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The application of arylacetonitrilases from filamentous fungi to the hydrolysis of high concentrations of (R,S)-mandelonitrile (100-500 mM) was demonstrated for the first time. Escherichia coli strains expressing the corresponding genes were used as whole-cell catalysts. Nitrilases from Aspergillus niger, Neurospora crassa, Nectria haematococca, and Arthroderma benhamiae (enzymes NitAn, NitNc, NitNh, and NitAb, respectively) exhibited different degrees of enantio- and chemoselectivity (amide formation). Their enantio- and chemoselectivity was increased by increasing pH (from 8 to 9-10) and adding 4-10% (v/v) toluene as the cosolvent. NitAn and NitNc were able to convert an up to 500 mM substrate in batch mode. NitAn formed a very low amount of the by-product, amide (1% of the total product). This enzyme produced up to70 g/L of (R)-mandelic acid (e.e. 94.5-95.6%) in batch or fed-batch mode. Its volumetric productivities were the highest in batch mode [571 ± 32 g/(L d)] and its catalyst productivities in fed-batch mode (39.9 ± 2.5 g/g of dcw). NitAb hydrolyzed both enantiomers of 100 mM (R,S)-mandelonitrile at pH 5.0 and is therefore promising for the enantioretentive transformation of (S)-mandelonitrile. Sequence analysis suggested that fungal arylacetonitrilases with similar properties (enantioselectivity, chemoselectivity) were clustered together.

Based on type studies and freshly collected material we here re-instate the genus Thyronectria (Nectriaceae, Hypocreales). Species of this genus were recently for the most part classified in the genera Pleonectria (Nectriaceae) or Mattirolia (Thyridiaceae), because Thyronectria and other genera had been identified as members of the Thyridiaceae due to the presence of paraphyses. Molecular phylogenies based on several markers (act, ITS, LSU rDNA, rpb1, rpb2, tef1, tub) revealed that the Nectriaceae contain members whose ascomata are characterised by long, more or less persistent, apical paraphyses. All of these belong to a single genus, Thyronectria, which thus has representatives with hyaline, rosy, green or even dark brown and sometimes distoseptate ascospores. The type species of Thyronectria, T. rhodochlora, syn. T. patavina, syn. T. pyrrhochlora is re-described and illustrated. Within the Nectriaceae persistent, apical paraphyses are common in Thyronectria and rarely also occur in Nectria. The genus Mattirolia is revised and merged with Thyronectria and also Thyronectroidea is regarded as a synonym of Thyronectria. The three new species T. asturiensis, T. caudata and T. obscura are added to the genus. Species recently described in Pleonectria as well as some species of Mattirolia are combined in the genus, and a key to Thyronectria is provided. Five species are epitypified. The type species of the genus Thyridium (Thyridiaceae), T. vestitum, is included in phylogenetic analyses to illustrate the phylogenetic distance of Thyronectria from the Thyridiaceae.

Compared to (S)-selective amine transaminase ((S)-AT), the (R)-selective counterpart ((R)-AT) has been less studied. As such, a simplified "Motif Sequence Blast" search (Höhne et al. Nat Chem Biol 6:807-813, 2010) was carried out to identify new (R)-ATs from the protein databases. The combined conserved sequence motifs of (R)-ATs based on the previous in silico method of predicting (R)-selective amine transaminase were used as the template sequence for BLASTP search at default settings in NCBI, and six candidate sequences were identified. These putative (R)-AT genes were synthesized and overexpressed in Escherichia coli. Among them, five new (R)-ATs were expressed as soluble protein and showed unusual substrate specificity and high stereoselectivity. Furthermore, several unnatural amino acids, such as D-alanine, D-2-aminobutyric acid, and D-norvaline, were synthesized via the (R)-AT-catalyzed amino transfer reaction to the corresponding keto acids. Optically pure (S)-amines were also obtained by kinetic resolution of racemic amines catalyzed with these new (R)-ATs. Therefore, the Motif Sequence Blast search offers a quick and effective method for in silico identification of new (R)-ATs, and the newly identified (R)-ATs are attractive additions to the toolbox of (R)-ATs for further study and industrial application.

To our best knowledge, all of the fungal immunomodulatory proteins (FIPs) have been successfully extracted and identified in Basidomycetes, with only the exception of FIP from ascomycete Nectria haematococca (FIP-nha) discovered through homology alignment most recently. In this work, a gene encoding FIP-nha was synthesized and recombinantly expressed in an Escherichia coli expression system. SDS-PAGE and MALDI-MS analyses of recombinant FIP-nha (rFIP-nha) indicated that the gene was successfully expressed. The yield of the bioactive FIP-nha protein was 42.7 mg/L. In vitro assays of biological activity indicated that the rFIP-nha caused hemagglutination of human and rabbit red blood cells, significantly stimulated mouse spleen lymphocyte proliferation, and enhanced expression of interleukin-2 (IL-2) released from mouse splenocytes, revealing a strong antitumor effect against HL60, HepG2 and MGC823. Through this work, we constructed a rapid and efficient method of FIP production, and suggested that FIP-nha is a valuable candidate for use in future medical care and pharmaceutical products.

Nineteen species of lichenicolous fungi collected in 129 localities in Poland in the years 1968 and 1970-2003 are reported in the paper. They are as follows: Athelia arachnoidea (Berk.) Jülich, Tremella cladoniae Diederich et M.S. Chrst., T. hypogymniae Diederich et M.S. Chris., T. lichecola Diederich, Clypeococcum hypocenomycis D. Hawksw., Polycoccum superficiale D. Hawksw. et Miądlikowska, Nectria lecanodes Ces., Pronectria erythrinella (Nyl.) Lowen, Cortocifraga fuckelii (Rehm) D. Hawksw. et R. Sant., C. peltigerae (Nyl.) D. Hawksw. et R. Sant., Libertiella malmedyensis Speg. et Roum., Lichenoconium erodens M.S. Christ. et D. Hawksw., L. lecanorae (Jaap) D. Hawksw., L. pyxidatae (Oudem.) Petrak et Sydow, Vouauxiella lichenicola (Lindsay) Petrak et Sydow, Bispora christiansenii D. Hawksw., Illosporium carneum Fr., Karsteniomyces peltigerae (P. Karst.) D. Hawksw. and Taeniolella beschiana Diederich.

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

Oxalate film formation is a pathology that often occurs in mural paintings and may result from the concomitant action of microorganisms and environmental conditions. Low Choir of the Convent of Nossa Senhora da Saudacao (Portugal) has mural paintings with an extraordinary beauty, which over time have been suffered polychromy degradation and biofilm formation, presenting an ideal case study to investigate the role and impact of microorganisms in the biodeterioration process. Bacterial populations, filamentous fungi belonging to the genera Cladosporium, Penicillium , Nectria and yeast strain of the genera Rhodotorula were isolated from these wall paintings. The penetration of fungal hyphae in the microstructure of mortars, observed by scanning electron microscopy, seems to be responsible for cracking and detachments in some areas of the painting. The study revealed that the veils on the surface of the paintings are essentially oxalates and that these biofilms are caused by metabolic activity of bacterial communities. Furthermore, the colour alteration of green areas due to microorganisms was detected by Raman microscopy, in real samples and under in vitro conditions, being the result of the metabolic activity of microorganisms present on the paintings, which promote calcium oxalates formation over the malachite paint layers.

