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Sample records for ascomycete podospora anserina

  1. The genome sequence of the model ascomycete fungus Podospora anserina

    PubMed Central

    Espagne, Eric; Lespinet, Olivier; Malagnac, Fabienne; Da Silva, Corinne; Jaillon, Olivier; Porcel, Betina M; Couloux, Arnaud; Aury, Jean-Marc; Ségurens, Béatrice; Poulain, Julie; Anthouard, Véronique; Grossetete, Sandrine; Khalili, Hamid; Coppin, Evelyne; Déquard-Chablat, Michelle; Picard, Marguerite; Contamine, Véronique; Arnaise, Sylvie; Bourdais, Anne; Berteaux-Lecellier, Véronique; Gautheret, Daniel; de Vries, Ronald P; Battaglia, Evy; Coutinho, Pedro M; Danchin, Etienne GJ; Henrissat, Bernard; Khoury, Riyad EL; Sainsard-Chanet, Annie; Boivin, Antoine; Pinan-Lucarré, Bérangère; Sellem, Carole H; Debuchy, Robert; Wincker, Patrick; Weissenbach, Jean; Silar, Philippe

    2008-01-01

    Background The dung-inhabiting ascomycete fungus Podospora anserina is a model used to study various aspects of eukaryotic and fungal biology, such as ageing, prions and sexual development. Results We present a 10X draft sequence of P. anserina genome, linked to the sequences of a large expressed sequence tag collection. Similar to higher eukaryotes, the P. anserina transcription/splicing machinery generates numerous non-conventional transcripts. Comparison of the P. anserina genome and orthologous gene set with the one of its close relatives, Neurospora crassa, shows that synteny is poorly conserved, the main result of evolution being gene shuffling in the same chromosome. The P. anserina genome contains fewer repeated sequences and has evolved new genes by duplication since its separation from N. crassa, despite the presence of the repeat induced point mutation mechanism that mutates duplicated sequences. We also provide evidence that frequent gene loss took place in the lineages leading to P. anserina and N. crassa. P. anserina contains a large and highly specialized set of genes involved in utilization of natural carbon sources commonly found in its natural biotope. It includes genes potentially involved in lignin degradation and efficient cellulose breakdown. Conclusion The features of the P. anserina genome indicate a highly dynamic evolution since the divergence of P. anserina and N. crassa, leading to the ability of the former to use specific complex carbon sources that match its needs in its natural biotope. PMID:18460219

  2. Integrative transformation of the ascomycete Podospora anserina: identification of the mating-type locus on chromosome VII of electrophoretically separated chromosomes.

    PubMed

    Osiewacz, H D; Skaletz, A; Esser, K

    1991-04-01

    Protoplasts of wild-type strain s and a long-lived extrachromosomal mutant (AL2) of the ascomycete Podospora anserina were transformed using a plasmid (pAN7-1) which contains the hygromycin B phosphotransferase gene (hph) of Escherichia coli under the control of Aspergillus nidulans regulatory sequences. After optimizing the transformation procedure, transformation efficiencies of 15-21 transformants/micrograms plasmid DNA were obtained. Using a second selectable vector (pBT3), which contains the beta-tubuline gene of a benomyl-resistant Neurospora crassa mutant, the co-transformation rate was determined. Southern blot hybridization experiments revealed that the transforming plasmid became integrated into the genome of the recipient either as a single copy or as multiple copies. In addition, the data from molecular as well as from classical genetic analyses indicated that in independent transformants vector integration occurred at different positions. The mitotic and meiotic stability of transformants proved to be dependent on the number of integrated plasmid copies. Genetic analyses revealed a transformant in which the integrated vector is closely linked to the mating-type locus. Fractionation of whole chromosomes by pulsed field gel electrophoresis and subsequent hybridization of the immobilized DNAs against radiolabelled vector sequences indicated the largest of seven chromosomes as the chromosome containing the integrated vector and thus the mating-type locus. PMID:1367277

  3. Heterologous production of cellobiose dehydrogenases from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina and their effect on saccharification of wheat straw.

    PubMed

    Turbe-Doan, Annick; Arfi, Yonathan; Record, Eric; Estrada-Alvarado, Isabel; Levasseur, Anthony

    2013-06-01

    Cellobiose dehydrogenases (CDHs) are extracellular glycosylated haemoflavoenzymes produced by many different wood-degrading and phytopathogenic fungi. Putative cellobiose dehydrogenase genes are recurrently discovered by genome sequencing projects in various phylogenetically distinct fungi. The genomes from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina were screened for candidate cdh genes, and one and three putative gene models were evidenced, respectively. Two putative cdh genes were selected and successfully expressed for the first time in Aspergillus niger. CDH activity was measured for both constructions (CDHcc and CDHpa), and both recombinant CDHs were purified to homogeneity and subsequently characterised. Kinetic constants were determined for several carbohydrates including β-1,4-linked di- and oligosaccharides. Optimal temperature and pH were 60 °C and 5 for CDHcc and 65-70 °C and 6 for CDHpa. Both CDHs showed a broad range of pH stability between 4 and 8. The effect of both CDHs on saccharification of micronized wheat straw by an industrial Trichoderma reesei secretome was determined. The addition of each CDH systematically decreased the release of total reducing sugars, but to different extents and according to the CDH concentration. Analytical methods were carried out to quantify the release of glucose, xylose and gluconic acid. An increase of glucose and xylose was measured at a low CDHcc concentration. At moderated and high CDHcc and CDHpa concentrations, glucose was severely reduced with a concomitant increase of gluconic acid. In conclusion, these results give new insights into the physical and chemical parameters and diversity of basidiomycetous and ascomycetous CDHs. These findings also demonstrated that CDH drastically influenced the saccharification on a natural substrate, and thus, CDH origin, concentration and potential enzymatic partners should be carefully considered in future artificial secretomes for biofuel applications. PMID:22940800

  4. Systematic Deletion of Homeobox Genes in Podospora anserina Uncovers Their Roles in Shaping the Fruiting Body

    PubMed Central

    Coppin, Evelyne; Berteaux-Lecellier, Vronique; Bidard, Frdrique; Brun, Sylvain; Ruprich-Robert, Gwenal; Espagne, Eric; At-Benkhali, Jinane; Goarin, Anne; Nesseir, Audrey; Planamente, Sara; Debuchy, Robert; Silar, Philippe

    2012-01-01

    Higher fungi, which comprise ascomycetes and basidiomycetes, play major roles in the biosphere. Their evolutionary success may be due to the extended dikaryotic stage of their life cycle, which is the basis for their scientific name: the Dikarya. Dikaryosis is maintained by similar structures, the clamp in basidiomycetes and the crozier in ascomycetes. Homeodomain transcription factors are required for clamp formation in all basidiomycetes studied. We identified all the homeobox genes in the filamentous ascomycete fungus Podospora anserina and constructed deletion mutants for each of these genes and for a number of gene combinations. Croziers developed normally in these mutants, including those with up to six deleted homeogenes. However, some mutants had defects in maturation of the fruiting body, an effect that could be rescued by providing wild-type maternal hyphae. Analysis of mutants deficient in multiple homeogenes revealed interactions between the genes, suggesting that they operate as a complex network. Similar to their role in animals and plants, homeodomain transcription factors in ascomycetes are involved in shaping multicellular structures. PMID:22662159

  5. The Transcriptional Response to Nonself in the Fungus Podospora anserina

    PubMed Central

    Bidard, Frdrique; Clav, Corinne; Saupe, Sven J.

    2013-01-01

    In fungi, heterokaryon incompatibility is a nonself recognition process occurring when filaments of different isolates of the same species fuse. Compatibility is controlled by so-called het loci and fusion of strains of unlike het genotype triggers a complex incompatibility reaction that leads to the death of the fusion cell. Herein, we analyze the transcriptional changes during the incompatibility reaction in Podospora anserina. The incompatibility response was found to be associated with a massive transcriptional reprogramming: 2231 genes were up-regulated by a factor 2 or more during incompatibility. In turn, 2441 genes were down-regulated. HET, NACHT, and HeLo domains previously found to be involved in the control of heterokaryon incompatibility were enriched in the up-regulated gene set. In addition, incompatibility was characterized by an up-regulation of proteolytic and other hydrolytic activities, of secondary metabolism clusters and toxins and effector-like proteins. The up-regulated set was found to be enriched for proteins lacking orthologs in other species and chromosomal distribution of the up-regulated genes was uneven with up-regulated genes residing preferentially in genomic islands and on chromosomes IV and V. There was a significant overlap between regulated genes during incompatibility in P. anserina and Neurospora crassa, indicating similarities in the incompatibility responses in these two species. Globally, this study illustrates that the expression changes occurring during cell fusion incompatibility in P. anserina are in several aspects reminiscent of those described in host-pathogen or symbiotic interactions in other fungal species. PMID:23589521

  6. Modulation of the glyoxalase system in the aging model Podospora anserina: effects on growth and lifespan

    PubMed Central

    Scheckhuber, Christian Q.; Mack, Sandra J.; Strobel, Ingmar; Ricciardi, Filomena; Gispert, Suzana; Osiewacz, Heinz D.

    2010-01-01

    The eukaryotic glyoxalase system consists of two enzymatic components, glyoxalase I (lactoylglutathione lyase) and glyoxalase II (hydroxyacylglutathione hydrolase). These enzymes are dedicated to the removal of toxic α-oxoaldehydes like methylglyoxal (MG). MG is formed as a by-product of glycolysis and MG toxicity results from its damaging capability leading to modifications of proteins, lipids and nucleic acids. An efficient removal of MG appears to be essential to ensure cellular functionality and viability. Here we study the effects of the genetic modulation of genes encoding the components of the glyoxalase system in the filamentous ascomycete and aging model Podospora anserina. Overexpression of PaGlo1 leads to a lifespan reduction on glucose rich medium, probably due to depletion of reduced glutathione. Deletion of PaGlo1 leads to hypersensitivity against MG added to the growth medium. A beneficial effect on lifespan is observed when both PaGlo1 and PaGlo2 are overexpressed and the corresponding strains are grown on media containing increased glucose concentrations. Notably, the double mutant has a ‘healthy’ phenotype without physiological impairments. Moreover, PaGlo1/PaGlo2_OEx strains are not long-lived on media containing standard glucose concentrations suggesting a tight correlation between the efficiency and capacity to remove MG within the cell, the level of available glucose and lifespan. Overall, our results identify the up-regulation of both components of the glyoxalase system as an effective intervention to increase lifespan in P. anserina. PMID:21212464

