Seven New Species of Eurotiales (Ascomycota) Isolated from Tidal Flat Sediments in China.

Tidal flats have been reported to contain many microorganisms and play a critical role in maintaining biodiversity. In surveys of filamentous fungi from tidal flat sediments in China, seven new species of Eurotiales were discovered and described. Morphological characteristics and DNA sequence analyses of combined datasets of the BenA, CaM, and RPB2 regions support their placements and recognition as new species. Aspergillus liaoningensis sp. nov. and A. plumeriae sp. nov. belong to sections Candidi and Flavipedes of subgenus Circumdati, and A. subinflatus sp. nov. is a member of section Cremei of subgenus Cremei. Penicillium danzhouense sp. nov., P. tenue sp. nov., and P. zhanjiangense sp. nov. are attributed to sections Exilicaulis and Lanata-Divaricata of subgenus Aspergilloides. Talaromyces virens sp. nov. is in section Talaromyces. Detailed descriptions and illustrations of these novel taxa are provided. Their differences from close relatives were compared and discussed.

New Species of Neocosmospora (Ascomycota) from China as Evidenced by Morphological and Molecular Data.

Species of Neocosmospora are commonly found in soil, plant debris, and living woody or herbaceous substrates and occasionally found in water and air. Some species are reported as saprobes, endophytes, opportunistic pathogens of plants and animals, or producers of bioactive natural products, cytotoxic compounds, and industrial enzymes. To reveal the species diversity of Neocosmospora, specimens from different provinces of China were investigated. Five new species, Neocosmospora anhuiensis, N. aurantia, N. dimorpha, N. galbana, and N. maoershanica, were introduced based on morphological characteristics and DNA sequence analyses of combined calmodulin (CAM), the internal transcribed spacer (ITS), the second largest subunit of RNA polymerase II (RPB2), and the translation elongation factor 1-α (TEF1) regions. Differences between these new species and their close relatives are compared in detail.

Three New Species of Fusicolla (Hypocreales) from China.

To explore the species diversity of the genus Fusicolla, specimens from Henan, Hubei and Jiangsu Provinces in China are examined, and three undescribed taxa are encountered. The morphological characteristics and DNA sequence analyses of the combined acl1, ITS, LSU, rpb2 and tub2 regions support their placement in Fusicolla and their recognition as new species. Fusicolla aeria sp. nov. is distinguished by the formation of abundant aerial mycelia on PDA, falcate, (1-)3-septate macroconidia 16-35 × 1.5-2.8 µm and subcylindrical, aseptate microconidia 7.5-13 × 0.8-1.1 µm. Fusicolla coralloidea sp. nov. has a coralloid colony on PDA, falcate, 2-5-septate macroconidia 38-70 × 2-4.5 µm and rod-shaped to ellipisoidal, aseptate microconidia 2-7 × 1-1.9 µm. Fusicolla filiformis sp. nov. is characterized by filiform, 2-6-septate macroconidia 28-58 × 1.5-2.3 µm and lacking microconidia. Morphological differences between these novel species and their close relatives are compared in detail. The previously recorded species of the genus in China are listed and a key to these taxa is provided.

Three New Species of Clonostachys (Hypocreales, Ascomycota) from China.

Three new species of Clonostachys are introduced based on specimens collected from China. Clonostachys chongqingensis sp. nov. is distinguished by pale yellow to pale orange-yellow perithecia with a very low papilla, clavate to subcylindrical asci possessing ellipsoidal to elongate-ellipsoidal spinulose ascospores 13-16 × 4.5-5.5 µm; it has acremonium- to verticillium-like conidiophores and ellipsoidal to rod-shaped conidia. Clonostachys leptoderma sp. nov. has pinkish-white subglobose to globose perithecia on a well-developed stroma and with a thin perithecial wall, clavate to subcylindrical asci with ellipsoidal to elongate-ellipsoidal spinulose ascospores 7.5-11 × 2.5-3.5 µm; it produces verticillium-like conidiophores and ellipsoidal to subellipsoidal conidia. Clonostachys oligospora sp. nov. features solitary to gregarious perithecia with a papilla, clavate asci containing 6-8 smooth-walled ascospores 9-17 × 3-5.5 µm; it forms verticillium-like conidiophores and sparse, subfusiform conidia. The morphological characteristics and phylogenetic analyses of combined nuclear ribosomal DNA ITS1-5.8S-ITS2 and beta-tubulin sequences support their placement in Clonostachys and their classification as new to science. Distinctions between the novel taxa and their close relatives are compared herein.

New Species of Nectriaceae (Hypocreales) from China.