Introduction. Keratinophilic fungi are an important group of fungi that live in soil. The aim of this study was to isolate and identify keratinophilic fungi from the soil of different parks in Shiraz.Materials and Methods. A total of 196 soil samples from 43 parks were collected. Isolation of the fungi was performed by hair bait technique. The isolated colonies were identified by morphologic feature of macro- and microconidia and molecular method, using DNA sequence analysis. ITS region of ribosomal DNA was amplified and the PCR products were sequenced.Results. 411 isolates from 22 genera were identified.Fusarium(23.8%),Chrysosporium(13.13%),Acremonium(12.65%),Penicillium(12.39%),Microsporum gypseum(1.94%),Bionectria ochroleuca(1.21%),Bipolaris spicifera(1.21%),Scedosporium apiospermum(0.82%),Phialophora reptans(0.82%),Cephalosporium curtipes(0.49%),Scedosporium dehoogii(0.24%),Ochroconis constricta(0.24%),Nectria mauritiicola(0.49%),Chaetomium(0.49%),Scopulariopsis(0.24%),Malbranchea(0.24%), andTritirachium(0.24%) were the most important isolates. Most of the fungi were isolated from the soils with the PH range of 7 to 8.Conclusion. Our study results showed that many keratinophilic fungi isolated from the parks soil are important for public health and children are an important group at a high risk of being exposed to these fungi.

A new genus, Xanthonectria, is proposed for the enigmatic taxon Nectria pseudopeziza. Xanthonectria is characterized by nonstromatic, soft-textured, superficial, glabrous, pale yellow to bright orange nectria-like ascomata, not changing color in 3% KOH or lactic acid, long fusiform, multiseptate ascospores and an acremonium-like asexual morph. Its placement in the Bionectriaceae and its relationship to other members of this family are based on morphological, cultural and molecular data

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.

Se evaluaron posibles métodos de control biológico a través de microorganismos antagonistas, por medio de pruebas en laboratorio en las cuales se puso a competir el organismo antagonista contra el agente patógeno causal de la enfermedad denominada “Nectria”, el cual fue determinado en éste mismo estudio. Adicionalmente, se realizaron pruebas de inoculación en vivero, así como ensayos en plantaciones forestales propiedad de la empresa Ethical Forestry S.A. en la Zona Norte de Costa Rica. Los resultados determinaron que el principal agente patógeno de la enfermedad es Fusarium solani. Las pruebas de laboratorio en las cuales se confrontaron los microorganismos antagonistas contra el patógeno, demostraron que tanto Trichoderma harzianum como Trichoderma asperellum generan resultados acertados en el control del patógeno, no obstante se reporta un crecimiento más agresivo de T. harzianum. A nivel de vivero, se obtuvieron diferencias importantes en los porcentajes de germinación en los tratamientos de T. harzianum más T. asperellum (52% superior) y Bacillus subtilis, Pseudomonas fluorescens, Azotobacter chroococcum (21% superior) respecto al tratamiento testigo; en cuanto a ensayos de campo la incidencia de la enfermedad en plantaciones de 21 meses de edad ha sido prácticamente nula, además a pesar de no encontrar diferencias estadísticamente significativas se ha podido notar un crecimiento superior tanto en diámetro (hasta 1,15 cm en promedio) como en altura (hasta 1,03 metros en promedio) en los tratamientos a base de microorganismos antagonistas.

This study led to the discovery of four putative entomopathogenic fungi of armoured scale insects on citrus trees in coastal New South Wales. Two of these species belong in Podonectria as P. coccicola (Ellis & Everh.) Petch (syn. Tetracrium coccicola (Hohn.) Ellis & Everh.) and P. novae-zelandiae Dingley. Members of this genus are grown in culture for the first time. Formerly placed in the Pleosporales, Tubeufiaceae, or more recently in the Tubeufiales, these species are herein placed in the new family Podonectriaceae fam. nov., Pleosporales. Another species is placed in the Hypocreales, Bionectriaceae as Clonostachys coccicola (J.A. Stev.) H.T. Dao comb. nov. (basionym Tubercularia coccicola J.A. Stev., syn. Nectria tuberculariae Petch). The fourth species is Myriangium citri Henn. (Myriangiales, Myriangiaceae). Each fungal species is characterized and the phylogenetic placement confirmed by molecular analyses of the ITS and 28 s rDNA regions. In addition, their biology is noted, including location of the fungi within tree canopies.

Four new polyketides: nectriacids A–C (1–3) and 12-epicitreoisocoumarinol (4), together with three known compounds: citreoisocoumarinol (5), citreoisocoumarin (6), and macrocarpon C (7) were isolated from the culture of the endophytic fungus Nectria sp. HN001, which was isolated from a fresh branch of the mangrove plant Sonneratia ovata collected from the South China Sea. Their structures were determined by the detailed analysis of NMR and mass spectroscopic data. The absolute configuration of the stereogenic carbons for compound 4 was further assigned by Mosher’s ester method. All of the isolated compounds were tested for their α-glucosidase inhibitory activity by UV absorbance at 405 nm, and new compounds 2 and 3 exhibited potent inhibitory activity with IC50 values of 23.5 and 42.3 μM, respectively, which were more potent than positive control (acarbose, IC50, 815.3 μM).

Host specificity of plant pathogens can be dictated by genes that enable pathogens to circumvent host defenses. Upon recognition of a pathogen, plants initiate defense responses that can include the production of antimicrobial compounds such as phytoalexins. The pea pathogen Nectria haematococca mating population VI (MPVI) is a filamentous ascomycete that contains a cluster of genes known as the pea pathogenicity (PEP) cluster in which the pisatin demethylase (PDA) gene resides. The PDA gene product is responsible for the detoxification of the phytoalexin pisatin, which is produced by the pea plant (Pisum sativum L.). This detoxification activity allows the pathogen to evade the phytoalexin defense mechanism. It has been proposed that the evolution of PDA and the PEP cluster reflects horizontal gene transfer (HGT). Previous observations consistent with this hypothesis include the location of the PEP cluster and PDA gene on a dispensable portion of the genome (a supernumerary chromosome), a phylogenetically discontinuous distribution of the cluster among closely related species, and a bias in G+C content and codon usage compared to other regions of the genome. In this study we compared the phylogenetic history of PDA, beta-tubulin, and translation elongation factor 1-alpha in three closely related fungi (Nectria haematococca, Fusarium oxysporum, and Neocosmospora species) to formally evaluate hypotheses regarding the origin and evolution of PDA. Our results, coupled with previous work, robustly demonstrate discordance between the gene genealogy of PDA and the organismal phylogeny of these species, and illustrate how HGT of pathogenicity genes can contribute to the expansion of host specificity in plant-pathogenic fungi.