  7. Peroxisome dynamics during development of the fungus Podospora anserina.

    PubMed

    Takano-Rojas, Harumi; Zickler, Denise; Peraza-Reyes, Leonardo

    2016-05-01

    Peroxisomes are versatile and dynamic organelles that are required for the development of diverse eukaryotic organisms. We demonstrated previously that in the fungus Podospora anserina different peroxisomal functions are required at distinct stages of sexual development, including the initiation and progression of meiocyte (ascus) development and the differentiation and germination of sexual spores (ascospores). Peroxisome assembly during these processes relies on the differential activity of the protein machinery that drives the import of proteins into the organelle, indicating a complex developmental regulation of peroxisome formation and activity. Here we demonstrate that peroxisome dynamics is also highly regulated during development. We show that peroxisomes in P. anserina are highly dynamic and respond to metabolic and environmental cues by undergoing changes in size, morphology and number. In addition, peroxisomes of vegetative and sexual cell types are structurally different. During sexual development peroxisome number increases at two stages: at early ascus differentiation and during ascospore formation. These processes are accompanied by changes in peroxisome structure and distribution, which include a cell-polarized concentration of peroxisomes at the beginning of ascus development, as well as a morphological transition from predominantly spherical to elongated shapes at the end of the first meiotic division. Further, the mostly tubular peroxisomes present from second meiotic division to early ascospore formation again become rounded during ascospore differentiation. Ultimately the number of peroxisomes dramatically decreases upon ascospore maturation. Our results reveal a precise regulation of peroxisome dynamics during sexual development and suggest that peroxisome constitution and function during development is defined by the coordinated regulation of the proteins that control peroxisome assembly and dynamics. PMID:26908647

  8. Natural Variation of Heterokaryon Incompatibility Gene het-c in Podospora anserina Reveals Diversifying Selection

    PubMed Central

    Bastiaans, Eric; Debets, Alfons J.M.; Aanen, Duur K.; van Diepeningen, Anne D.; Saupe, Sven J.; Paoletti, Mathieu

    2014-01-01

    In filamentous fungi, allorecognition takes the form of heterokaryon incompatibility, a cell death reaction triggered when genetically distinct hyphae fuse. Heterokaryon incompatibility is controlled by specific loci termed het-loci. In this article, we analyzed the natural variation in one such fungal allorecognition determinant, the het-c heterokaryon incompatibility locus of the filamentous ascomycete Podospora anserina. The het-c locus determines an allogenic incompatibility reaction together with two unlinked loci termed het-d and het-e. Each het-c allele is incompatible with a specific subset of the het-d and het-e alleles. We analyzed variability at the het-c locus in a population of 110 individuals, and in additional isolates from various localities. We identified a total of 11 het-c alleles, which define 7 distinct incompatibility specificity classes in combination with the known het-d and het-e alleles. We found that the het-c allorecognition gene of P. anserina is under diversifying selection. We find a highly unequal allele distribution of het-c in the population, which contrasts with the more balanced distribution of functional groups of het-c based on their allorecognition function. One explanation for the observed het-c diversity in the population is its function in allorecognition. However, alleles that are most efficient in allorecognition are rare. An alternative and not exclusive explanation for the observed diversity is that het-c is involved in pathogen recognition. In Arabidopsis thaliana, a homolog of het-c is a pathogen effector target, supporting this hypothesis. We hypothesize that the het-c diversity in P. anserina results from both its functions in pathogen-defense, and allorecognition. PMID:24448643

  9. Natural variation of heterokaryon incompatibility gene het-c in Podospora anserina reveals diversifying selection.

    PubMed

    Bastiaans, Eric; Debets, Alfons J M; Aanen, Duur K; van Diepeningen, Anne D; Saupe, Sven J; Paoletti, Mathieu

    2014-04-01

    In filamentous fungi, allorecognition takes the form of heterokaryon incompatibility, a cell death reaction triggered when genetically distinct hyphae fuse. Heterokaryon incompatibility is controlled by specific loci termed het-loci. In this article, we analyzed the natural variation in one such fungal allorecognition determinant, the het-c heterokaryon incompatibility locus of the filamentous ascomycete Podospora anserina. The het-c locus determines an allogenic incompatibility reaction together with two unlinked loci termed het-d and het-e. Each het-c allele is incompatible with a specific subset of the het-d and het-e alleles. We analyzed variability at the het-c locus in a population of 110 individuals, and in additional isolates from various localities. We identified a total of 11 het-c alleles, which define 7 distinct incompatibility specificity classes in combination with the known het-d and het-e alleles. We found that the het-c allorecognition gene of P. anserina is under diversifying selection. We find a highly unequal allele distribution of het-c in the population, which contrasts with the more balanced distribution of functional groups of het-c based on their allorecognition function. One explanation for the observed het-c diversity in the population is its function in allorecognition. However, alleles that are most efficient in allorecognition are rare. An alternative and not exclusive explanation for the observed diversity is that het-c is involved in pathogen recognition. In Arabidopsis thaliana, a homolog of het-c is a pathogen effector target, supporting this hypothesis. We hypothesize that the het-c diversity in P. anserina results from both its functions in pathogen-defense, and allorecognition. PMID:24448643

  10. Insights into Exo- and Endoglucanase Activities of Family 6 Glycoside Hydrolases from Podospora anserina

    PubMed Central

    Poidevin, Laetitia; Feliu, Julia; Doan, Annick; Berrin, Jean-Guy; Bey, Mathieu; Coutinho, Pedro M.; Henrissat, Bernard; Record, Eric

    2013-01-01

    The ascomycete Podospora anserina is a coprophilous fungus that grows at late stages on droppings of herbivores. Its genome encodes a large diversity of carbohydrate-active enzymes. Among them, four genes encode glycoside hydrolases from family 6 (GH6), the members of which comprise putative endoglucanases and exoglucanases, some of them exerting important functions for biomass degradation in fungi. Therefore, this family was selected for functional analysis. Three of the enzymes, P. anserina Cel6A (PaCel6A), PaCel6B, and PaCel6C, were functionally expressed in the yeast Pichia pastoris. All three GH6 enzymes hydrolyzed crystalline and amorphous cellulose but were inactive on hydroxyethyl cellulose, mannan, galactomannan, xyloglucan, arabinoxylan, arabinan, xylan, and pectin. PaCel6A had a catalytic efficiency on cellotetraose comparable to that of Trichoderma reesei Cel6A (TrCel6A), but PaCel6B and PaCel6C were clearly less efficient. PaCel6A was the enzyme with the highest stability at 45°C, while PaCel6C was the least stable enzyme, losing more than 50% of its activity after incubation at temperatures above 30°C for 24 h. In contrast to TrCel6A, all three studied P. anserina GH6 cellulases were stable over a wide range of pHs and conserved high activity at pH values of up to 9. Each enzyme displayed a distinct substrate and product profile, highlighting different modes of action, with PaCel6A being the enzyme most similar to TrCel6A. PaCel6B was the only enzyme with higher specific activity on carboxymethylcellulose (CMC) than on Avicel and showed lower processivity than the others. Structural modeling predicts an open catalytic cleft, suggesting that PaCel6B is an endoglucanase. PMID:23645193

  11. A Network of HMG-box Transcription Factors Regulates Sexual Cycle in the Fungus Podospora anserina

    PubMed Central

    Ait Benkhali, Jinane; Coppin, Evelyne; Brun, Sylvain; Peraza-Reyes, Leonardo; Martin, Tom; Dixelius, Christina; Lazar, Noureddine; van Tilbeurgh, Herman; Debuchy, Robert

    2013-01-01

    High-mobility group (HMG) B proteins are eukaryotic DNA-binding proteins characterized by the HMG-box functional motif. These transcription factors play a pivotal role in global genomic functions and in the control of genes involved in specific developmental or metabolic pathways. The filamentous ascomycete Podospora anserina contains 12 HMG-box genes. Of these, four have been previously characterized; three are mating-type genes that control fertilization and development of the fruit-body, whereas the last one encodes a factor involved in mitochondrial DNA stability. Systematic deletion analysis of the eight remaining uncharacterized HMG-box genes indicated that none were essential for viability, but that seven were involved in the sexual cycle. Two HMG-box genes display striking features. PaHMG5, an ortholog of SpSte11 from Schizosaccharomyces pombe, is a pivotal activator of mating-type genes in P. anserina, whereas PaHMG9 is a repressor of several phenomena specific to the stationary phase, most notably hyphal anastomoses. Transcriptional analyses of HMG-box genes in HMG-box deletion strains indicated that PaHMG5 is at the hub of a network of several HMG-box factors that regulate mating-type genes and mating-type target genes. Genetic analyses revealed that this network also controls fertility genes that are not regulated by mating-type transcription factors. This study points to the critical role of HMG-box members in sexual reproduction in fungi, as 11 out of 12 members were involved in the sexual cycle in P. anserina. PaHMG5 and SpSte11 are conserved transcriptional regulators of mating-type genes, although P. anserina and S. pombe diverged 550 million years ago. Two HMG-box genes, SOX9 and its upstream regulator SRY, also play an important role in sex determination in mammals. The P. anserina and S. pombe mating-type genes and their upstream regulatory factor form a module of HMG-box genes analogous to the SRY/SOX9 module, revealing a commonality of sex regulation in animals and fungi. PMID:23935511

  12. Age-Related Cellular Copper Dynamics in the Fungal Ageing Model Podospora anserina and in Ageing Human Fibroblasts

    PubMed Central

    Scheckhuber, Christian Q.; Grief, Jürgen; Boilan, Emmanuelle; Luce, Karin; Debacq-Chainiaux, Florence; Rittmeyer, Claudia; Gredilla, Ricardo; Kolbesen, Bernd O.; Toussaint, Olivier; Osiewacz, Heinz D.