Species of Nectriaceae commonly occur on living and decaying woody substrates, soil, fruitbodies of other fungi, and insects. Some of them are reported as endophytes, opportunistic pathogens of crops and humans, or producers of mycotoxins. To explore the species diversity of the family, specimens from different regions of China were collected and examined. Four novel taxa of Penicillifer, Pseudocosmospora, and Thelonectria were introduced on the basis of morphological characteristics and DNA sequence analyses of combined datasets of the act, ITS, LSU, rpb1, rpb2, tef1, and tub2 regions. Differences between the new species and their close relatives were compared and discussed.

Comparative molecular evolution of chitinases in ascomycota with emphasis on mycoparasitism lifestyle.

Chitinases are involved in multiple aspects of fungal life cycle, such as cell wall remodelling, chitin degradation and mycoparasitism lifestyle. To improve our knowledge of the chitinase molecular evolution of Ascomycota, the gene family of 72 representatives of this phylum was identified and subjected to phylogenetic, evolution trajectory and selective pressure analyses. Phylogenetic analysis showed that the chitinase gene family size and enzyme types varied significantly, along with species evolution, especially for groups B and C. In addition, two new subgroups, C3 and C4, are recognized in group C chitinases. Random birth and death testing indicated that gene expansion and contraction occurred in most of the taxa, particularly for species in the order Hypocreales (class Sordariomycetes). From an enzyme function point of view, we speculate that group A chitinases are mainly involved in species growth and development, while the expansion of genes in group B chitinases is related to fungal mycoparasitic and entomopathogenic abilities, and, to a certain extent, the expansion of genes in group C chitinases seems to be correlated with the host range broadening of some plant-pathogenic fungi in Sordariomycetes. Further selection pressure testing revealed that chitinases and the related amino acid sites were under positive selection in the evolutionary history, especially at the nodes sharing common ancestors and the terminal branches of Hypocreales. These results give a reasonable explanation for the size and function differences of chitinase genes among ascomycetes, and provide a scientific basis for understanding the evolutionary trajectories of chitinases, particularly that towards a mycoparasitic lifestyle.

Proharziane and Harziane Derivatives from the Marine Algicolous Fungus Trichoderma asperelloides RR-dl-6-11.

One new proharziane and three new harziane derivatives (1-4) together with six known ones (5-10) were isolated from the marine-alga-derived ascomycete Trichoderma asperelloides RR-dl-6-11. Their structures and relative configurations were determined via spectroscopic techniques, and the absolute configurations were ascertained by analysis of ECD curves. This is the first report on the secondary metabolites of T. asperelloides, and the new isolates (1-4), especially seco-harziane 4, greatly add to the structural diversity of harziane diterpenes as well as their precursors and catabolites. Compounds 1-5 inhibited four marine phytoplankton species, and the structure-activity relationship of harziane derivatives is analyzed.

Ascomycetes from the Qilian Mountains, China - Hypocreales.

To investigate fungi from the Qilian Mountains in Gansu Province, ascomycetous specimens were collected and hypocrealean fungi were examined. Eighteen species belonging to six genera in the families Hypocreaceae and Nectriaceae were identified, including 11 species of Hypomyces and Trichoderma in Hypocreaceae and seven species of Nectria, Stylonectria, Thelonectria, and Thyronectria in Nectriaceae. Among them, Stylonectria qilianshanensis and Trichoderma gansuanum are new to science. DNA sequence analyses of combined ACL1, ITS, RPB2, and TEF1 regions confirmed their taxonomic placements. Morphological distinctions between the new species and their close relatives are discussed. Hypomyces tremellicola is reported for the first time in China.

Two New Species and a New Chinese Record of Hypocreaceae as Evidenced by Morphological and Molecular Data.

To explore species diversity of Hypocreaceae, collections from Guangdong, Hubei, and Tibet of China were examined and two new species and a new Chinese record were discovered. Morphological characteristics and DNA sequence analyses of the ITS, LSU, EF-1α, and RPB2 regions support their placements in Hypocreaceae and the establishments of the new species. Hypomyces hubeiensis sp. nov. is characterized by occurrence on fruitbody of Agaricus sp., concentric rings formed on MEA medium, verticillium-like conidiophores, subulate phialides, rod-shaped to narrowly ellipsoidal conidia, and absence of chlamydospores. Trichoderma subiculoides sp. nov. is distinguished by effuse to confluent rudimentary stromata lacking of a well-developed flank and not changing color in KOH, subcylindrical asci containing eight ascospores that disarticulate into 16 dimorphic part-ascospores, verticillium-like conidiophores, subcylindrical phialides, and subellipsoidal to rod-shaped conidia. Morphological distinctions between the new species and their close relatives are discussed. Hypomyces orthosporus is found for the first time from China.