During the nine years (2002-2010) 6 apple cultivars, scab-resistant or partially resistant to apple scab: ‘Bohemia’, ‘Topaz’, ‘Rubinola’, ‘Goldstar’, ‘Pinova’ and ‘Redkroft’ grafted on 3 dwarfing rootstock: M.9, P 22 and P 60, were tested at the Experimental Orchard in Dąbrowice with integrated fruit production. The susceptibility to apple canker (Nectria sp.), apple scab and powdery mildew, as well as growth, yielding, fruit quality and damage by pests were observed. After 9 years, the highest number of infected trees, on trunks and branches, were observed on the cultivars: ‘Topaz’, ‘Bohemia’, and ‘Redkroft’. All the cultivars grafted on P 60 rootstocks formed the largest trees, whereas on P 22 the smallest. The highest yields were obtained from all the cultivars grown on P 60 and the smallest on P 22. The cropping efficiency index was the highest for: ‘Bohemia’, ‘Rubinola’, ‘Goldstar’ and ‘Pinova’ on P 22. ‘Topaz’ had the highest cropping efficiency on P 60 and ‘Redkroft’ on M.9 and P 60. In the last three years (2008-2010), the effect of rootstock on fruit quality was minimal and not significant. Apple scab and powdery mildew, and some important pests, were not observed after applying on integrated system of spraying. The apple cultivars: ‘Bohemia’, ‘Rubinola’, ‘Topaz ‘and ‘Redkroft’ are the most promising for processing and amateur orchards with the integrated system of management. The cultivars: ‘Pinova’ and ‘Goldstar’ should not be recommended for growing on light soil in the integrated system of production. The fruits of ‘Goldstar’ are susceptible to bitter pit.

Although Nectria is the type genus of Nectriaceae (Hypocreales, Sordariomycetes, Pezizomycotina, Ascomycota), the systematics of the teleomorphic and anamorphic state of Nectria sensu Rossman has not been studied in detail. The objectives of this study are to 1) provide a phylogenetic overview to determine if species of Nectria with Gyrostroma, Tubercularia, and Zythiostroma anamorphs form a monophyletic group; 2) define Nectria, segregate genera, and their species using morphologically informative characters of teleomorphic and anamorphic states; and 3) provide descriptions and illustrations of these genera and species. To accomplish these objectives, results of phylogenetic analyses of DNA sequence data from six loci (act, ITS, LSU, rpb1, tef1 and tub), were integrated with morphological characterisations of anamorphs and teleomorphs. Results from the phylogenetic analyses demonstrate that species previously regarded as the genus Nectria having Gyrostroma, Tubercularia, and Zythiostroma anamorphs belong in two major paraphyletic clades. The first major clade regarded as the genus Pleonectria contains 26 species with ascoconidia produced by ascospores in asci, perithecial walls having bright yellow scurf, and immersed or superficial pycnidial anamorphs (Zythiostroma = Gyrostroma). A lineage basal to the Pleonectria clade includes Nectria miltina having very small, aseptate ascospores, and trichoderma-like conidiophores and occurring on monocotyledonous plants. These characteristics are unusual in Pleonectria, thus we recognise the monotypic genus Allantonectria with Allantonectria miltina. The second major clade comprises the genus Nectria sensu stricto including the type species, N. cinnabarina, and 28 additional species. Within the genus Nectria, four subclades exist. One subclade includes species with sporodochial anamorphs and another with synnematous anamorphs. The other two paraphyletic subclades include species that produce abundant stromata in which the large perithecia are immersed, large ascospores, and peculiar anamorphs that form pycnidia or sporodochia either on their natural substrate or in culture. In this study the evolution of species, morphology, and ecology of the three genera, Allantonectria, Nectria, and Pleonectria, are discussed based on the phylogenetic analyses. In addition, descriptions, illustrations, and keys for identification are presented for the 56 species in Allantonectria, Nectria, and Pleonectria. Taxonomic novelties: New species: Nectria argentinensis Hirooka, Rossman & P. Chaverri, Nectria berberidicola Hirooka, Lechat, Rossman, & P. Chaverri, Nectria himalayensis Hirooka, Rossman, & P. Chaverri, Nectria magnispora Hirooka, Rossman, & P. Chaverri, Nectria mariae Hirooka, Fournier, Lechat, Rossman, & P. Chaverri, Nectria pyriformis Hirooka, Rossman & P. Chaverri, Pleonectria boothii Hirooka, Rossman & Chaverri, Pleonectria clavatispora Hirooka, Rossman & P. Chaverri, Pleonectria ilicicola Hirooka, Rossman & P. Chaverri, Pleonectria okinawensis Hirooka, Rossman & P. Chaverri, Pleonectria pseudomissouriensis Hirooka, Rossman & P. Chaverri, Pleonectria quercicola Hirooka, Checa, Areual, Rossman & P. Chaverri, Pleonectria strobi Hirooka, Rossman & P. Chaverri. New combinations: Cosmospora proteae (Marinc., M.J. Wingf. & Crous) Hirooka, Rossman & P. Chaverri, Nectricladiella viticola (Berk. & M.A. Curtis) Hirooka, Rossman & P. Chaverri, Neocosmospora guarapiensis (Speg.) Hirooka, Samuels, Rossman & P. Chaverri, Neocosmospora rehmiana (Kirschstein) Hirooka, Samuels, Rossman & P. Chaverri, Pleonectria aquifolii (Fr.) Hirooka, Rossman & P. Chaverri, Pleonectria aurigera (Berk. & Rav.) Hirooka, Rossman & P. Chaverri, Pleonectria chlorinella (Cooke) Hirooka, Rossman & P. Chaverri, Pleonectria coryli (Fuckel) Hirooka, Rossman & P. Chaverri, Pleonectria cucurbitula (Tode: Fr.) Hirooka, Rossman & P. Chaverri, Pleonectria lonicerae (Seeler) Hirooka, Rossman & P. Chaverri, Pleonectria rosellinii (Carestia) Hirooka, Rossman & P. Chaverri, Pleonectria rubicarpa (Cooke) Hirooka, Rossman & P. Chaverri, Pleonectria sinopica (Fr.: Fr.) Hirooka, Rossman & P. Chaverri, Pleonectria sphaerospora (Ellis & Everh) Hirooka, Rossman & P. Chaverri, Pleonectria virens (Harkn.) Hirooka, Rossman & P. Chaverri, Pleonectria zanthoxyli (Peck) Hirooka, Rossman & P. Chaverri.

Nectria flute canker is a disease of Pinus radiata stems caused by the pathogen Neonectria fuckeliana occurring in the southern parts of New Zealand. In Northern Hemisphere countries where N. fuckeliana is endemic, it is commonly found in Picea and Abies spp. Open wounds, dead attached branches and branch stubs have been identified as the primary infection courts. Although in New Zealand the development of Nectria flute canker disease is associated with pruned branch stubs, recent studies suggest that this is not the only possible entry method as the fungus has been found in trees prior to pruning. Three field trials were established to examine the potential infection mechanisms for N. fuckeliana in P. radiata in New Zealand; including stem wounds and branch stubs. The difference between inoculations into the stem and into branch wood was clear. Inoculation of deep stem wounds resulted in the greatest fluting with 76% of trees inoculated developing cankers. Inoculation directly into stubs resulted in only small stem depressions that occurred in 17% of cases and the fungus was largely contained within the branch trace. Tree response to inoculation with either ascospores or conidia of the Acremonium anamorph gave similar results in terms of canker development and fungal spread within the stem. Tree response to inoculation was highly variable however: in one study 6% of trees did not respond to inoculation at all, while 26% produced severe cankers regardless of inoculation method. A more thorough understanding of the infection mechanisms of N. fuckeliana will contribute to the development of better disease management protocols to prevent infection and disease development in future plantation stock.