    2009-01-01

    In previous investigations an impact of cellular copper homeostasis on ageing of the ascomycete Podospora anserina has been demonstrated. Here we provide new data indicating that mitochondria play a major role in this process. Determination of copper in the cytosolic fraction using total reflection X-ray fluorescence spectroscopy analysis and eGfp reporter gene studies indicate an age-related increase of cytosolic copper levels. We show that components of the mitochondrial matrix (i.e. eGFP targeted to mitochondria) become released from the organelle during ageing. Decreasing the accessibility of mitochondrial copper in P. anserina via targeting a copper metallothionein to the mitochondrial matrix was found to result in a switch from a copper-dependent cytochrome-c oxidase to a copper-independent alternative oxidase type of respiration and results in lifespan extension. In addition, we demonstrate that increased copper concentrations in the culture medium lead to the appearance of senescence biomarkers in human diploid fibroblasts (HDFs). Significantly, expression of copper-regulated genes is induced during in vitro ageing in medium devoid of excess copper suggesting that cytosolic copper levels also increase during senescence of HDFs. These data suggest that the identified molecular pathway of age-dependent copper dynamics may not be restricted to P. anserina but may be conserved from lower eukaryotes to humans. PMID:19305496

  13. Cello-Oligosaccharide Oxidation Reveals Differences between Two Lytic Polysaccharide Monooxygenases (Family GH61) from Podospora anserina

    PubMed Central

    Bey, Mathieu; Zhou, Simeng; Poidevin, Laetitia; Henrissat, Bernard; Coutinho, Pedro M.; Sigoillot, Jean-Claude

    2013-01-01

    The genome of the coprophilic ascomycete Podospora anserina encodes 33 different genes encoding copper-dependent lytic polysaccharide monooxygenases (LPMOs) from glycoside hydrolase family 61 (GH61). In this study, two of these enzymes (P. anserina GH61A [PaGH61A] and PaGH61B), which both harbored a family 1 carbohydrate binding module, were successfully produced in Pichia pastoris. Synergistic cooperation between PaGH61A or PaGH61B with the cellobiose dehydrogenase (CDH) of Pycnoporus cinnabarinus on cellulose resulted in the formation of oxidized and nonoxidized cello-oligosaccharides. A striking difference between PaGH61A and PaGH61B was observed through the identification of the products, among which were doubly and triply oxidized cellodextrins, which were released only by the combination of PaGH61B with CDH. The mass spectrometry fragmentation patterns of these oxidized products could be consistent with oxidation at the C-6 position with a geminal diol group. The different properties of PaGH61A and PaGH61B and their effect on the interaction with CDH are discussed in regard to the proposed in vivo function of the CDH/GH61 enzyme system in oxidative cellulose hydrolysis. PMID:23124232

  14. Altered Mating-Type Identity in the Fungus Podospora Anserina Leads to Selfish Nuclei, Uniparental Progeny, and Haploid Meiosis

    PubMed Central

    Zickler, D.; Arnaise, S.; Coppin, E.; Debuchy, R.; Picard, M.

    1995-01-01

    In wild-type crosses of the filamentous ascomycete Podospora anserina, after fertilization, only nuclei of opposite mating type can form dikaryons that undergo karyogamy and meiosis, producing biparental progeny. To determine the role played by the mating type in these steps, the four mat genes were mutagenized in vitro and introduced into a strain deleted for its mat locus. Genetic and cytological analyses of these mutant strains, crossed to each other and to wild type, showed that mating-type information is required for recognition of nuclear identity during the early steps of sexual reproduction. In crosses with strains carrying a mating-type mutation, two unusual developmental patterns were observed: monokaryotic cells, resulting in haploid meiosis, and uniparental dikaryotic cells providing, after karyogamy and meiosis, a uniparental progeny. Altered mating-type identity leads to selfish behavior of the mutant nucleus: it migrates alone or paired, ignoring its wild-type partner in all mutant X wild-type crosses. This behavior is nucleus-autonomous because, in the same cytoplasm, the wild-type nuclei form only biparental dikaryons. In P. anserina, mat genes are thus required to ensure a biparental dikaryotic state but appear dispensable for later stages, such as meiosis and sporulation. PMID:7498731

  15. Bilirubin oxidase-like proteins from Podospora anserina: promising thermostable enzymes for application in transformation of plant biomass.

    PubMed

    Xie, Ning; Ruprich-Robert, Gwenaël; Silar, Philippe; Chapeland-Leclerc, Florence

    2015-03-01

    Plant biomass degradation by fungi is a critical step for production of biofuels, and laccases are common ligninolytic enzymes envisioned for ligninolysis. Bilirubin oxidases (BODs)-like are related to laccases, but their roles during lignocellulose degradation have not yet been fully investigated. The two BODs of the ascomycete fungus Podospora anserina were characterized by targeted gene deletions. Enzymatic assay revealed that the bod1(Δ) and bod2(Δ) mutants lost partly a thermostable laccase activity. A triple mutant inactivated for bod1, bod2 and mco, a previously investigated multicopper oxidase gene distantly related to laccases, had no thermostable laccase activity. The pattern of fruiting body production in the bod1(Δ) bod2(Δ) double mutant was changed. The bod1(Δ) and bod2(Δ) mutants were reduced in their ability to grow on ligneous and cellulosic materials. Furthermore, bod1(Δ) and bod2(Δ) mutants were defective towards resistance to phenolic substrates and H2 O2 , which may also impact lignocellulose breakdown. Double and triple mutants were more affected than single mutants, evidencing redundancy of function among BODs and mco. Overall, the data show that bod1, bod2 and mco code for non-canonical thermostable laccases that participate in the degradation of lignocellulose. Thanks to their thermal stability, these enzymes may be more promising candidate for biotechnological application than canonical laccases. PMID:24947769

  16. Poly(ADP-Ribose) Polymerase Is a Substrate Recognized by Two Metacaspases of Podospora anserina

    PubMed Central

    Strobel, Ingmar

    2013-01-01

    The two metacaspases MCA1 and MCA2 of the fungal aging model organism Podospora anserina (PaMCA1 and PaMCA2, respectively) have previously been demonstrated to be involved in the control of programmed cell death (PCD) and life span. In order to identify specific pathways and components which are controlled by the activity of these enzymes, we set out to characterize them further. Heterologous overexpression in Escherichia coli of the two metacaspase genes resulted in the production of proteins which aggregate and form inclusion bodies from which the active protein has been recovered via refolding in appropriate buffers. The renaturated proteins are characterized by an arginine-specific activity and are active in caspase-like self-maturation leading to the generation of characteristic small protein fragments. Both activities are dependent on the presence of calcium. Incubation of the two metacaspases with recombinant poly(ADP-ribose) polymerase (PARP), a known substrate of mammalian caspases, led to the identification of PARP as a substrate of the two P. anserina proteases. Using double mutants in which P. anserina Parp (PaParp) is overexpressed and PaMca1 is either overexpressed or deleted, we provide evidence for in vivo degradation of PaPARP by PaMCA1. These results support the idea that the substrate profiles of caspases and metacaspases are at least partially overlapping. Moreover, they link PCD and DNA maintenance in the complex network of molecular pathways involved in aging and life span control. PMID:23584991

  17. Incompatibility in Podospora anserina: Comparative Properties of the Antagonistic Cytoplasmic Factors of a Nonallelic System

    PubMed Central

    Labarère, Jacques; Bégueret, Joël; Bernet, Jean

    1974-01-01

    In the fungus Podospora anserina, the interaction between the nonallelic incompatible R and V genes has two consequences: a lytic reaction due to the synthesis of specific proteolytic enzymes, and a quenching in protein and ribonucleic acid synthesis. The incompatibility reaction when vegetative or sexual R and V cells fuse is asymmetric: it is induced only in the R protoplasm. The cessation in ribonucleic acid and protein synthesis was investigated in heterokaryotic strains carrying the antagonistic R and V genes and their “neutral” r and v alleles. Asymmetry between R and V genes lies in the fact that the strains homozygous for the R genes are the only strains that cannot grow. From these results it is postulated that the V-gene product is a diffusible cytoplasmic factor and that the R-gene product, which is nonautonomous, is a ribosomal component. PMID:4142030

  18. A unique CE16 acetyl esterase from Podospora anserina active on polymeric xylan.

    PubMed

    Puchart, Vladimír; Berrin, Jean-Guy; Haon, Mireille; Biely, Peter

    2015-12-01

    The genome of the coprophilous fungus Podospora anserina displays an impressive array of genes encoding hemicellulolytic enzymes. In this study, we focused on a putative carbohydrate esterase (CE) from family 16 (CE16) that bears a carbohydrate-binding module from family CBM1. The protein was heterologously expressed in Pichia pastoris and purified to electrophoretic homogeneity. The P. anserina CE16 enzyme (PaCE16A) exhibited different catalytic properties than so far known CE16 esterases represented by the Trichoderma reesei CE16 acetyl esterase (TrCE16). A common property of both CE16 esterases is their exodeacetylase activity, i.e., deesterification at positions 3 and 4 of monomeric xylosides and the nonreducing end xylopyranosyl (Xylp) residue of oligomeric homologues. However, the PaCE16A showed lower positional specificity than TrCE16 and efficiently deacetylated also position 2. The major difference observed between PaCE16A and TrCE16 was found on polymeric substrate, acetylglucuronoxylan. While TrCE16 does not attack internal acetyl groups, PaCE16A deacetylated singly and doubly acetylated Xylp residues in the polymer to such an extent that it resulted in the polymer precipitation. Similarly as typical acetylxylan esterases belonging to CE1, CE4, CE5, and CE6 families, PaCE16A did not attack 3-O-acetyl group of xylopyranosyl residues carrying 4-O-methyl-D-glucuronic acid at position 2. PaCE16A thus represents a CE16 member displaying unique catalytic properties, which are intermediate between the TrCE16 exodeacetylase and acetylxylan esterases designed to deacetylate polymeric substrate. The catalytic versatility of PaCE16A makes the enzyme an important candidate for biotechnological applications. PMID:26329850

  19. Enzymatic synthesis of model substrates recognized by glucuronoyl esterases from Podospora anserina and Myceliophthora thermophila.

    PubMed

    Katsimpouras, Constantinos; Bénarouche, Anaïs; Navarro, David; Karpusas, Michael; Dimarogona, Maria; Berrin, Jean-Guy; Christakopoulos, Paul; Topakas, Evangelos

    2014-06-01

    Glucuronoyl esterases (GEs) are recently discovered enzymes that are suggested to cleave the ester bond between lignin alcohols and xylan-bound 4-O-methyl-D-glucuronic acid. Although their potential use for enhanced enzymatic biomass degradation and synthesis of valuable chemicals renders them attractive research targets for biotechnological applications, the difficulty to purify natural fractions of lignin-carbohydrate complexes hampers the characterization of fungal GEs. In this work, we report the synthesis of three aryl alkyl or alkenyl D-glucuronate esters using lipase B from Candida antarctica (CALB) and their use to determine the kinetic parameters of two GEs, StGE2 from the thermophilic fungus Myceliophthora thermophila (syn. Sporotrichum thermophile) and PaGE1 from the coprophilous fungus Podospora anserina. PaGE1 was functionally expressed in the methylotrophic yeast Pichia pastoris under the transcriptional control of the alcohol oxidase (AOX1) promoter and purified to its homogeneity (63 kDa). The three D-glucuronate esters contain an aromatic UV-absorbing phenol group that facilitates the quantification of their enzymatic hydrolysis by HPLC. Both enzymes were able to hydrolyze the synthetic esters with a pronounced preference towards the cinnamyl-D-glucuronate ester. The experimental results were corroborated by computational docking of the synthesized substrate analogues. We show that the nature of the alcohol portion of the hydrolyzed ester influences the catalytic efficiency of the two GEs. PMID:24531271

  20. Identification of a Hypothetical Protein from Podospora anserina as a Nitroalkane Oxidase

    SciTech Connect

    Tormos, Jose R.; Taylor, Alexander B.; Daubner, S. Colette; Hart, P. John; Fitzpatrick, Paul F.

