Bacillus velezensis BV01 Has Broad-Spectrum Biocontrol Potential and the Ability to Promote Plant Growth.

To evaluate the potential of a bacterial strain as a fungal disease control agent and plant growth promoter, its inhibitory effects on phytopathogens such as Bipolaris sorokiniana, Botrytis cinerea, Colletotrichum capsici, Fusarium graminearum, F. oxysporum, Neocosmospora rubicola, Rhizoctonia solani, and Verticillium dahliae were investigated. The results showed that the inhibitory rates in dual-culture and sterile filtrate assays against these eight phytopathogens ranged from 57% to 83% and from 36% to 92%. The strain was identified as Bacillus velezensis based on morphological and physiological characterization as well as phylogenetic analyses of 16S rRNA and the gyrase subunit A protein (gyrA) regions. The results demonstrated that B. velezensis was able to produce fungal cell-wall-degrading enzymes, namely, protease, cellulase, and ß-1,3-glucanase, and the growth-promotion substances indole-3-acetic acid (IAA) and siderophore. Furthermore, B. velezensis BV01 had significant control effects on wheat root rot and pepper Fusarium wilt in a greenhouse. Potted growth-promotion experiments displayed that BV01 significantly increased the height, stem diameter, and aboveground fresh and dry weights of wheat and pepper. The results imply that B. velezensis BV01, a broad-spectrum biocontrol bacterium, is worth further investigation regarding its practical applications in agriculture.

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.

First Report of Cobweb Disease in Auricularia cornea var. Li. Caused by Hypomyces mycophilus in China.

Auricularia cornea var. Li. is a natural mutation strain of A. cornea which has been preferred by consumers for its white colour, good taste and pharmacological properties (Rebecca et al. 2020). In October 2021, a disease with symptoms similar to that of cobweb disease (Carrasco et al. 2017) was observed in A. cornea var. Li. in four mushroom farms in Fangshan District (115.83°E, 39.55°N), Beijing, China, infecting 20% of the fruiting bodies (Fig. 1A-D). White cottony mycelia formed typically on the casing soil and they gradually spread to the stipes and pileus, covering the whole fruiting body, which eventually died and lost commodity value. Cultures were obtained by aseptically transferring the diseased fruiting bodies onto potato dextrose agar (PDA); they were deposited in the culture collection (ID: JZBQA3) of the Beijing Academy of Agriculture and Forestry Sciences, China. The colonies were floccose with aerial mycelium white. Purplish grey diffusing pigments occasionally formed on the reverse side of the plate at 25 °C (Fig. 2A-B). Conidiophores arising in aerial mycelium, indefinite in length, branches septate, each cell producing denticulate conidiogenous loci, each denticle bearing a single conidium. Conidia mostly oblong to ellipsoidal, smooth, (9.0-)9.9-17.0(-18.0)×(6.0-)6.9-10.2 µm (n = 60), 0~1 septate (Fig. 2C-E). Chlamydospores forming as lateral branches of hyphae were commonly observed, globose, ellipsoid or oblong, 14.8-22×14.7-19.6 µm, l/w = 1.0-1.3 (Fig. 2F-G). The morphological characteristics were consistent with that of Hypomyces mycophilus, whose anamorph was Cladobotryum polypori (Rogerson et al. 1993, Zeng et al. 2017). For further species identification, genomic DNA was extracted using the DNeasy Plant Mini Kit (Qiagen, USA). The internal transcribed spacer (ITS) regions and large subunit ribosomal RNA (LSU) genes were amplified using the primer pairs ITS1/ITS4 (White et al. 1990) and LROR/LR5 (Bhattacharya et al. 2000), respectively. The sequences were deposited in GenBank with accession numbers OP430530 and OP430531. BLAST nucleotide searches showed more than 99% homology with corresponding sequences of Hypomyces mycophilus HMAS 275554 and CBS 175.56. Phylogenetic trees based on ITS and LSU revealed that the strain JZBQA3 was grouped with H. mycophilus with high support value (Fig. 3). A in vivo pathogenicity test was performed using eight mushroom sticks with healthy fruiting bodies in triplicate. Each four sticks were sprayed with conidial suspension (108 spores/mL) of strain JZBQA3 or sterile distilled water, respectively, and maintained in an artificial climate chamber at 25-26°C. Cobweb-like characteristics were observed on the fruiting bodies treated with the JZBQA3 conidial suspension 2-3 days after inoculation, while those treated with sterile distilled water remained symptomless (Fig. 4 A-B). The same pathogen was re-isolated and confirmed from the infected fruiting bodies by integrated analysis of morphological characteristics and gene sequencing data, fulfilling Koch's postulates. Hypomyces mycophilus was first reported on Trametes versicolor in North Carolina (Rogerson et al. 1993), and is the causal agent of cobweb diseases on Auricularia heimuer (Zhang et al. 2023). To our knowledge, this is the first report of cobweb disease caused by H. mycophilus in A. cornea var. Li. This finding is a valuable contribution to the knowledge of cobweb disease development in edible fungi.