Nectria flute canker is a fungal disease that results in cankers on Pinus radiata stems. The disease, caused by the pathogen Neonectria fuckeliana, is associated with commercial pruning operations that limit knot defect to a central core to promote growth of high-quality clearwood. Nectria flute canker is present in commercial plantation forests in parts of New Zealand and Chile. Symptoms of the disease, bark cracks, stem flattenings or depressions and flute cankers, were identified on 25 mature Pinus radiata trees. Trees were felled, and discs crosscut from within, and adjacent to, the disease symptoms. The discs were photographed, and three different methods were used to determine the extent of Nectria damage beyond the defect core into the valuable clearwood sheath. These methods included a visual assessment of photographs of internal damage and two quantitative methods, based on calculations of defect extent using image processing software. Overall, flute cankers were associated with the greatest degree of encased bark and severe decay pockets, and stem flattenings the least. None of the flattened stems had internal decay. The average damage to the clearwood sheath due to flute canker was 43%, whilst that for bark cracks and flattenings was 19 and 8%, respectively. Similar results were achieved using all three methods, and the preferred method would depend on the context of the study. Using a combination of all three methods provides a robust method for measuring the damage to internal wood quality by stem pathogens and other abiotic agents.

Airborne ascospores have been reported to be allergenic or plant pathogens, and their presence has traditionally been associated with rainfall events. The aim of the present study was to analyze the presence of airborne ascospores in relation to weather parameters in a town in SW Spain. A seven-day recording spore trap (Burkard) was used to sample the air over 2 years at 15 m above ground level on the terrace roof of the hospital in Merida (SW Spain). Fungal spores were identified and counted by means of two longitudinal scans over the slides under ×1000 microscopy. A correlation analysis was made of the daily meteorological data and the airborne ascospore concentrations, and t-tests were used to compare data between the 2 years. Nineteen ascospore types were defined, including one-cell ascospores (Chaetomium, Diatrype, Helvella, Xylaria), tow-cell ascospores (Diaporthe, Mycosphaerella, Nectria, Venturia), transversally septate ascospores (Melanomma, Leptosphaeria, Paraphaeosphaeria, Sporormiella, Massaria), transversally and longitudinally septate ascospores (Pleospora), and ascospores within asci (Sordaria). Leptosphaeria consisted of a group of four types described according to the number of cells, hyaline grade, wall thickness, and ornamentation, and other ascospores comprised one last additional type. The average airborne ascospore concentration was 153 ascospores/m³. One-third each of this total were from the Leptosphaeria group, with an average 54 ascospores/m³, and the two-cell ascospores or Venturia-like group (Diaporthe, Mycosphaerella, Nectria, Venturia) with 51 ascospores/m³ on average. In third position was Pleospora with 27 ascospores/m³ on average. The month with highest concentration was September, with 238 ascospores/m³, and the lowest March, with 56 ascospores/m³. By seasons, autumn had the highest concentrations, followed by winter, spring, and summer. The maximum daily concentration reached was 3,371 ascospores/m³. Daily rainfall was significantly correlated with the ascospore types Diatrype, Mycosphaerella, Nectria, two subtypes of Leptosphaeria, and Pleospora. Relative humidity was positively correlated with those ascospore types and also with Diaporthe and Paraphaeosphaeria, and negatively with Chaetomium and Melanomma. The concentration was higher on rainy days than on days without rain for Pleospora, Leptosphaeria (3 subtypes), Diatrype, Diaporthe, Nectria, Mycosphaerella, and Paraphaeosphaeria. The daily temperatures were in general correlated with the same types as the relative humidity, but with the opposite sign. For the monthly data, there were no statistically significant differences between the 2 years studied.

Neonectria is a cosmopolitan genus and it is, in part, defined by its link to the anamorph genus Cylindrocarpon. Neonectria has been divided into informal groups on the basis of combined morphology of anamorph and teleomorph. Previously, Cylindrocarpon was divided into four groups defined by presence or absence of microconidia and chlamydospores. Molecular phylogenetic analyses have indicated that Neonectria sensu stricto and Cylindrocarpon sensu stricto are phylogenetically congeneric. In addition, morphological and molecular data accumulated over several years have indicated that Neonectria sensu lato and Cylindrocarpon sensu lato do not form a monophyletic group and that the respective informal groups may represent distinct genera. In the present work, a multilocus analysis (act, ITS, LSU, rpb1, tef1, tub) was applied to representatives of the informal groups to determine their level of phylogenetic support as a first step towards taxonomic revision of Neonectria sensu lato. Results show five distinct highly supported clades that correspond to some extent with the informal Neonectria and Cylindrocarpon groups that are here recognised as genera: (1) N. coccinea-group and Cylindrocarpon groups 1 & 4 (Neonectria/Cylindrocarpon sensu stricto); (2) N. rugulosa-group (Rugonectria gen. nov.); (3) N. mammoidea/N. veuillotiana-groups and Cylindrocarpon group 2 (Thelonectria gen. nov.); (4) N. radicicola-group and Cylindrocarpon group 3 (Ilyonectria gen. nov.); and (5) anamorph genus Campylocarpon. Characteristics of the anamorphs and teleomorphs correlate with the five genera, three of which are newly described. New combinations are made for species where their classification is confirmed by phylogenetic data.Taxonomic novelties:Ilyonectria P. Chaverri & C. Salgado, gen. nov.; Ilyonectria coprosmae (Dingley) P. Chaverri & C. Salgado, comb. nov.; Ilyonectria liriodendri (Halleen et al.) P. Chaverri & C. Salgado, comb. nov.; Ilyonectria macrodydima (Halleen, Schroers & Crous) P. Chaverri & C. Salgado, comb. nov.; Ilyonectria radicicola (Gerlach & L. Nilsoon) P. Chaverri & C. Salgado, comb. nov.; Rugonectria P. Chaverri & Samuels, gen. nov.; Rugonectria castaneicola (W. Yamam. & Oyasu) Hirooka & P. Chaverri, comb. nov; Rugonectria neobalansae (Samuels) P. Chaverri & Samuels, comb. nov.; Rugonectria rugulosa (Pat. & Gaill.) Samuels, P. Chaverri & C. Salgado, comb. nov.; Thelonectria P. Chaverri & C. Salgado, gen. nov.; Thelonectria coronata (Penz. & Sacc.) P. Chaverri & C. Salgado, comb. nov.; Thelonectria discophora (Mont.) P. Chaverri & C. Salgado, comb. nov.; Thelonectria jungneri (Henn.) P. Chaverri & C. Salgado, comb. nov.; Thelonectria lucida (Höhnel) P. Chaverri & C. Salgado, comb. nov.; Thelonectria olida (Wollenw.) P. Chaverri & C. Salgado, comb. nov; Thelonectria trachosa (Samuels & Brayford) Samuels, P. Chaverri & C. Salgado, comb. nov.; Thelonectria veuillotiana (Sacc. & Roum.) P. Chaverri & C. Salgado, comb. nov.; Thelonectria viridispora (Samuels & Brayford) P. Chaverri, C. Salgado, & Samuels, comb. nov.; Thelonectria westlandica (Dingley) P. Chaverri & C. Salgado, comb. nov.

Myrmaeciella caraganae was recollected in and around Vienna, Austria and found to be morphologically different generically from the type species of Myrmaeciella, M. endoleuca. We redescribe M. caraganae in the new genus Stromatonectria. Phylogenetic analyses of LSU sequences place the genus in the Bionectriaceae, Hypocreales. S. caraganae occurs on branches of Caragana spp., Colutea arborescens and Laburnum anagyroides of the Fabaceae. It is characterized by spheroid perithecia partly or entirely immersed in a Hypocrea-like stroma, a Nectria-like centrum and bicellular hyaline ascospores. Conidia of S. caraganae are produced in compound pycnidia that are formed prior to or in association with perithecia. Sporodochia but no pycnidia are formed in culture. We discuss the genus Myrmaeciella and compare S. caraganae with species of the Nectriaceae, including Nectria balansae, N. eustromatica and N. paraguayensis.