    2010-08-23

    The flavoprotein nitroalkane oxidase (NAO) from Fusarium oxysporum catalyzes the oxidation of primary and secondary nitroalkanes to their respective aldehydes and ketones. Structurally, the enzyme is a member of the acyl-CoA dehydrogenase superfamily. To date no enzymes other than that from F. oxysporum have been annotated as NAOs. To identify additional potential NAOs, the available database was searched for enzymes in which the active site residues Asp402, Arg409, and Ser276 were conserved. Of the several fungal enzymes identified in this fashion, PODANSg2158 from Podospora anserina was selected for expression and characterization. The recombinant enzyme is a flavoprotein with activity on nitroalkanes comparable to the F. oxysporum NAO, although the substrate specificity is somewhat different. Asp399, Arg406, and Ser273 in PODANSg2158 correspond to the active site triad in F. oxysporum NAO. The k{sub cat}/K{sub M}-pH profile with nitroethane shows a pK{sub a} of 5.9 that is assigned to Asp399 as the active site base. Mutation of Asp399 to asparagine decreases the k{sub cat}/K{sub M} value for nitroethane over 2 orders of magnitude. The R406K and S373A mutations decrease this kinetic parameter by 64- and 3-fold, respectively. The structure of PODANSg2158 has been determined at a resolution of 2.0 {angstrom}, confirming its identification as an NAO.

  1. Overlapping Podospora anserina Transcriptional Responses to Bacterial and Fungal Non Self Indicate a Multilayered Innate Immune Response

    PubMed Central

    Lamacchia, Marina; Dyrka, Witold; Breton, Annick; Saupe, Sven J.; Paoletti, Mathieu

    2016-01-01

    Recognition and response to non self is essential to development and survival of all organisms. It can occur between individuals of the same species or between different organisms. Fungi are established models for conspecific non self recognition in the form of vegetative incompatibility (VI), a genetically controlled process initiating a programmed cell death (PCD) leading to the rejection of a fusion cell between genetically different isolates of the same species. In Podospora anserina VI is controlled by members of the hnwd gene family encoding for proteins analogous to NOD Like Receptors (NLR) immune receptors in eukaryotes. It was hypothesized that the hnwd controlled VI reaction was derived from the fungal innate immune response. Here we analyze the P. anserina transcriptional responses to two bacterial species, Serratia fonticola to which P. anserina survives and S. marcescens to which P. anserina succumbs, and compare these to the transcriptional response induced under VI conditions. Transcriptional responses to both bacteria largely overlap, however the number of genes regulated and magnitude of regulation is more important when P. anserina survives. Transcriptional responses to bacteria also overlap with the VI reaction for both up or down regulated gene sets. Genes up regulated tend to be clustered in the genome, and display limited phylogenetic distribution. In all three responses we observed genes related to autophagy to be up-regulated. Autophagy contributes to the fungal survival in all three conditions. Genes encoding for secondary metabolites and histidine kinase signaling are also up regulated in all three conditions. Transcriptional responses also display differences. Genes involved in response to oxidative stress, or encoding small secreted proteins are essentially expressed in response to bacteria, while genes encoding NLR proteins are expressed during VI. Most functions encoded in response to bacteria favor survival of the fungus while most functions up regulated during VI would lead to cell death. These differences are discussed in the frame of a multilayered response to non self in fungi. PMID:27148175

  2. Incomplete penetrance and variable expressivity of a growth defect as a consequence of knocking out two K(+) transporters in the euascomycete fungus Podospora anserina.

    PubMed Central

    Lalucque, Herv; Silar, Philippe

    2004-01-01

    We describe an example of incomplete penetrance and variable expressivity in the filamentous fungus Podospora anserina, two genetic properties classically associated with mutations in more complex organisms, such as green plants and animals. We show that the knockouts of two TRK-related K(+) transporters of this ascomycete present variability in their phenotype that cannot be attributed to fluctuations of the genetic background or the environment. Thalli of the knockout strains derived from independent monokaryotic ascospores or from a single monokaryotic ascospore and cultivated under standard growth conditions may or may not present impaired growth. When impaired, thalli exhibit a range of phenotypes. Environmental conditions control expressivity to a large extent and penetrance to a low extent. Restoration of functional potassium transport by heterologous expression of K(+) transporters from Neurospora crassa abolishes or strongly diminishes the growth impairment. These data show that incomplete penetrance and variable expressivity can be an intrinsic property of a single Mendelian loss-of-function mutation. They also show that such variability in the expression of a mutant phenotype can be promoted by a phenomenon not obviously related to the well-known chromatin structure modifications, i.e., potassium transport. They provide a framework to understand human channelopathies with similar properties. PMID:15020412

  3. Altering a gene involved in nuclear distribution increases the repeat-induced point mutation process in the fungus Podospora anserina.

    PubMed Central

    Bouhouche, Khaled; Zickler, Denise; Debuchy, Robert; Arnaise, Sylvie

    2004-01-01

    Repeat-induced point mutation (RIP) is a homology-dependent gene-silencing mechanism that introduces C:G-to-T:A transitions in duplicated DNA segments. Cis-duplicated sequences can also be affected by another mechanism called premeiotic recombination (PR). Both are active over the sexual cycle of some filamentous fungi, e.g., Neurospora crassa and Podospora anserina. During the sexual cycle, several developmental steps require precise nuclear movement and positioning, but connections between RIP, PR, and nuclear distributions have not yet been established. Previous work has led to the isolation of ami1, the P. anserina ortholog of the Aspergillus nidulans apsA gene, which is required for nuclear positioning. We show here that ami1 is involved in nuclear distribution during the sexual cycle and that alteration of ami1 delays the fruiting-body development. We also demonstrate that ami1 alteration affects loss of transgene functions during the sexual cycle. Genetically linked multiple copies of transgenes are affected by RIP and PR much more frequently in an ami1 mutant cross than in a wild-type cross. Our results suggest that the developmental slowdown of the ami1 mutant during the period of RIP and PR increases time exposure to the duplication detection system and thus increases the frequency of RIP and PR. PMID:15166143

  4. The HET-s prion protein of the filamentous fungus Podospora anserina aggregates in vitro into amyloid-like fibrils.

    PubMed

    Dos Reis, Suzana; Coulary-Salin, Bénédicte; Forge, Vincent; Lascu, Ioan; Bégueret, Joël; Saupe, Sven J

    2002-02-22

    The HET-s protein of Podospora anserina is a fungal prion. This protein behaves as an infectious cytoplasmic element that is transmitted horizontally from one strain to another. Under the prion form, the HET-s protein forms aggregates in vivo. The specificity of this prion model compared with the yeast prions resides in the fact that under the prion form HET-s causes a growth inhibition and cell death reaction when co-expressed with the HET-S protein from which it differs by 13 residues. Herein we describe the purification and initial characterization of recombinant HET-s protein expressed in Escherichia coli. The HET-s protein self-associates over time into high molecular weight aggregates. These aggregates greatly accelerate precipitation of the soluble form. HET-s aggregates appear as amyloid-like fibrils using electron microscopy. They bind Congo Red and show birefringence under polarized light. In the aggregated form, a HET-s fragment of approximately 7 kDa is resistant to proteinase K digestion. CD and FTIR analyses indicate that upon transition to the aggregated state, the HET-s protein undergoes a structural rearrangement characterized by an increase in antiparallel beta-sheet structure content. These results suggest that the [Het-s] prion element propagates in vivo as an infectious amyloid. PMID:11733532

  5. Co-expression of the mating-type genes involved in internuclear recognition is lethal in Podospora anserina.

    PubMed Central

    Coppin, E; Debuchy, R

    2000-01-01

    In the heterothallic filamentous fungus Podospora anserina, four mating-type genes encoding transcriptional factors have been characterized: FPR1 in the mat+ sequence and FMR1, SMR1, and SMR2 in the alternative mat- sequence. Fertilization is controlled by FPR1 and FMR1. After fertilization, male and female nuclei, which have divided in the same cell, form mat+/mat- pairs during migration into the ascogenous hyphae. Previous data indicate that the formation of mat+/mat- pairs is controlled by FPR1, FMR1, and SMR2. SMR1 was postulated to be necessary for initial development of ascogenous hyphae. In this study, we investigated the transcriptional control of the mat genes by seeking mat transcripts during the vegetative and sexual phase and fusing their promoter to a reporter gene. The data indicate that FMR1 and FPR1 are expressed in both mycelia and perithecia, whereas SMR1 and SMR2 are transcribed in perithecia. Increased or induced vegetative expression of the four mat genes has no effect when the recombined gene is solely in the wild-type strain. However, the combination of resident FPR1 with deregulated SMR2 and overexpressed FMR1 in the same nucleus is lethal. This lethality is suppressed by the expression of SMR1, confirming that SMR1 operates downstream of the other mat genes. PMID:10835389

  6. Genetic and functional investigation of Zn(2)Cys(6) transcription factors RSE2 and RSE3 in Podospora anserina.

    PubMed

    Bovier, Elodie; Sellem, Carole H; Humbert, Adeline; Sainsard-Chanet, Annie

    2014-01-01

    In Podospora anserina, the two zinc cluster proteins RSE2 and RSE3 are essential for the expression of the gene encoding the alternative oxidase (aox) when the mitochondrial electron transport chain is impaired. In parallel, they activated the expression of gluconeogenic genes encoding phosphoenolpyruvate carboxykinase (pck) and fructose-1,6-biphosphatase (fbp). Orthologues of these transcription factors are present in a wide range of filamentous fungi, and no other role than the regulation of these three genes has been evidenced so far. In order to better understand the function and the organization of RSE2 and RSE3, we conducted a saturated genetic screen based on the constitutive expression of the aox gene. We identified 10 independent mutations in 9 positions in rse2 and 11 mutations in 5 positions in rse3. Deletions were generated at some of these positions and the effects analyzed. This analysis suggests the presence of central regulatory domains and a C-terminal activation domain in both proteins. Microarray analysis revealed 598 genes that were differentially expressed in the strains containing gain- or loss-of-function mutations in rse2 or rse3. It showed that in addition to aox, fbp, and pck, RSE2 and RSE3 regulate the expression of genes encoding the alternative NADH dehydrogenase, a Zn2Cys6 transcription factor, a flavohemoglobin, and various hydrolases. As a complement to expression data, a metabolome profiling approach revealed that both an rse2 gain-of-function mutation and growth on antimycin result in similar metabolic alterations in amino acids, fatty acids, and α-ketoglutarate pools. PMID:24186951

  7. Genome-Wide Gene Expression Profiling of Fertilization Competent Mycelium in Opposite Mating Types in the Heterothallic Fungus Podospora anserina

    PubMed Central

    Coppin, Evelyne; Imbeaud, Sandrine; Grognet, Pierre; Delacroix, Hervé; Debuchy, Robert

