorum, a rare deepwater species of the family Cottidae.     

<Emphasis Type="Italic">Gallus gallus</Emphasis> aggrecan gene-based phylogenetic analysis of selected avian taxonomic groups

Mitochondrial DNA (mtDNA) sequences remain the most widely used for phylogenetic analysis in birds. A major limitation of mtDNA sequences, however, is that mitochondria genes are inherited as a single linkage group. Here we describe the use of a 540-bp DNA sequence corresponding to the G3 domain of Gallus gallus nuclear aggrecan gene (AGC1) for phylogenetic analysis of the main groups of Galliformes including Phasianidae, Numididae, and Odontophoridae. We also included species from Cracidae and Megapodiidae which are considered by some as Craciformes and others, including here as Galliformes. The uncorrected sequence divergence of the G3 fragments ranges from 1 among the grouses to 36% between some of the distant groups within Galliformes. These sequences contain 39–48% AT nucleotides and the ratios of transition versus transversion are above 1.5 in majority of the comparisons. Using G3 sequences from an Anseriform, Oxyura jamaicensis, as out-groups, phylogenetic trees were obtained using maximum parsimony and distance algorithms and bootstrap analyses. These trees were consistent with those described using Avian sarcoma and leucosis virus gag genes and those from amino acid sequences of hemoglobin and lysozyme c. Our data also support relationships among Galliformes which were defined using mtDNA sequences. In addition to the general support of the five main families of Galliformes, our data are also consistent with previous work that showed Francolinus africanus and Gallus gallus are in the same clade and that Tetraoninae is a well-supported monophyletic subfamily within Phasianidae. The results presented here suggest that the AGC1 sequences meet the criterion of novel nuclear DNA sequences that can be used to help resolve the relationships among Galliformes.