The fungal plant pathogen Nectria haematococca MPVI produces a cytochrome P450 that is responsible for detoxifying the phytoalexin pisatin, produced as a defense mechanism by its host, garden pea. In this study, we demonstrate that this fungus also produces a specific ATP-binding cassette (ABC) transporter, NhABC1, that enhances its tolerance to pisatin. In addition, although both mechanisms individually contribute to the tolerance of pisatin and act as host-specific virulence factors, mutations in both genes render the fungus even more sensitive to pisatin and essentially nonpathogenic on pea. NhABC1 is rapidly induced after treatment with pisatin in vitro and during infection of pea plants. Furthermore, NhABC1 was able to confer tolerance to the phytoalexin rishitin, produced by potato. NhABC1 appears to be orthologous to GpABC1 of the potato pathogen Gibberella pulicaris and, along with MoABC1 from Magnaporthe oryzae, resides in a phylogenetically related clade enriched with ABC transorters involved in virulence. We propose that NhABC1 and the cytochrome P450 may function in a sequential manner in which the energy expense from pisatin efflux by NhABC1 releases the repression of the cytochrome P450, ultimately allowing pisatin tolerance by two mechanisms. These results demonstrate that a successful pathogen has evolved multiple mechanisms to overcome these plant antimicrobial compounds.

Nectria flute canker is a disease of Pinus radiata stems caused by Neonectria fuckeliana. The fungus is not known in the North Island, leading to concern that it may be transported north via asymptomatically colonised nursery stock. Wounded P. radiata seedlings and cuttings were exposed to natural inoculum from an adjacent infected forest, and half were artificially inoculated. No disease symptoms were recorded throughout the trial. Neonectria fuckeliana was re-isolated from 24% and 7% of inoculated seedlings and cuttings respectively 3 months after inoculation, and at 12 months from 3.5% of the remaining inoculated seedlings only. Neonectria fuckeliana was not isolated from uninoculated seedlings or cuttings exposed to natural inoculum. These results indicate that nursery plants have little capability of retaining N. fuckeliana within the tissue even when directly inoculated. Nursery stock is therefore unlikely to be a pathway for transport of the fungus.

A series of small-scale controlled inoculation experiments has been conducted during 2005–2009 to determine whether temperature and controlled atmosphere (CA) storage conditions affect significantly the incidence of Botrytis cinerea and Neonectria galligena rots of apples and to assess whether CA regimes can be ‘fine-tuned’ to suppress fungal rotting. The incidence of B. cinerea and N. galligena rots on apple was reduced consistently by storage in lower temperatures (1.5–2°C). In no case was the disease incidence significantly higher than that under air storage conditions. However, the effect of CA conditions on rot development varied greatly from year to year so that overall there were no significant effects of CA conditions on the incidence of rot during storage till the following April. Thus, we can neither fine-tune CA conditions to reduce rot development nor exploit the advantages of lower storage temperatures for Cox and Bramley for this purpose due to inevitable development of low temperature breakdown. Further research is needed to study the effects of ethylene control technologies and modulated storage temperatures on rot development.

A comprehensive phylogenetic reassessment of the ascomycete genus Cosmospora (Hypocreales, Nectriaceae) is undertaken using fresh isolates and historical strains, sequences of two protein encoding genes, the second largest subunit of RNA polymerase II (rpb2), and a new phylogenetic marker, the larger subunit of ATP citrate lyase (acl1). The result is an extensive revision of taxonomic concepts, typification, and nomenclatural details of many anamorph- and teleomorph-typified genera of the Nectriaceae, most notably Cosmospora and Fusarium. The combined phylogenetic analysis shows that the present concept of Fusarium is not monophyletic and that the genus divides into two large groups, one basal in the family, the other terminal, separated by a large group of species classified in genera such as Calonectria, Neonectria, and Volutella. All accepted genera received high statistical support in the phylogenetic analyses. Preliminary polythetic morphological descriptions are presented for each genus, providing details of perithecia, micro- and/or macro-conidial synanamorphs, cultural characters, and ecological traits. Eight species are included in our restricted concept of Cosmospora, two of which have previously documented teleomorphs and all of which have Acremonium-like microconidial anamorphs. A key is provided to the three anamorphic species recognised in Atractium, which is removed from synonymy with Fusarium and epitypified for two macroconidial synnematous species and one sporodochial species associated with waterlogged wood. Dialonectria is recognised as distinct from Cosmospora and two species with teleomorph, macroconidia and microconidia are accepted, including the new species D. ullevolea. Seven species, one with a known teleomorph, are classified in Fusicolla, formerly considered a synonym of Fusarium including members of the F. aquaeductuum and F. merismoides species complex, with several former varieties raised to species rank. Originally a section of Nectria, Macroconia is raised to generic rank for five species, all producing a teleomorph and macroconidial anamorph. A new species of the Verticillium-like anamorphic genus Mariannaea is described as M. samuelsii. Microcera is recognised as distinct from Fusarium and a key is included for four macroconidial species, that are usually parasites of scale insects, two of them with teleomorphs. The four accepted species of Stylonectria each produce a teleomorph and micro- and macroconidial synanamorphs. The Volutella species sampled fall into three clades. Pseudonectria is accepted for a perithecial and sporodochial species that occurs on Buxus. Volutella s. str. also includes perithecial and/or sporodochial species and is revised to include a synnematous species formerly included in Stilbella. The third Volutella-like clade remains unnamed. All fungi in this paper are named using a single name system that gives priority to the oldest generic names and species epithets, irrespective of whether they are originally based on anamorph or teleomorph structures. The rationale behind this is discussed.Taxonomic novelties: New genus:Macroconia (Wollenw.) Gräfenhan, Seifert & Schroers. New species:Dialonectria ullevolea Seifert & Gräfenhan, Fusicolla violacea Gräfenhan & Seifert, Mariannaea samuelsiiSeifert & Bissett, Microcera rubra Gräfenhan & Seifert. New combinations:Atractium holubovae (Seifert, S.J. Stanley & K.D. Hyde) Seifert, Atractium crassum (Wollenw.) Seifert & Gräfenhan, Cosmospora arxii (W. Gams) Gräfenhan & Schroers, Cosmospora berkeleyana (P. Karst.) Gräfenhan, Seifert & Schroers, Cosmospora butyri (J.F.H, Beyma) Gräfenhan, Seifert & Schroers, Cosmospora cymosa (W. Gams) Gräfenhan & Seifert, Cosmospora khandalensis (Thirum. & Sukapure) Gräfenhan & Seifert, Cosmospora lavitskiae (Zhdanova) Gräfenhan & Seifert, Cosmospora viridescens (C. Booth) Gräfenhan & Seifert, Fusicolla acetilerea(Tubaki, C. Booth & T. Harada) Gräfenhan & Seifert, Fusicolla aquaeductuum (Radlk. & Rabenh.) Gräfenhan, Seifert & Schroers, Fusicolla epistroma (Höhn.) Gräfenhan & Seifert, Fusicolla matuoi (Hosoya & Tubaki) Gräfenhan & Seifert, Fusicolla merismoides (Corda) Gräfenhan, Seifert & Schroers, Macroconia cupularis (J. Luo & W.Y. Zhuang) Gräfenhan & Seifert, Macroconia gigas (J. Luo & W.Y. Zhuang) Gräfenhan & Seifert, Macroconia leptosphaeriae (Niessl) Gräfenhan & Schroers, Macroconia papilionacearum (Seaver) Gräfenhan & Seifert, Macroconia sphaeriae (Fuckel) Gräfenhan & Schroers, Microcera diploa (Berk. & M.A. Curtis) Gräfenhan & Seifert, Microcera larvarum (Fuckel) Gräfenhan, Seifert & Schroers, Pseudonectria buxi (DC.) Seifert, Gräfenhan & Schroers, Stylonectria purtonii (Grev.) Gräfenhan, Stylonectria wegeliniana (Rehm) Gräfenhan, Voglmayr & Jaklitsch, Volutella citrinella (Ellis & Everh.) Seifert, Volutella consors (Ellis & Everh.) Seifert, Gräfenhan & Schroers. New name:Stylonectria carpini Gräfenhan.