    2011-01-01

    Background Mating-type loci in yeasts and ascomycotan filamentous fungi (Pezizomycotina) encode master transcriptional factors that play a critical role in sexual development. Genome-wide analyses of mating-type-specification circuits and mating-type target genes are available in Saccharomyces cerevisiae and Schizosaccharomyces pombe; however, no such analyses have been performed in heterothallic (self-incompatible) Pezizomycotina. The heterothallic fungus Podospora anserina serves as a model for understanding the basic features of mating-type control. Its mat+ and mat− mating types are determined by dissimilar allelic sequences. The mat− sequence contains three genes, designated FMR1, SMR1 and SMR2, while the mat+ sequence contains one gene, FPR1. FMR1 and FPR1 are the major regulators of fertilization, and this study presents a genome-wide view of their target genes and analyzes their target gene regulation. Methodology/Principal Findings The transcriptomic profiles of the mat+ and mat− strains revealed 157 differentially transcribed genes, and transcriptomic analysis of fmr1− and fpr1− mutant strains was used to determine the regulatory actions exerted by FMR1 and FPR1 on these differentially transcribed genes. All possible combinations of transcription repression and/or activation by FMR1 and/or FPR1 were observed. Furthermore, 10 additional mating-type target genes were identified that were up- or down-regulated to the same level in mat+ and mat− strains. Of the 167 genes identified, 32 genes were selected for deletion, which resulted in the identification of two genes essential for the sexual cycle. Interspecies comparisons of mating-type target genes revealed significant numbers of orthologous pairs, although transcriptional profiles were not conserved between species. Conclusions/Significance This study represents the first comprehensive genome-wide analysis of mating-type direct and indirect target genes in a heterothallic filamentous fungus. Mating-type transcription factors have many more target genes than are found in yeasts and exert a much greater diversity of regulatory actions on target genes, most of which are not directly related to mating. PMID:21738678

  8. Deletion of the MED13 and CDK8 subunits of the Mediator improves the phenotype of a long-lived respiratory deficient mutant of Podospora anserina.

    PubMed

    Humbert, Adeline; Bovier, Elodie; Sellem, Carole H; Sainsard-Chanet, Annie

    2015-09-01

    In Podospora anserina, the loss of function of the cytochrome segment of the mitochondrial respiratory chain is viable. This is due to the presence in this organism, as in most filamentous fungi, of an alternative respiratory oxidase (AOX) that provides a bypass to the cytochrome pathway. However mutants lacking a functional cytochrome pathway present multiple phenotypes including poorly colored thin mycelium and slow growth. In a large genetic screen based on the improvement of these phenotypes, we isolated a large number of independent suppressor mutations. Most of them led to the constitutive overexpression of the aox gene. In this study, we characterize a new suppressor mutation that does not affect the production of AOX. It is a loss-of-function mutation in the gene encoding the MED13 subunit of the kinase module of the Mediator complex. Inactivation of the cdk8 gene encoding another subunit of the same module also results in partial suppression of a cytochrome-deficient mutant. Analysis of strains lacking the MED13 or CDK8 subunits points to the importance of these subunits as regulators involved in diverse physiological processes such as growth, longevity and sexual development. Interestingly, transcriptional analyses indicate that in P. anserina, loss of the respiratory cytochrome pathway results in the up-regulation of glycolysis-related genes revealing a new type of retrograde regulation. The loss of MED13 augments the up-regulation of some of these genes. PMID:26231682

  9. The mod-A suppressor of nonallelic heterokaryon incompatibility in Podospora anserina encodes a proline-rich polypeptide involved in female organ formation.

    PubMed Central

    Barreau, C; Iskandar, M; Loubradou, G; Levallois, V; Bégueret, J

    1998-01-01

    Vegetative incompatibility in fungi results from the control of heterokaryon formation by the genes present at het loci. Coexpression of antagonistic het genes in the same hyphae leads to a lethal process. In Podospora anserina, self-incompatible strains containing nonallelic incompatible genes in the same nucleus are inviable as the result of a growth arrest and a lytic process. Mutations in suppressor genes (mod genes) can restore the viability. These mod mutations also interfere with developmental processes, which suggests common steps between the incompatibility reaction and cellular differentiation. The mod-A locus, responsible for growth arrest in the self-incompatible strains, is also involved in the control of the development of female organs. The mod-A gene was isolated. An open reading frame 687 amino acids long was identified. The MOD-A-encoded polypeptide is rich in proline residues, which are clustered in a domain containing a motif that displays similarity to SH3-binding motifs, which are known to be involved in protein-protein interactions. Construction of a strain deleted for mod-A confirmed that the product of this gene involved in differentiation is a key regulator of growth arrest associated with vegetative incompatibility. PMID:9611202

  10. ami1, an orthologue of the Aspergillus nidulans apsA gene, is involved in nuclear migration events throughout the life cycle of Podospora anserina.

    PubMed Central

    Graïa, F; Berteaux-Lecellier, V; Zickler, D; Picard, M

    2000-01-01

    The Podospora anserina ami1-1 mutant was identified as a male-sterile strain. Microconidia (which act as male gametes) form, but are anucleate. Paraphysae from the perithecium beaks are also anucleate when ami1-1 is used as the female partner in a cross. Furthermore, in crosses heterozygous for ami1-1, some crozier cells are uninucleate rather than binucleate. In addition to these nuclear migration defects, which occur at the transition between syncytial and cellular states, ami1-1 causes abnormal distribution of the nuclei in both mycelial filaments and asci. Finally, an ami1-1 strain bearing information for both mating types is unable to self-fertilize. The ami1 gene is an orthologue of the Aspergillus nidulans apsA gene, which controls nuclear positioning in filaments and during conidiogenesis (at the syncytial/cellular transition). The ApsA and AMI1 proteins display 42% identity and share structural features. The apsA gene complements some ami1-1 defects: it increases the percentage of nucleate microconidia and restores self-fertility in an ami1-1 mat+ (mat-) strain. The latter effect is puzzling, since in apsA null mutants sexual reproduction is quite normal. The functional differences between the two genes are discussed with respect to their possible history in these two fungi, which are very distant in terms of evolution. PMID:10835387

  11. A New Method for the Screening of Mutations Related to Protoplasmic Incompatibility, Differentiation and Plasma Membrane in the Fungus PODOSPORA ANSERINA

    PubMed Central

    Labarère, Jacques; Bonneu, Marc

    1985-01-01

    In Podospora anserina previous investigations showed that mutations in genes involved in the control of protoplasmic incompatibility cause defects at various stages of differentiation during the life cycle and also modify properties of the plasma membrane. To establish these relationships in another way, a new method for screening mutations has been developed as a first step. Eighty-five new mutants were selected for resistance to toxic products (sorbose or thiourea); in a second step these mutants were tested for modifications of protoplasmic incompatibility and cellular differentiation. Seven of the sorbose or thiourea resistant-mutants (i.e., 8%) exhibited new patterns of protoplasmic incompatibility. Genetic analyses were carried out with three mutants. Mutation X25 suppresses protoplasmic incompatibility resulting from all allelic interactions and restores the fertility of the crosses ♀ V x ♂ V1 and ♀ Z1 x ♂ Z2. Mutation V41 is an allele of the v locus with new properties. Mutation X61 totally suppresses the V/V'1 interaction and weakens all of the other allelic incompatibility systems; X61 strains are defective in protoperithecia differentiation. Electrophoresis of plasma membrane proteins showed that X61 strains lack two polypeptides whose apparent molecular weights are 41,000 and 44,000. This new screening method is especially efficient for obtaining new mutants and identifying additional genes involved in incompatibility. These results provide further support demonstrating the relationships between protoplasmic incompatibility, cellular differentiation and plasma membrane. PMID:17246298

  12. eEF1A Controls ascospore differentiation through elevated accuracy, but controls longevity and fruiting body formation through another mechanism in Podospora anserina.

    PubMed Central

    Silar, P; Lalucque, H; Haedens, V; Zickler, D; Picard, M

    2001-01-01

    Antisuppressor mutations in the eEF1A gene of Podospora anserina were previously shown to impair ascospore formation, to drastically increase life span, and to permit the development of the Crippled Growth degenerative process. Here, we show that eEF1A controls ascospore formation through accuracy level maintenance. Examination of antisuppressor mutant perithecia reveals two main cytological defects, mislocalization of spindle and nuclei and nuclear death. Antisuppression levels are shown to be highly dependent upon both the mutation site and the suppressor used, precluding any correlation between antisuppression efficiency and severity of the sporulation impairment. Nevertheless, severity of ascospore differentiation defect is correlated with resistance to paromomycin. We also show that eEF1A controls fruiting body formation and longevity through a mechanism(s) different from accuracy control. In vivo, GFP tagging of the protein in a way that partly retains its function confirmed earlier cytological observation; i.e., this factor is mainly diffuse within the cytosol, but may transiently accumulate within nuclei or in defined regions of the cytoplasm. These data emphasize the fact that the translation apparatus exerts a global regulatory control over cell physiology and that eEF1A is one of the key factors involved in this monitoring. PMID:11514440

  13. Identification of the het-r vegetative incompatibility gene of Podospora anserina as a member of the fast evolving HNWD gene family.

    PubMed

    Chevanne, Damien; Bastiaans, Eric; Debets, Alfons; Saupe, Sven J; Clavé, Corinne; Paoletti, Mathieu

    2009-02-01

    In fungi, vegetative incompatibility is a conspecific non-self recognition mechanism that restricts formation of viable heterokaryons when incompatible alleles of specific het loci interact. In Podospora anserina, three non-allelic incompatibility systems have been genetically defined involving interactions between het-c and het-d, het-c and het-e, het-r and het-v. het-d and het-e are paralogues belonging to the HNWD gene family that encode proteins of the STAND class. HET-D and HET-E proteins comprise an N-terminal HET effector domain, a central GTP binding site and a C-terminal WD repeat domain constituted of tandem repeats of highly conserved WD40 repeat units that define the specificity of alleles during incompatibility. The WD40 repeat units of the members of this HNWD family are undergoing concerted evolution. By combining genetic analysis and gain of function experiments, we demonstrate that an additional member of this family, HNWD2, corresponds to the het-r non-allelic incompatibility gene. As for het-d and het-e, allele specificity at the het-r locus is determined by the WD repeat domain. Natural isolates show allelic variation for het-r. PMID:19137300

  14. A homologue of the yeast SHE4 gene is essential for the transition between the syncytial and cellular stages during sexual reproduction of the fungus Podospora anserina.