The genera Rugonectria and Thelonectria (Hypocreales, Nectriaceae) in China.

Recent collections and herbarium specimens of Rugonectria and Thelonectria from different regions of China were examined. Using combined analyses of morphological and molecular data, 17 species are recognised including three species of Rugonectria and 14 species in Thelonectria. Amongst them, R.microconidia and T.guangdongensis are new to science. Rugonectriamicroconidia on mossy bark is characterised by superficial, yellow to orange, pyriform to subglobose perithecia with a warted surface; ellipsoidal to broadly ellipsoidal, striate, uniseptate ascospores; and allantoid to rod-shaped, aseptate microconidia. Thelonectriaguangdongensis possesses bright red perithecia with a slightly roughened surface and a prominently dark papilla; ellipsoidal, smooth, uniseptate ascospores; and subcylindrical, slightly curved, multiseptate macroconidia. Morphological distinctions and sequence divergences between the new species and their close relatives are discussed. Name changes for the previously recorded species in China are noted.

Two new species of Geejayessia (Hypocreales) from Asia as evidenced by morphology and multi-gene analyses.

Two new species of Geejayessia are introduced, based on materials collected from central China. Geejayessiaclavata sp. nov. is characterised by gregarious, red brownish to dark red, oval-subglobose to globose perithecia that are formed on a basal stroma; (4-7-)8-spored cylindrical asci; ellipsoidal or rarely broadly ellipsoidal, uniseptate, smooth or finely verruculose ascospores; clavate, aseptate microconidia and absence of macroconidia. Geejayessiasinica sp. nov. is characterised by red to bright red, pyriform, subglobose to globose, perithecia on a basal stroma, collapsing laterally when dry; subcylindrical to clavate asci with a rounded apex; ellipsoidal, uniseptate ascospores; and falcate, multiseptate macroconidia with an arcuate tip. Morphological distinctions of the new species from the related fungi are discussed. This is the first report of Geejayessia from Asia.

The complete mitochondrial genome of the bambusicolous fungus Fusarium bambusae (Nectriaceae, Ascomycota).

The complete nucleotide sequence of the mitochondrial genome of the bambusicolous fungus Fusarium bambusae was determined using the next-generation sequencing technology. The circular molecule is 63,593 bp long with a GC content of 31.92%. Gene prediction revealed 44 genes encoding 15 conserved proteins, 27 tRNAs, and the large and small ribosomal RNAs. All genes are located on the same strand. The tRNA genes contain codons for all 20 standard amino acids. It turns out to be similar to the previously sequenced mitochondrial genomes of Fusarium circinatum and F. verticillioides. The differences lie in the number of introns embodied in protein-coding genes. Four introns exist in the mitochondrial genome of F. verticillioides, 10 in F. bambusae, and 14 in F. circinatum. The phylogenetic analysis confirmed F. bambusae as a member of Fusarium (Nectriaceae). The mitochondrial genome of F. bambusae will contribute to the understanding of phylogeny and evolution of the genus and family.

Phylogeny and morphological analyses of Penicillium section Sclerotiora (Fungi) lead to the discovery of five new species.

Phylogeny of Penicillium section Sclerotiora is still limitedly investigated. In this study, five new species of Penicillium are identified from the samples collected from different places of China, and named P. austrosinicum, P. choerospondiatis, P. exsudans, P. sanshaense and P. verrucisporum. The conidiophores of P. austrosinicum and P. exsudans are monoverticillate like most members of the section, while the rest species are biverticillate similar to the only two species P. herquei and P. malachiteum previously reported in the section Sclerotiora. The phylogenetic positions of the new taxa are determined based on the sequence data of ITS, BenA, CaM and RPB2 regions, which reveals that all the species with biverticillate condiophores form a well-supported subclade in the section. The new Penicillium species clearly differ from the existing species of the genus in culture characteristics on four standard growth media, microscopic features, and sequence data. Morphological discrepancies are discussed between the new species and their allies.

The complete mitochondrial genome of the important mycoparasite Clonostachys rosea (Hypocreales, Ascomycota).