A new species with remarkable morphology, Nectria eustromatica, is described, based on morphology of the teleomorph and anamorph, ecology and molecular phylogenetic analyses. Nectria eustromatica is characterized by sphaeroid perithecia immersed in pseudoparenchymatous stromata formed singly or collectively on a subiculum. Despite its deviating teleomorph morphology, it is placed within Nectria sensu stricto in phylogenetic analyses of a combined dataset of LSU, ITS, rpb2 and tef1 sequences with high internal support. Nectria eustromatica has been collected specifically on Hippocrepis (Coronilla) emerus in southern Europe. The anamorph of N. eustromatica shares morphological traits with the genera Stilbella and Tubercularia but produces non-phialidic macroconidia in addition to phialoconidia.

Lung cancer is a common disease that is associated with poor prognosis. Fungal immunomodulatory protein from Nectria haematococca (FIP-nha) has potential as a lung cancer therapeutic, as such, illuminating its anti-tumor mechanism is expected to facilitate novel treatment options. Here, we showed that FIP-nha affects lung adenocarcinoma growth ex vivo and in vivo. Comparative quantitative proteomics showed that FIP-nha negatively regulates PI3K/Akt signaling and induces cell cycle arrest, autophagy, and apoptosis. We further demonstrated that FIP-nha suppresses Akt phosphorylation, leading to upregulation of p21 and p27 and downregulation of cyclin B1, cyclin D1, CDK2, and CDK4 expression, ultimately resulting in G1/S and G2/M cell cycle arrest. Meanwhile, FIP-nha-induced PI3K/Akt downregulation promotes A549 apoptosis by increasing the expression ratio of Bax/Bcl-2 and c-PARP and autophagy by decreasing the phosphorylation of mTOR. Thus, we comprehensively revealed the anti-tumor mechanism of FIP-nha, which inhibits tumor growth by modulating PI3K/Akt-regulated cell cycle arrest, autophagy, and apoptosis, and provided the basis for further application of fungal immunomodulatory proteins, especially FIP-nha.

This study investigates the diversity of fungal endophytes associated with several varieties of grapevines with differing modes of cultivation in the Madrid region (central Spain). Our goal is to screen for and identify new fungal biocontrol agents against vine diseases, especially those associated with young plants produced in nurseries. A total of 500 fungal strains representing 68 taxa from six locations were isolated and characterised. Differences regarding cultivar and plant part processed were analysed in terms of composition and relative abundance of species. Some of the more frequently isolated strains represented were Acremonium, Alternaria, Aureobasidium, Botryotinia, Cladosporium, Epicoccum, Fusarium, Gibberella, Nectria, Penicillium, Phoma and Trichoderma species. Botryosphaeria species and Phomopsis viticola were also frequently isolated and may be vine pathogens. Several species of Acremonium, Phoma (P. glomerata) and Chaetomium showed promising antagonistic activity at the laboratory scale.

We used 454 sequencing of the internal transcribed spacer region to characterize fungal communities in tallgrass prairie soils subdivided into strata 0-10, 10-20, 30-40 and 50-60 cm deep. The dataset included more than 14000 fungal sequences distributed across Basidiomycota, Ascomycota, basal fungal lineages and Glomeromycota in order of decreasing frequency. As expected the community richness and diversity estimators tended to decrease with increasing depth. Although species richness was significantly reduced for samples from the deeper profiles, even the deepest stratum sampled contained richness of more than a third of that in the topmost stratum. More importantly, nonparametric multidimensional scaling (NMS) ordination analyses indicated that the fungal communities differed across vertical profiles, although only the topmost and deepest strata were significantly different when the NMS axis scores were compared by ANOVA. These results emphasize the importance of considering the fungal communities across the vertical strata because the deeper soil horizons might maintain a distinct community composition and thus contribute greatly to overall richness. The majority of operational taxonomic units (OTUs) declined in frequency with increasing depth, although a linear regression analysis indicated that some increased with increasing depth. The OTUs and BLAST-assigned taxa that showed increasing frequencies were mainly unculturable fungi, but some showed likely affinities to families Nectriaceae and Venturiaceae or to genus Pachnocybe. Although the ecological roles of the fungi in the deeper strata remain uncertain, we hypothesize that the fungi with preferences for deeper soil have adequate access to substrates and possess environmental tolerances that enable their persistence in those environments.

The muck soils of the north shore of Lake Apopka, near Orlando, Florida, USA are high in organic matter, inorganic nutrients, and water content. Ideally suited for agriculture, these soils have been exposed to a wide variety of agrochemicals. Some of the more recalcitrant organochlorine pesticides, such as DDT and its degradation products DDD and DDE, have persisted in the soil for over 30 years. Using the extracellular enzymes from wood rot fungi, it was demonstrated that it is possible to substantially reduce the amount of the o,p′ and p,p′ isomers of DDT, DDD, and DDE in this soil by more than 60% in 3 weeks. A fungal species with 99% DNA homology to Nectria mariannaeae was isolated from this muck soil and identified by nucleotide sequencing. When grown under nitrogen-limited conditions, this Nectria sp. has been shown to be comparable to Phanerochaete chrysosporium (ATCC 24725) in producing extracellular factors (or agents) that are capable of degrading these recalcitrant chlorinated chemicals.

Nectria is a wood-inhabiting genus of fungi commonly found in temperate regions of the northern hemisphere, where they are associated with dieback and canker diseases of numerous woody plant hosts. In this study, fungal isolates were collected from 2012 to 2017 from diseased branches or twigs of 19 different host species in China and identified. Their morphological characteristics and multi-locus phylogeny (act, ITS, LSU, rpb2, tef1, and tub2) indicated four distinct lineages with high statistical support, corresponding to four species of Nectria: N. balansae, N. dematiosa, N. pseudotrichia, and N. ulmicola sp. nov. The last is characterized by yellow to orange, or sometimes dark brown, sporodochial conidiomata, which are sessile, and by unbranched or bifurcate conidiophores. This study has clarified the taxonomy of Nectria species associated with canker and dieback symptoms in China.