    PubMed Central

    Berteaux-Lecellier, V; Zickler, D; Debuchy, R; Panvier-Adoutte, A; Thompson-Coffe, C; Picard, M

    1998-01-01

    The Podospora anserina cro1 gene was identified as a gene required for sexual sporulation. Crosses homozygous for the cro1-1 mutation yield fruiting bodies which produce few asci due to the formation of giant plurinucleate cells instead of dikaryotic cells after fertilization. This defect does not impair karyogamy, but meioses of the resultant polyploid nuclei are most often abortive. Cytological studies suggest that the primary defect of the mutant is its inability to form septa between the daughter nuclei after each mitosis, a step specific for normal dikaryotic cell divisions. The cro1-1 mutant would thus be unable to leave the syncytial vegetative state while abiding by the meiotic programme. cro1-1 also shows defects in ascospore germination and growth rate. GFP-tagging of the CRO1 protein reveals that it is a cytosolic protein mainly expressed at the beginning of the dikaryotic stage and at the time of ascospore maturation. The CRO1 protein exhibits significant similarity to the SHE4 protein, which is required for asymmetric mating-type switching in budding yeast cells. Thus, a gene involved in asymmetric cell divisions in a unicellular organism plays a key role at the transition between the syncytial (vegetative) state and the cellular (sexual) state in a filamentous fungus. PMID:9482722

  15. The protein product of the het-s heterokaryon incompatibility gene of the fungus Podospora anserina behaves as a prion analog

    PubMed Central

    Coustou, Virginie; Deleu, Carol; Saupe, Sven; Begueret, Joel

    1997-01-01

    The het-s locus of Podospora anserina is a heterokaryon incompatibility locus. The coexpression of the antagonistic het-s and het-S alleles triggers a lethal reaction that prevents the formation of viable heterokaryons. Strains that contain the het-s allele can display two different phenotypes, [Het-s] or [Het-s*], according to their reactivity in incompatibility. The detection in these phenotypically distinct strains of a protein expressed from the het-s gene indicates that the difference in reactivity depends on a posttranslational difference between two forms of the polypeptide encoded by the het-s gene. This posttranslational modification does not affect the electrophoretic mobility of the protein in SDS/PAGE. Several results suggest a similarity of behavior between the protein encoded by the het-s gene and prions. The [Het-s] character can propagate in [Het-s*] strains as an infectious agent, producing a [Het-s*] → [Het-s] transition, independently of protein synthesis. Expression of the [Het-s] character requires a functional het-s gene. The protein present in [Het-s] strains is more resistant to proteinase K than that present in [Het-s*] mycelium. Furthermore, overexpression of the het-s gene increases the frequency of the transition from [Het-s*] to [Het-s]. We propose that this transition is the consequence of a self-propagating conformational modification of the protein mediated by the formation of complexes between the two different forms of the polypeptide. PMID:9275200

  16. Molecular Genetics of Heterokaryon Incompatibility in Filamentous Ascomycetes

    PubMed Central

    Saupe, Sven J.

    2000-01-01

    Filamentous fungi spontaneously undergo vegetative cell fusion events within but also between individuals. These cell fusions (anastomoses) lead to cytoplasmic mixing and to the formation of vegetative heterokaryons (i.e., cells containing different nuclear types). The viability of these heterokaryons is genetically controlled by specific loci termed het loci (for heterokaryon incompatibility). Heterokaryotic cells formed between individuals of unlike het genotypes undergo a characteristic cell death reaction or else are severely inhibited in their growth. The biological significance of this phenomenon remains a puzzle. Heterokaryon incompatibility genes have been proposed to represent a vegetative self/nonself recognition system preventing heterokaryon formation between unlike individuals to limit horizontal transfer of cytoplasmic infectious elements. Molecular characterization of het genes and of genes participating in the incompatibility reaction has been achieved for two ascomycetes, Neurospora crassa and Podospora anserina. These analyses have shown that het genes are diverse in sequence and do not belong to a gene family and that at least some of them perform cellular functions in addition to their role in incompatibility. Divergence between the different allelic forms of a het gene is generally extensive, but single-amino-acid differences can be sufficient to trigger incompatibility. In some instances het gene evolution appears to be driven by positive selection, which suggests that the het genes indeed represent recognition systems. However, work on nonallelic incompatibility systems in P. anserina suggests that incompatibility might represent an accidental activation of a cellular system controlling adaptation to starvation. PMID:10974123

  17. Mating types and sexual development in filamentous ascomycetes.

    PubMed Central

    Coppin, E; Debuchy, R; Arnaise, S; Picard, M

    1997-01-01

    The progress made in the molecular characterization of the mating types in several filamentous ascomycetes has allowed us to better understand their role in sexual development and has brought to light interesting biological problems. The mating types of Neurospora crassa, Podospora anserina, and Cochliobolus heterostrophus consist of unrelated and unique sequences containing one or several genes with multiple functions, related to sexuality or not, such as vegetative incompatibility in N. crassa. The presence of putative DNA binding domains in the proteins encoded by the mating-type (mat) genes suggests that they may be transcriptional factors. The mat genes play a role in cell-cell recognition at fertilization, probably by activating the genes responsible for the hormonal signal whose occurrence was previously demonstrated by physiological experiments. They also control recognition between nuclei at a later stage, when reproductive nuclei of each mating type which have divided in the common cytoplasm pair within the ascogenous hyphae. How self is distinguished from nonself at the nuclear level is not known. The finding that homothallic species, able to mate in the absence of a partner, contain both mating types in the same haploid genome has raised more issues than it has resolved. The instability of the mating type, in particular in Sclerotinia trifolorium and Botrytinia fuckeliana, is also unexplained. This diversity of mating systems, still more apparent if the yeasts and the basidiomycetes are taken into account, clearly shows that no single species can serve as a universal mating-type model. PMID:9409146

  18. Podospora anserina mutant defective in protoperithecium formation, ascospore germination, and cell regeneration.

    PubMed Central

    Durrens, P; Laigret, F; Labarère, J; Bernet, J

    1979-01-01

    A mutant (modx) was selected on the basis of the suppression of self-lysis due to a recessive mutation (modB). modx, a dominant mutation, reduced hyphal branching from nonapical cells, abolished protoperithecium formation, and induced the death of stationary cells only when these were isolated to obtain further development. Mutant ascospores, formed in the fruiting bodies which occasionally occur under specific conditions (32 degrees C on starved medium), showed a delay in the germination process (up to 3 months instead of about 5 h for wild-type ascospores) when submitted to incubation under standard conditions (26 degrees C on germination medium) and failed to germinate at 18 degrees C. Revertants from modx strains, selected on the basis of the suppression of the nonrenewal of growth from stationary cells, were wild type for all the other three defects. Indirect arguments suggested that the modx mutant strain might be defective in the control of a specific class of stable messenger ribonucleic acids which would be essential for the physiology of ascospores and stationary cells. Images PMID:118158

  19. Protoplasmic Incompatibility in PODOSPORA ANSERINA: a Possible Function for Incompatibility Genes

    PubMed Central

    Boucherie, Hélian; Bernet, Jean

    1980-01-01

    The suppression of protoplasmic incompatibility resulting from nonallelic gene interactions has been obtained by the coupled effect of mutations in the modA and modB genes (Bernet 1971). Due to their female sterility, modA modB strains provide an experimental tool to determine whether or not the mod and incompatibility loci are involved in a function other than protoplasmic incompatibility. Present results show that modA modB female sterility is a nonautonomous trait since heterokaryotic mycelia that include a modA modB nucleus and a female fertile nucleus (wild-type, modA or modB) produce modA modB protoperithecia, which are also formed by culture on medium supplemented with specific amino acids. Using modA modB strains, which are sterile at 32° and fertile at 26°, we have shown that the mod genes have no specific sequential timing. Indeed, the mod mutations may prevent the achievement of the female sexual cycle at any developmental stage from before early differentiation of protoperithecia until ascospore maturation. Employing different modA and modB mutations, we have shown that protoperithecia in modA modB cultures are generally distributed in female fertile rings; this result indicates that protoperithecia occur only in mycelial areas that have a restricted range of age at the time that modA modB thalli complete growth. Furthermore, nonsense mutations of incompatibility genes suppress the modA modB female fertile rings or restrict their width, suggesting that incompatibility loci, like the mod loci, are involved in protoperithecium formation. Taken together, these results lead to the postulate that mod and incompatibility genes do not determine, sensu stricto, protoperithecial function, as previously supposed (Boucherie and Bernet 1974), but may be involved in the homeostatic control of stationary cell functions essential for the complete development of the female sexual cycle. PMID:17249066

  20. The Podospora rmp1 gene implicated in nucleus-mitochondria cross-talk encodes an essential protein whose subcellular location is developmentally regulated.

    PubMed Central

    Contamine, Véronique; Zickler, Denise; Picard, Marguerite

    2004-01-01

    It has been previously reported that, at the time of death, the Podospora anserina AS1-4 mutant strains accumulate specific deleted forms of the mitochondrial genome and that their life spans depend on two natural alleles (variants) of the rmp1 gene: AS1-4 rmp1-2 strains exhibit life spans strikingly longer than those of AS1-4 rmp1-1. Here, we show that rmp1 is an essential gene. In silico analyses of eight rmp1 natural alleles present in Podospora isolates and of the putative homologs of this orphan gene in other filamentous fungi suggest that rmp1 evolves rapidly. The RMP1 protein is localized in the mitochondrial and/or the cytosolic compartment, depending on cell type and developmental stage. Strains producing RMP1 without its mitochondrial targeting peptide are viable but exhibit vegetative and sexual defects. PMID:15020413

  1. A site-specific deletion in mitochondrial DNA of Podospora is under the control of nuclear genes.

    PubMed

    Belcour, L; Begel, O; Picard, M

    1991-05-01

    In the filamentous fungus Podospora anserina, the association of two nuclear genes inevitably leads to a "premature death" phenotype consisting of an early end of vegetative growth a few days after ascospore germination. Mycelia showing this phenotype contain a mitochondrial chromosome that always bears the same deletion. One of the break points is exactly at the 5' splice site of a particular mitochondrial intron, suggesting that the deletion event could result from molecular mechanisms also involved in intron mobility. One of the nuclear genes involved in triggering this site-specific event belongs to the mating-type minus haplotype; the other is a mutant allele of a gene encoding a cytosolic ribosomal protein. PMID:2023905

  2. Rab-GDI Complex Dissociation Factor Expressed through Translational Frameshifting in Filamentous Ascomycetes

    PubMed Central

    Prigent, Magali; Rousset, Jean-Pierre; Namy, Olivier; Silar, Philippe

    2013-01-01

    In the model fungus Podospora anserina, the PaYIP3 gene encoding the orthologue of the Saccharomyces cerevisiae YIP3 Rab-GDI complex dissociation factor expresses two polypeptides, one of which, the long form, is produced through a programmed translation frameshift. Inactivation of PaYIP3 results in slightly delayed growth associated with modification in repartition of fruiting body on the thallus, along with reduced ascospore production on wood. Long and short forms of PaYIP3 are expressed in the mycelium, while only the short form appears expressed in the maturing fruiting body (perithecium). The frameshift has been conserved over the evolution of the Pezizomycotina, lasting for over 400 million years, suggesting that it has an important role in the wild. PMID:24069231

  3. Evolutionary history of Ascomyceteous Yeasts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yeasts are important for many industrial and biotechnological processes and show remarkable diversity despite morphological similarities. We have sequenced the genomes of 20 ascomyceteous yeasts of taxonomic and industrial importance including members of Saccharomycotina and Taphrinomycotina. A comp...