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.

First Report of Cobweb Disease in Oudemansiella raphanipes Caused by Cladobotryum varium in Beijing, China.

Oudemansiella raphanipes is an edible mushroom with medicinal properties,which has been recently cultivated throughout China (Hao et al. 2016). In October 2019, a disease with symptoms similar to that of cobweb disease (Carrasco et al. 2017) was observed in O. raphanipes in the Tongzhou District, Beijing, China, infecting 25% of the fruiting bodies (Fig. 1A, B). White cotton-like net of hyphae were present typically on the casing soil or on the stipe of the fruiting bodies; they gradually spread to the pileus, covering the fruiting body, which eventually wilted and died (Fig. 1C, D), resulting in yield reduction and economic loss. Cultures were obtained by aseptically transferring the diseased fruiting bodies onto potato dextrose agar (PDA) at 25 °C; they were deposited in the culture collection (ID: JZBQA1) of the Beijing Academy of Agricultural and Forestry Sciences, China. The colonies were pale white/white, with an occasional formation of yellow diffusing pigments on the reverse side (Fig. 1E-G). Conidiophores were Cladobotryum-like, phialides were solitary or commonly divergent in whorls of 2-3 (-4), lageniform to subulate, 20-63.5 (-66) × (3.8-) 4-5.3 (-9) µm (n = 40) (Fig. 1H, I); conidia were hyaline, oval to ellipsoidal, with one or two septa, (10.4-) 11.4-20 (-22) × 6.6-9.5 (-10) µm (n = 40) (Fig. 1J); chlamydospores were globose or ellipsoidal (Fig. 1K). The morphological characteristics were consistent with that of Cladobotryum varium (Back et al. 2012a, b; Sun et al. 2019). For species-level fungal identification, genomic DNA was extracted using the DNeasy Plant Mini Kit (Qiagen, USA). The internal transcribed spacer (ITS) regions, translation elongation factor 1 alpha exon (TEF1-α), RNA polymerase II subunit b (RPB2), and RNA polymerase I largest subunit (RPB1) genes were amplified using the primer pairs ITS1/ITS4 (White et al. 1990), EF1-983F/2218R (Rehner and Buckley 2005), RPB2-5F/7cR (Liu et al. 1999), and RPB1F1 (5'-GCCGATGAAGTTGGTCTA-3')/RPB1R1 (5'-TATGTTGCGGTGAGCCTT-3'), respectively. A BLAST nucleotide search showed 99.34% (449/452 bp), 99.24% (914/921 bp), 98.08% (1,022/1,042 bp), and 99.66% (588/590 bp) homology, respectively, with those of the ex-type culture of Hypomyces aurantius TFC 95-171 (FN859425.1, FN868743.1, FN868679.1, and FN868805.1). The four sequences were deposited in GenBank (accession numbers: MW534093, MW560066, MW560064, and MW560065). Phylogenetic trees based on the assessed gene loci revealed that the JZBQA1 strain was closely related to C. varium (Fig. 2). A in vivo pathogenicity test was performed using the fruiting bodies (Fig. 1L, O). Spore suspension (108 spores/mL) of the JZBQA1 strain or sterile distilled water was sprayed on six healthy fruiting bodies, maintained in an artificial climate chamber at 24-26°C. Cobweb-like features were observed on the fruiting bodies treated with the spore suspension 2-3 days post-inoculation; while those treated with water did not exhibit such features (Fig. 1L, O). The same pathogen was re-isolated and confirmed from the infected fruiting bodies by integrated analysis of morphological characteristics and gene sequencing data. Cladobotryum spp. infects different varieties of cultivated edible mushrooms, resulting in the development of cobweb diseases (Cao et al. 2020; Carrasco et al. 2017). Cladobotryum varium is the causal agent of cobweb disease in Flammulina velutipes and Hypsizygus marmoreus (Back et al. 2012a, b). To our knowledge, this is the first report of cobweb disease caused by C. varium in O. raphanipes. This finding is a valuable contribution to the knowledge of cobweb disease development in edible fungi.