Neonectria galligena can cause European canker of apple as well as fruit rot. Healthy unwounded fruits on potted trees of cvs Cox, Bramley and Gala were inoculated with conidia of N. galligena to investigate the effects of wetness duration and fruit maturity on rot development. Overall, the incidence of fruit rot was influenced more by fruit maturity at the time of inoculation than by duration of wetness (6–48 h). Young fruit were most susceptible to infection, with 50% of fruit infected when inoculated up to 4 weeks after full bloom. The susceptibility decreased initially until c. 2 months after full bloom and then increased gradually until harvest. Almost all preharvest symptoms (eye rot) developed only on the fruit inoculated up to 4 weeks after full bloom. All other rots were observed after six-month postharvest storage under controlled atmospheric conditions. However, the relative proportion of preharvest eye rots and postharvest storage rots varied greatly among three years. The effect of wetness duration was only significant for fruit inoculated in their early stages of development but not for those inoculated near harvest. Regression models were developed to describe the observed effects of fruit maturity and wetness on the incidence of total nectria rots.

Zusammenfassung Die Laubholz-Lachniden unterhalten sehr enge Trophobiosebeziehungen zu Ameisen. Ohne die Ameisen konnen sie auf die Dauer nicht existieren. Die Rote Waldameise Formica rufa ist in der Lage, in der Nahe ihrer Nester fast alle Rauber wirksam von den Lausen abzuwehren. Die wirksamsten Feinde sind Vogel, die wahrend des Winters regelmasig den grosten Teil der Schizodryobius-Gelege verzehren. L. roboris konnten in Deutschland noch keine Schaden nachgewiesen werden, wahrend er in Ungarn als Erreger des Eichenkropfes aufgetreten sein soll. S. pallipes und S. longirostris bewirken durch ihre Saugtatigkeit eine Entartung des Kambiums ihrer Nahrpflanzen. An Buche fuhrt das zu Risbildungen und im Zusammenwirken mit dem Buchenkrebspilz Nectria ditissima zum Absterben von Zweigen und jungeren Pflanzen. An Eiche entstehen Rindenverfarbungen, Ris- und Kropfbildungen und eine Reihe weiterer Schaden, die ebenfalls das Absterben von Zweigen oder Jungpflanzen nach sich ziehen konnen. Summary The Lachnids of deciduous trees maintain very close trophibiotic relationships with ants and cannot exist for long without these insects. Near their nests, red wood ants, Formica rufa, can effectively protect the aphids from almost all predators. The most effective enemies are birds which regularly consume the greater part of the Schizodryobius egg masses in the winter. Up to now, it has not been possible to prove any damage by L. roboris in Germany, whereas this appears to be the cause of oack canker (Eichenkropf) in Hungary. By their feeding activities, S. pallipes and S. longirostris cause a degeneration of the cambium of their host trees. On beech, this leads to crack formation and, in conjunction with the beech canker fungus, Nectria ditissima, to the death of twigs and young trees. On oak, bark discoloration, crack and canker (Kropf) formation and a number of further symptoms occur, which can also cause the death of twigs and young trees.

Plant defense involves a complex array of biochemical interactions, many of which occur in the extracellular environment. The apical 1- to 2-mm root tip housing apical and root cap meristems is resistant to infection by most pathogens, so growth and gravity sensing often proceed normally even when other sites on the root are invaded. The mechanism of this resistance is unknown but appears to involve a mucilaginous matrix or “slime” composed of proteins, polysaccharides, and detached living cells called “border cells.” Here, we report that extracellular DNA (exDNA) is a component of root cap slime and that exDNA degradation during inoculation by a fungal pathogen results in loss of root tip resistance to infection. Most root tips (>95%) escape infection even when immersed in inoculum from the root-rotting pathogen Nectria haematococca. By contrast, 100% of inoculated root tips treated with DNase I developed necrosis. Treatment with BAL31, an exonuclease that digests DNA more slowly than DNase I, also resulted in increased root tip infection, but the onset of infection was delayed. Control root tips or fungal spores treated with nuclease alone exhibited normal morphology and growth. Pea (Pisum sativum) root tips incubated with [32P]dCTP during a 1-h period when no cell death occurs yielded root cap slime containing 32P-labeled exDNA. Our results suggest that exDNA is a previously unrecognized component of plant defense, an observation that is in accordance with the recent discovery that exDNA from white blood cells plays a key role in the vertebrate immune response against microbial pathogens.

Aim: The aim of this study was to develop molecular assays for investigating the presence and diversity of pathogenicity genes from the pea footrot pathogen Nectria haematococca (anamorph Fusarium solani f.sp. pisi) in soils. Methods and Results: Polymerase chain reaction (PCR) assays were developed to amplify four N. haematococca pathogenicity genes (PDA, PEP1, PEP3 and PEP5) from isolates and soil-DNA from five agricultural fields with a prior footrot history. A collection of 15 fungi isolated on medium selective for Fusarium spp. exhibited variation in their virulence to peas as assessed via a disease index (DI: 0-5; no virulence to the highest virulence). PCR analyses showed that three isolates in which all four pathogenicity genes were detected resulted in the highest DI (>3·88). All four pathogenicity genes were detected in soil-DNA obtained from all five fields with a footrot disease history, but were not amplified from soils, which had no footrot history. Denaturing gradient gel electrophoresis and/or sequence analysis revealed diversity amongst the pathogenicity genes. Conclusion: The PCR assays developed herein enable the specific detection of pathogenic N. haematococca in soils without recourse to culture. Significance and Impact of the Study: Molecular assays that specifically target pathogenicity genes have the capacity to assess the presence of the footrot-causing pathogen in agricultural soils.

Stands of eastern hemlock [(Tsuga canadensis (L.) Carriere] in the north-eastern United States are in decline, in part from the attack of elongate hemlock scale, Fiorinia externa Ferris (Hemiptera: Diaspididae). From 2001 to the present, a natural epizootic has been observed in populations of F. externa. Initially discovered at the Mianus River Gorge Preserve in Bedford, New York, the epizootic has also been detected in Pennsylvania, New Jersey and Connecticut. Understanding and assessing the identity of the pathogenic micro-organisms responsible for this natural mortality is crucial for developing biological controls for this pest. We have isolated and taxonomically and genetically identified entomopathogens, phytopathogens and endophytic fungi associated with F. externa. Isolates of the following were obtained: Colletotrichum sp., Lecanicillium lecanii, Beauveria bassiana, Metarhiziopsis microspora, Myriangium sp., Mycosphaerella sp. anamorph, Nectria sp., Botrytis sp., Phialophora sp. and Fusarium sp.

Ten species of lichenicolous fungi on Collemataceae in the Iberian Peninsula are recorded. A new species, Pronectria pilosa, on Collema furfuraceum is proposed and three new records from the Iberian Peninsula are indicated, Endococcus pseudocarpus, Nectria brutia and Toninia leptogii. The known distribution of several treated species is enlarged notably.

The new genus Cyanonectria is proposed for Nectria cyanostoma (≡ Cyanonectria cyanostoma comb. nov.). This genus is characterized by Nectria-like, red perithecia that have a bluish-purple papilla and a Fusarium anamorph. DNA sequences (large subunit and internal transcribed spacers of the nuclear rDNA) indicate that C. cyanostoma is not closely related to Nectria sensu stricto. In addition, the phylogenetic data also show that the closest relatives for Cyanonectria also have Fusarium anamorphs.