  4. Identification of six loci in which mutations partially restore peroxisome biogenesis and/or alleviate the metabolic defect of pex2 mutants in podospora.

    PubMed Central

    Ruprich-Robert, Gwenaël; Berteaux-Lecellier, Véronique; Zickler, Denise; Panvier-Adoutte, Arlette; Picard, Marguerite

    2002-01-01

    Peroxins (PEX) are proteins required for peroxisome biogenesis. Mutations in PEX genes cause lethal diseases in humans, metabolic defects in yeasts, and developmental disfunctions in plants and filamentous fungi. Here we describe the first large-scale screening for suppressors of a pex mutation. In Podospora anserina, pex2 mutants exhibit a metabolic defect [inability to grow on medium containing oleic acid (OA medium) as sole carbon source] and a developmental defect (inability to differentiate asci in homozygous crosses). Sixty-three mutations able to restore growth of pex2 mutants on OA medium have been analyzed. They fall in six loci (suo1 to suo6) and act as dominant, allele-nonspecific suppressors. Most suo mutations have pleiotropic effects in a pex2(+) background: formation of unripe ascospores (all loci except suo5 and suo6), impaired growth on OA medium (all loci except suo4 and suo6), or sexual defects (suo4). Using immunofluorescence and GFP staining, we show that peroxisome biogenesis is partially restored along with a low level of ascus differentiation in pex2 mutant strains carrying either the suo5 or the suo6 mutations. The data are discussed with respect to beta-oxidation of fatty acids, peroxisome biogenesis, and cell differentiation. PMID:12136013

  5. PHYLOGENETICS OF SACCHAROMYCETALES, THE ASCOMYCETE YEASTS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascomycete yeasts (Phylum Ascomycota: Subphylum Saccharomycotina: Class Saccharomycetes: Order Saccharomycetales) comprise a monophyletic lineage with a single order of about 1000 known species. These yeasts live as saprobes, often in association with plants, animals, and their interfaces. A few s...

  6. Genomic evolution of the ascomycetous yeasts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yeasts are important for industrial and biotechnological processes and show remarkable metabolic and phylogenetic diversity despite morphological similarities. We have sequenced the genomes of 16 ascomycete yeasts of taxonomic and industrial importance including members of Saccharomycotina and Taphr...

  7. Species Diversity of Hypogeous Ascomycetes in Israel

    PubMed Central

    Wasser, Solomon P.

    2010-01-01

    We conducted a species diversity study of the hypogeous Ascomycetes of Israel. The hypogeous Ascomycetes in Israel include members of the families Pyronemataceae, Pezizaceae, and Tuberaceae, which are represented by seven species: Hydnocystis piligera, Terfezia arenaria, T. claveryi, T. oligosperma, Tirmania africana, Tuber asa, and T. nitidum; only T. asa is new to Israeli mycobiota. Synonymy, locations, collection data, general distribution, distribution in Israel, descriptions, a key to identification, illustrations, and taxonomic remarks are provided. PMID:23956647

  8. Species diversity of hypogeous ascomycetes in Israel.

    PubMed

    Barseghyan, Gayane S; Wasser, Solomon P

    2010-09-01

    We conducted a species diversity study of the hypogeous Ascomycetes of Israel. The hypogeous Ascomycetes in Israel include members of the families Pyronemataceae, Pezizaceae, and Tuberaceae, which are represented by seven species: Hydnocystis piligera, Terfezia arenaria, T. claveryi, T. oligosperma, Tirmania africana, Tuber asa, and T. nitidum; only T. asa is new to Israeli mycobiota. Synonymy, locations, collection data, general distribution, distribution in Israel, descriptions, a key to identification, illustrations, and taxonomic remarks are provided. PMID:23956647

  9. Inactivation of the Podospora anserina vegetative incompatibility locus het-c, whose product resembles a glycolipid transfer protein, drastically impairs ascospore production.

    PubMed Central

    Saupe, S; Descamps, C; Turcq, B; Bégueret, J

    1994-01-01

    The het-c locus contains different alleles that elicit nonallelic vegetative incompatibility through specific interactions with alleles of the unlinked loci het-e and het-d. The het-c2 allele has been cloned. It encodes a 208-amino acid polypeptide that is similar to a glycolipid transfer protein purified from pig brain. Disruption of this gene drastically impairs ascospore production in homozygous crosses, and some mutants exhibit abnormal branching of apical hyphae. The protein encoded by het-c2 is essential in the biology of the fungus and may be involved in cell-wall biosynthesis. Images PMID:8016091

  10. IDC1, a pezizomycotina-specific gene that belongs to the PaMpk1 MAP kinase transduction cascade of the filamentous fungus Podospora anserina.

    PubMed

    Jamet-Vierny, Corinne; Debuchy, Robert; Prigent, Magali; Silar, Philippe

    2007-12-01

    Components involved in the activation of the MAPK cascades in filamentous fungi are not well known. Here, we provide evidence that IDC1, a pezizomycotina-specific gene is involved along with the PaNox1 NADPH oxidase in the nuclear localization of the PaMpk1 MAP kinase, a prerequisite for MAPK activity. Mutants of IDC1 display the same phenotypes as mutants in PaNox1 and PaMpk1, i.e., lack of pigment and of aerial hyphae, female sterility, impairment in hyphal interference and inability to develop Crippled Growth cell degeneration. As observed for the PaNox1 mutant, IDC1 mutants are hypostatic to PaMpk1 mutants. IDC1 seems to play a key role in sexual reproduction. Indeed, fertility is diminished in strains with lower level of IDC1. In strains over-expressing IDC1, protoperithecia reach a later stage of development towards perithecia without fertilization; however, upon fertilization maturation of fertile perithecia is diminished and delayed. In addition, heterokaryon construction shows that IDC1 is necessary together with PaNox1 in the perithecial envelope but not in the dikaryon resulting from fertilization. PMID:17517525

  11. The Kinetochore Interaction Network (KIN) of ascomycetes

    PubMed Central

    Freitag, Michael

    2016-01-01

    Chromosome segregation relies on coordinated activity of a large assembly of proteins, the “Kinetochore Interaction Network” (KIN). How conserved the underlying mechanisms driving the epigenetic phenomenon of centromere and kinetochore assembly and maintenance are remains unclear, even though various eukaryotic models have been studied. More than 50 different proteins, many in multiple copies, comprise the KIN or are associated with fungal centromeres and kinetochores. Proteins isolated from immune sera recognized centromeric regions on chromosomes and were thus named centromere proteins (“CENPs”). CENP-A, sometimes called “centromere-specific H3” (CenH3), is incorporated into nucleosomes within or near centromeres. The “constitutive centromere-associated network” (CCAN) assembles on this specialized chromatin, likely based on specific interactions with and requiring presence of CENP-C. The outer kinetochore comprises the Knl1-Mis12-Ndc80 (“KMN”) protein complexes that connect the CCAN to spindles, accomplished by binding and stabilizing microtubules (MTs) and in the process generating load-bearing assemblies for chromatid segregation. In most fungi the Dam1/DASH complex connects the KMN complexes to MTs. Fungi present a rich resource to investigate mechanistic commonalities but also differences in kinetochore architecture. While ascomycetes have sets of CCAN and KMN proteins that are conserved with those of either budding yeast or metazoans, searching other major branches of the fungal kingdom revealed that CCAN proteins are poorly conserved at the primary sequence level. Several conserved binding motifs or domains within KMN complexes have been described recently, and these features of ascomycete KIN proteins are shared with most metazoan proteins. In addition, several ascomycete-specific domains have been identified here. PMID:26908646

  12. The kinetochore interaction network (KIN) of ascomycetes.

    PubMed

    Freitag, Michael

    2016-05-01

    Chromosome segregation relies on coordinated activity of a large assembly of proteins, the kinetochore interaction network (KIN). How conserved the underlying mechanisms driving the epigenetic phenomenon of centromere and kinetochore assembly and maintenance are remains unclear, even though various eukaryotic models have been studied. More than 50 different proteins, many in multiple copies, comprise the KIN or are associated with fungal centromeres and kinetochores. Proteins isolated from immune sera recognized centromeric regions on chromosomes and thus were named centromere proteins (CENPs). CENP-A, sometimes called centromere-specific H3 (CenH3), is incorporated into nucleosomes within or near centromeres. The constitutive centromere-associated network (CCAN) assembles on this specialized chromatin, likely based on specific interactions with and requiring presence of CENP-C. The outer kinetochore comprises the Knl1-Mis12-Ndc80 (KMN) protein complexes that connect CCAN to spindles, accomplished by binding and stabilizing microtubules (MTs) and in the process generating load-bearing assemblies for chromatid segregation. In most fungi the Dam1/DASH complex connects the KMN complexes to MTs. Fungi present a rich resource to investigate mechanistic commonalities but also differences in kinetochore architecture. While ascomycetes have sets of CCAN and KMN proteins that are conserved with those of budding yeast or metazoans, searching other major branches of the fungal kingdom revealed that CCAN proteins are poorly conserved at the primary sequence level. Several conserved binding motifs or domains within KMN complexes have been described recently, and these features of ascomycete KIN proteins are shared with most metazoan proteins. In addition, several ascomycete-specific domains have been identified here. PMID:26908646

  13. Acylated proteins in Borrelia hermsii, Borrelia parkeri, Borrelia anserina, and Borrelia coriaceae.

    PubMed Central

    Sambri, V; Stefanelli, C; Rossoni, C; La Placa, M; Cevenini, R

    1993-01-01

    Borrelia hermsii, Borrelia parkeri, Borrelia anserina, and Borrelia coriaceae produced several lipoproteins identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of bacteria grown in [3H]palmitate. Five major acylated proteins were demonstrated by sequential alkaline and acid hydrolysis. High-pressure liquid chromatography of isolated proteins confirmed that covalently bound radioactivity was represented by fatty acids. Images PMID:8285697

  14. Prevalence of transcription factors in ascomycete and basidiomycete fungi

    PubMed Central

    2014-01-01

    Background Gene regulation underlies fungal physiology and therefore is a major factor in fungal biodiversity. Analysis of genome sequences has revealed a large number of putative transcription factors in most fungal genomes. The presence of fungal orthologs for individual regulators has been analysed and appears to be highly variable with some regulators widely conserved and others showing narrow distribution. Although genome-scale transcription factor surveys have been performed before, no global study into the prevalence of specific regulators across the fungal kingdom has been presented. Results In this study we have analysed the number of members for 37 regulator classes in 77 ascomycete and 31 basidiomycete fungal genomes and revealed significant differences between ascomycetes and basidiomycetes. In addition, we determined the presence of 64 regulators characterised in ascomycetes across these 108 genomes. This demonstrated that overall the highest presence of orthologs is in the filamentous ascomycetes. A significant number of regulators lacked orthologs in the ascomycete yeasts and the basidiomycetes. Conversely, of seven basidiomycete regulators included in the study, only one had orthologs in ascomycetes. Conclusions This study demonstrates a significant difference in the regulatory repertoire of ascomycete and basidiomycete fungi, at the level of both regulator class and individual regulator. This suggests that the current regulatory systems of these fungi have been mainly developed after the two phyla diverged. Most regulators detected in both phyla are involved in central functions of fungal physiology and therefore were likely already present in the ancestor of the two phyla. PMID:24650355