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.

Analysis of the Anti-Inflammatory and Analgesic Mechanism of Shiyifang Vinum Based on Network Pharmacology.

OBJECTIVE: The possible core active compounds and potential mechanism of action of Shiyifang Vinum were explored through network pharmacology and in vitro enzyme activity verification experiments. METHODS: We screened the core active components and the action targets of Shiyifang Vinum through the TCMSP database and literature mining and drew a Venn map of the intersection with anti-inflammatory and analgesic-related gene targets. Go and KEGG analyses were enriched with the David database. The compound target pathway network was constructed using Cytoscape 3.6.1. The binding strength of core active compounds and target proteins was verified through molecular docking, and the direct effects of Shiyifang Vinum and four monomer compounds on COX-2 enzyme activity were detected through an in vitro enzyme activity test. RESULTS: 14 active compounds and 11 targets were screened out from Shiyifang Vinum through TCMSP database and literature mining; 252 GO entries were obtained by GO analysis, and 114 signal pathways were screened by KEGG analysis. The results of the molecular docking showed that the core compounds and target proteins had strong binding activity. In vitro validation experiments showed that both the Shiyifang Vinum and the four monomer compounds could inhibit the activity of COX-2. CONCLUSION: This study preliminarily explored the potential active compounds and target proteins of the anti-inflammatory and analgesic effects of Shiyifang Vinum, which could provide a scientific basis for further study on the anti-inflammatory and analgesic mechanism and material basis of this recipe.

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.

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.

Selection of a DNA barcode for Nectriaceae from fungal whole-genomes.

A DNA barcode is a short segment of sequence that is able to distinguish species. A barcode must ideally contain enough variation to distinguish every individual species and be easily obtained. Fungi of Nectriaceae are economically important and show high species diversity. To establish a standard DNA barcode for this group of fungi, the genomes of Neurospora crassa and 30 other filamentous fungi were compared. The expect value was treated as a criterion to recognize homologous sequences. Four candidate markers, Hsp90, AAC, CDC48, and EF3, were tested for their feasibility as barcodes in the identification of 34 well-established species belonging to 13 genera of Nectriaceae. Two hundred and fifteen sequences were analyzed. Intra- and inter-specific variations and the success rate of PCR amplification and sequencing were considered as important criteria for estimation of the candidate markers. Ultimately, the partial EF3 gene met the requirements for a good DNA barcode: No overlap was found between the intra- and inter-specific pairwise distances. The smallest inter-specific distance of EF3 gene was 3.19%, while the largest intra-specific distance was 1.79%. In addition, there was a high success rate in PCR and sequencing for this gene (96.3%). CDC48 showed sufficiently high sequence variation among species, but the PCR and sequencing success rate was 84% using a single pair of primers. Although the Hsp90 and AAC genes had higher PCR and sequencing success rates (96.3% and 97.5%, respectively), overlapping occurred between the intra- and inter-specific variations, which could lead to misidentification. Therefore, we propose the EF3 gene as a possible DNA barcode for the nectriaceous fungi.