Fusarium stem canker of greenhouse eggplant, incited by Fusarium solani (teleomorph Nectria haematacocca), caused extensive losses in yield in a commercial operation in Leamington, Ontario, in 2005. Identification of the disease was based on host symptoms, reisolation of the pathogen, morphological and cultural characteristics of the fungus, and the fulfillment of Koch's postulates. Control measures are discussed. This is the first report of this disease on greenhouse eggplant in Canada. Key words: Fusarium solani, Nectria haematacocca, stem canker, greenhouse, eggplant. Le Fusarium d'aubergine de serre, incite par Fusarium solani (teleomorph Nectria haematacocca), a cause des pertes considerables de rendement en 2005 dans le milieu de serre commercial de la region de Leamington (Ontario, Canada). L'identification a ete basee sur des symptomes de la hote, reisolation du microbe pathogene, des caracteristiques morphologiques et culturelles et aussi en se basant sur les postulats de Koch. Des mesures de con...

Fungi and bacteria on the external surfaces of four gorse-associated insect species: gorse seed weevil Apion ulicis Forster (Coleoptera: Apionidae), light brown apple moth Epiphyas postvittana Walker (Lepidoptera: Tortricidae), gorse pod moth Cydia ulicetana Denis and Schiffermuller (Lepidoptera: Tortricidae) and gorse thrips Sericothrips staphylinus Haliday (Thysanoptera: Thripidae), were recov- ered by washing and plating techniques. The isolates were identified by morphology and polymerase chain reaction (PCR) restriction fragment length polymorphism and sequencing of internally tran- scribed spacer (ITS) and 16S rDNA. A culture-independent technique (direct PCR) was also used to assess fungal diversity by direct amplification of ITS sequences from the washings of the insects. All insect species carried Alternaria, Cladosporium, Corallomycetella, Penicillium, Phoma, Pseudozyma spp. and entomopathogens. Ninety-four per cent of the 178 cloned amplicons had ITS sequence similarity to Nectria mauritiicola (syn. Corallomycetella repens) .E. postvittana carried the largest fungal spores (spore mean surface area of 126 mm 2 ) and the most fungal colony forming units per insect. Methylo- bacterium aquaticum and Pseudomonas lutea were isolated from all four insect species. P. fluorescens was the most abundant bacterium on the lepidopteran insects. This study presents the diversity of microbial taxa on insect exoskeletons, and provides the basis for developing a novel mycoherbicide delivery strategy for biological control of gorse using insects as vectors of a plant pathogen.

BACKGROUND: The novel natural product cinnacidin was isolated from a fungal fermentation extract of Nectria sp. DA060097. The compound was found to contain a cyclopentalenone ring system with an isoleucine subunit linked through an amide bond. Initial biological characterization of cinnacidin suggested promising herbicidal activity. RESULTS: Two synthetic analogs, (2S,3S)-2-[(3RS,3aSR,6aRS)-3-methoxy-4-oxo-3,3a,4,5,6,6a-hexahydropentalen-1-ylcarbamoyl]-3-methylvaleric acid and benzyl (2S,3S)-2-[(3RS,3aSR,6aRS)-3-methoxy-4-oxo-3,3a,4, 5,6,6a-hexahydropentalen-1-ylcarbamoyl]-3-methylvalerate, were prepared for further characterization, and their herbicidal activities were compared with that of cinnacidin. CONCLUSIONS: The synthetic compounds were highly phytotoxic on a range of weeds. Based on the symptoms in treated plants, the mode of action of these compounds is suggested to be similar to that of coronatine and jasmonic acid. Coronatine was more active against warm-season grasses, while the cinnacidin benzyl ester analog was more effective on cool-season grasses. In a seedling growth bioassay conducted on bentgrass, the cinnacidin analog was equivalent in activity to coronatine. Copyright © 2008 Society of Chemical Industry

Aims: The aim of the study was to isolate and characterize the endophytic fungi from the rhizomes of the Chinese traditional medicinal plant Dioscorea zingiberensis and to detect their antibacterial activities. Methods and Results: After strict sterile sample preparation, nine fungal endophytes were isolated from rhizomes of the Chinese traditional medicinal plant D. zingiberensis. The endophytes were classified by morphological traits and internal transcribed spacer (ITS) rRNA gene sequence analysis. Their ITS rDNA sequences were 99–100% identical to Nectria, Fusarium, Rhizopycnis, Acremonium and Penicillium spp. respectively. Of these, the most frequent genera were Fusarium and Nectria. One isolate, Dzf7, was unclassified on the basis of its low sequence similarity. The next closest species was Alternaria longissima (c. 92·4% sequence similarity). Endophyte isolate Dzf5 showed the closest sequence similarity (c. 99·5%) to an uncultured soil fungus (DQ420800) obtained from Cedar Creek, USA. Bioassays using a modified broth dilution test were used to detect the antibacterial activity of n-butanol extracts of both mycelia and culture filtrates of D. zingiberensis showed biological activity against Bacillus subtilis, Staphylococcus haemolyticus, Escherichia coli and Xanthomonas vesicatoria. Minimal inhibitory concentration (MIC) values of the extracts were between 31·25 μg ml−1 and 125 μg ml−1. Conclusions: Endophytic fungus Dzf2 (c. 99·8% sequence similarity to Fusarium redolens) isolated from D. zingiberensis rhizome showed the most potent antibacterial activities. Significance and Impact of the Study: Endophytic fungi isolated from D. zingiberensis may be used as potential producers of antibacterial natural products.

Fusarium species cause dry rot or seedpiece decay of potato tubers in storage and in the field. When potato storages were sampled in the Columbia Basin of Oregon and Washington,F. sambucinum, F. oxysporum, andF. solani were the predominantFusarium species recovered from potato tubers with dry rot. OtherFusarium species recovered includedF. avenaceum, F. culmorum, F. equiseti, F. proliferatum, andF. sporotrichioides. The majority ofF. sambucinum isolates tested were resistant to thiabendazole at 5, 10, and 25 mg liter-1 in amended potato dextrose agar (PDA), whereas resistance was less frequent inF. oxysporum andF. solani isolates. When isolates ofF. sambucinum, F. solani, andF. oxysporum were grown on PDA with no thiabendazole, the average colony diameter of isolates with resistance to thiabendazole was greater than the average colony diameter of isolates sensitive to thiabendazole for each respective species. When isolates ofF. sambucinum were placed on tuber slices, lesions were apparent by four days after inoculation, and lesion diameters were generally greater than those associated withF. solani orF. oxysporum. At eight days after inoculation onto tuber slices, the average lesion diameter ofF. sambucinum, F. solani, orF. oxysporum isolates with resistance to thiabendazole was greater than the average lesion diameter of isolates sensitive to thiabendazole for each respective species. Thiabendazole-resistance was also detected inF. avenaceum, F. culmorum, F. equiseti, andF. sporotrichioides.

Fruit tree canker (Neonectria ditissima syn. Nectria galligena Bres.) is an important fungal disease in apple (Malus × domestica Borkh.). Leaf fall is the predominant infection period for this disease. However, in The Netherlands and Belgium, no historical data on leaf fall of apple are available. In practice, collecting leaf fall data is a problem. An innovative approach was chosen to collect leaf fall data and introduce the Nectria model, according to Latorre, to fruit tree growers at the same time. An Android 2.2 Nectria application was developed. During leaf fall, growers received advice twice a week and used the smartphone application to send data and pictures, directly from the field. The general response of the first fruit tree growers who used the smartphone was very positive.