The complete nucleotide sequence of mitochondrial genome of the important mycoparasitic fungus Clonostachys rosea was determined using the next-generation sequencing technology. The circular molecule is 40,921 bp long with a GC content of 27.90%. Gene prediction revealed 42 genes encoding 15 conserved proteins, 25 tRNAs, the large and small ribosomal RNAs. All genes are located on the same strand. It is found to be similar to the previously sequenced mitochondrial genomes of Acremonium chrysogenum and Nectria cinnabarina. The differences lie in the copy number of trnG-UCC and locations of trnN-GUU and cox2. The phylogenetic analysis confirmed C. rosea as a sister taxon of A. chrysogenumin (Bionectriaceae). The mitochondrial genome of C. rosea will contribute to the understanding of phylogeny and evolution of Hypocreales.

The complete mitochondrial genome of the important phytopathogen Nectria cinnabarina (Hypocreales, Ascomycota).

The complete nucleotide sequence of the mitochondrial genome of the important phytopathogic fungus Nectria cinnabarina was determined using the next-generation sequencing technology. The circular molecule is 69 895 bp long with a GC content of 28.71%. Gene prediction revealed 42 genes encoding 15 conserved proteins, 25 tRNAs, the large and small ribosomal RNAs. All genes are located on the same strand. Compared with previously sequenced mitochondrial genomes of the other members of Nectriaceae, the composition and order of the protein and rRNA genes are highly conserved; however, the quantity and order of tRNA genes are different. The phylogenetic analysis confirmed N. cinnabarina as a basal lineage in Nectriaceae. The mitochondrial genome of N. cinnabarina will contribute to the understanding of phylogeny and evolution of Nectriaceae and Hypocreales.

Expression of interleukin-17 associated with disease progression and liver fibrosis with hepatitis B virus infection: IL-17 in HBV infection.

BACKGROUND/OBJECTIVES: As a proinflammatory cytokine, interleukin-17 (IL-17) contributes to the inflammation of many autoimmune diseases. We examined IL-17 levels in serum and tissues from patients with chronic hepatitis B virus infection (HBV), and especially evaluated the role of IL-17 in the pathogenesis and progression of liver fibrosis. MATERIALS AND METHODS: Whole venous blood was obtained from four patient groups: chronic hepatitis B (CHB, n = 47), liver cirrhosis (LC, n = 49), primary hepatocellular carcinoma (PHC, n = 44), chronic liver failure (CLF, n = 33), and a normal control group (n = 20). HBsAg was positive in all patients. Liver biopsy samples were acquired from asymptomatic HBsAg carriers (ASC, n = 35), CHB (n = 57), and LC (n = 31) patients. We performed ELISA to measure IL-17 levels in serum samples, and used reverse RT-PCR to measure IL-17 mRNA levels in peripheral blood mononuclear cells (PBMC). IL-17 protein expression was detected in liver biopsy tissues by immunohistochemistry. RESULTS: Compared to normal controls, serum IL-17 protein and mRNA levels were significantly higher in the four infection groups. LC patients exhibited the highest serum IL-17 and PBMC mRNA levels. No significant differences were found between the other three groups. High levels of IL-17 were also observed in tissues from CHB and LC patients, compared to ASC. IL-17 expression was mainly located in the portal area and was positively correlated with inflammation grade and fibrosis stage. CONCLUSIONS: IL-17 expression was found to be increased with increasing degrees of liver fibrosis. This suggests that IL-17 may not only induce the inflammation, but also contribute to disease progression and chronicity. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5306959258322482.

[Comparison of genomes between Aspergillus nidulans and 30 filamentous ascomycetes].

To investigate the conserved homologs of filamentous ascomycetes genomes, the local fungal genome database used in this analysis was established, which consisted of 31 latest and complete genome data publicly available on the Internet. An expectation value cutoff of 0.1 was used to identify significant hits. Each complete gene set of the query genome Aspergillus nidulans genome with 10,560 annotated genes was splitted into individual FASTA files with Seqverter and then compared separately against each filamentous ascomycete genome using Standalone BLASTN. The result indicated that the number of matches reflected the evolutional relationships of the filamentous ascomycetes analysed. Of 10,560 genes in Aspergillus nidulans genome, 924 had match sequences with other 30 filamentous ascomycetes ones. The number of homology sequences were 6, 3, 6, and 6 at E-values in the range of 10(-5) to 0.1, 10(-30) to 10(-5), 10(-100) to 10(-30) and 0 to 1000(-100), respectively. Six homologs at E-values ranging from 10(-5) to 0.1 and 3 at E-values ranging from 10(-30) to 10(-5) were variable, while the 6 at E-values ranging from 0 to 10(-100) were highly conserved based on the alignments using ClustalX. Six homologs were relatively conserved at E-values in the range of 10(-100) to 10(-30), which can be used in phylogeny of these filamentous ascomycetes in this study.