  15. Genomic Evolution of the Ascomycete Yeasts

    SciTech Connect

    Riley, Robert; Haridas, Sajeet; Salamov, Asaf; Boundy-Mills, Kyria; Goker, Markus; Hittinger, Chris; Klenk, Hans-Peter; Lopes, Mariana; Meir-Kolthoff, Jan P.; Rokas, Antonis; Rosa, Carlos; Scheuner, Carmen; Soares, Marco; Stielow, Benjamin; Wisecaver, Jennifer H.; Wolfe, Ken; Blackwell, Meredith; Kurtzman, Cletus; Grigoriev, Igor; Jeffries, Thomas

    2015-03-16

    Yeasts are important for industrial and biotechnological processes and show remarkable metabolic and phylogenetic diversity despite morphological similarities. We have sequenced the genomes of 16 ascomycete yeasts of taxonomic and industrial importance including members of Saccharomycotina and Taphrinomycotina. Phylogenetic analysis of these and previously published yeast genomes helped resolve the placement of species including Saitoella complicata, Babjeviella inositovora, Hyphopichia burtonii, and Metschnikowia bicuspidata. Moreover, we find that alternative nuclear codon usage, where CUG encodes serine instead of leucine, are monophyletic within the Saccharomycotina. Most of the yeasts have compact genomes with a large fraction of single exon genes, and a tendency towards more introns in early-diverging species. Analysis of enzyme phylogeny gives insights into the evolution of metabolic capabilities such as methanol utilization and assimilation of alternative carbon sources.

  16. Discussion of teleomorphic and anamorphic Ascomycetous yeasts and yeast-like taxa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The relationship of ascomycetous yeasts with other members of the ascomycete fungi (Ascomycota) has been controversial for over 100 years. Because yeasts are morphologically simple, it was proposed that they represent primitive forms of ascomycetes (e.g., Guilliermond 1912). Alternatively, the ide...

  17. Interactive effects of pollination and heavy metals on resource allocation in Potentilla anserina L.

    SciTech Connect

    Saikkonen, K. |; Koivunen, S.; Vuorisalo, T.; Mutikainen, P. |

    1998-07-01

    The authors studied resource allocation between sexual reproduction and clonal propagation in a perennial stoloniferous clonal plant, Potentilla anserina, an obligate outcrosser. They manipulated reproductive effort of Potentilla anserina either by hand-pollinating all flowers or by preventing pollination. To test the effect of resource-limiting conditions on resource allocation and reproductive output, the authors used a control and two levels of heavy metals (copper and nickel) to limit plant growth. The experiment was conducted as a 2 {times} 3 factorial design to reveal possible interactions between reproductive manipulation and resource limitation. Heavy metals decreased the total biomass of the plants and number of flowers and ramets produced. Only 50% of the plants grown with the higher level of heavy metals produced flowers. Pollination treatment interacted significantly with the heavy-metal treatment. In the metal control and lower heavy-metal treatment, there were no significant differences in total vegetative biomass between the two pollination treatments. Costs of reproduction in terms of subsequent flowering in the later season appeared to be clear, because the number of flowers per whole plant was lower if the plants were hand-pollinated and because the proportion of flowering ramets decreased due to hand-pollination. However, flowering may also be partly hormonally controlled. In contrast, hand-pollinated plants exposed to high concentrations of heavy metals tended to have greater biomass of vegetative plant structures and higher number of flowers compared to nonpollinated plants.

  18. NOTES ON ASCOMYCETE SYSTEMATICS NOS 3303-3579

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The series "Notes on ascomycete systematics" has been published in Systema Ascomycetum (Eriksson & Hawksworth 1986-1998) and since 1999 in Myconet in an electronic version on the Internet (http://www.umu.se/myconet/notes.html) and as hard copies once or twice a year in a journal with the same name (...

  19. ASCOMYCETOUS MITOSIS IN BASIDIOMYCETOUS YEASTS: ITS EVOLUTIONARY IMPLICATIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In budding cells of ascomycetous yeasts, mitosis occurs in the parent, while in basidiomyceteous yeasts it occurs in the bud. However, in the basidiomycete Agaricostilbum pulcherrimum mitosis occurs in the parent and parent-bud junction. To test whether A. pulcherrimum has a novel mitotic pattern, i...

  20. Intronic GIY-YIG endonuclease gene in the mitochondrial genome of Podospora curvicolla: evidence for mobility

    PubMed Central

    Saguez, Cyril; Lecellier, Gaël; Koll, France

    2000-01-01

    Endonuclease genes encoded in invasive introns are themselves supposed to be mobile elements which, during evolution, have colonized pre-existing introns converting them into invasive elements. This hypothesis is supported by numerous data concerning the LAGLI-DADG subclass of intronic endonucleases. Less is known about the GIY-YIG ORFs which constitute another family of endonucleases. In this paper we describe the presence of one optional GIY-YIG ORF in the second intron of the mitochondrial cytochrome b gene in the fungus Podospora curvicolla. We show that this GIY-YIG ORF is efficiently transferred from an ORF-containing intron to an ORF-less allele. We also show that the products of both the GIY-YIG ORF and the non-canonical LAGLI-DADG-GIY-YIG ORF, which is generated by its integration, have endonuclease activities which recognize and cut the insertion site of the optional sequence. This constitutes the first direct evidence for potential mobility of an intronic GIY-YIG endonuclease. We discuss the role that such a mobile sequence could have played during evolution. PMID:10684923

  1. Specific Antibodies Reactive with the 22-Kilodalton Major Outer Surface Protein of Borrelia anserina Ni-NL Protect Chicks from Infection

    PubMed Central

    Sambri, Vittorio; Marangoni, Antonella; Olmo, Andrea; Storni, Elisa; Montagnani, Marco; Fabbi, Massimo; Cevenini, Roberto

    1999-01-01

    An outer surface lipoprotein of 22 kDa was identified in the avian pathogen Borrelia anserina Ni-NL by using antibody preparations reactive with bacterial surface-exposed proteins. Amino acid sequence analysis of the 22-kDa protein demonstrated 90% identity with VmpA of B. turicatae, suggesting that the protein belongs to the family of 20-kDa outer surface proteins of the genus Borrelia. All of the 60 chicks intramuscularly treated with antibodies specifically reacting with the 22-kDa protein and infected with strain Ni-NL were completely protected from infection, since no spirochetemia was detected, and from death. Control chicks were treated with immune sera raised against apathogenic strain B. anserina Es, which expresses a prominent 20-kDa polypeptide that is also a member of the Vmp family but does not cross-react immunologically with the 22-kDa protein of the Ni-NL strain. These animals, infected with B. anserina Ni-NL, showed a high degree of spirochetemia 10 days after infection, and all died between 14 and 21 days after infection. The results showed that the 22-kDa surface protein of B. anserina Ni-NL is a determinant of the pathogenic potential of the strain and also confirmed that only strain-specific antibodies are protective against B. anserina infection. PMID:10225933

  2. The Chemical Diversity of the Ascomycete Fungus Paecilomyces variotii.

    PubMed

    Mioso, Roberto; Toledo Marante, Francisco Javier; Herrera Bravo de Laguna, Irma

    2015-10-01

    Paecilomyces variotii isolated from a broad range of habitats drives the diversification of new high-value-added secondary metabolites that could potentially play an important role in human and animal health. These metabolites include the anhydride metabolite of the nonadride family, as well as the following compounds: naphthopyranone metabolites, sphingofungins, eicosenoic acids, new branched fatty acids, ascofuranone, polyketides, an anacardic acid analogue, straight-chain peptides, and volatile compounds. These natural products show that P. variotii can provide leading compounds for new drug discoveries, which may include herbicide agents, some of which are important in the agrochemical market. Finally, this review outlines recent developments, trends, and prospects for the chemistry of this ascomycete. PMID:26288080

  3. Genomics of alternative sulfur utilization in ascomycetous yeasts.

    PubMed

    Linder, Tomas

    2012-10-01

    Thirteen ascomycetous yeast strains with sequenced genomes were assayed for their ability to grow on chemically defined medium with 16 different sulfur compounds as the only significant source of sulfur. These compounds included sulfoxides, sulfones, sulfonates, sulfamates and sulfate esters. Broad utilization of alternative sulfur sources was observed in Komagataella pastoris (syn. Pichia pastoris), Lodderomyces elongisporus, Millerozyma farinosa (syn. Pichia sorbitophila), Pachysolen tannophilus, Scheffersomyces stipitis (syn. Pichia stipitis), Spathaspora passalidarum, Yamadazyma tenuis (syn. Candida tenuis) and Yarrowia lipolytica. Kluyveromyces lactis, Saccharomyces cerevisiae and Zygosaccharomyces rouxii were mainly able to utilize sulfonates and sulfate esters, while Lachancea thermotolerans and Schizosaccharomyces pombe were limited to aromatic sulfate esters. Genome analysis identified several candidate genes with bacterial homologues that had been previously shown to be involved in the utilization of alternative sulfur sources. Analysis of candidate gene promoter sequences revealed a significant overrepresentation of DNA motifs that have been shown to regulate sulfur metabolism in Sacc. cerevisiae. PMID:22790398

  4. Identification of Oxaphenalenone Ketals from the Ascomycete Fungus Neonectria sp.

    PubMed

    Ren, Jinwei; Niu, Shubing; Li, Li; Geng, Zhufeng; Liu, Xingzhong; Che, Yongsheng

    2015-06-26

    Neonectrolides B-E (4-7), four new oxaphenalenone ketals incorporating the new furo[2,3-b]isochromeno[3,4,5-def]chromen-11(6aH)-one skeleton, were isolated from the fermentation extract of the ascomycete fungus Neonectria sp. in an in-depth investigation guided by HPLC fingerprint and a cytotoxicity assay. The previously identified oxaphenalenone spiroketal neonectrolide A (1) and its putative biosynthetic precursors (2 and 3) were also reisolated in the current work. The structures of 4-7 were primarily elucidated by interpretation of NMR spectroscopic data, and the absolute configurations were deduced by electronic circular dichroism calculations. Compound 6 showed cytotoxic effects against four of the six human tumor cell lines tested. Biosynthetically, compounds 4-7 could be derived via the Diels-Alder reaction cascades starting from derivatives of the co-isolated metabolites 2 and 3. PMID:25978132

  5. Phytochemical composition of Potentilla anserina L. analyzed by an integrative GC-MS and LC-MS metabolomics platform.

    PubMed

    Mari, Angela; Lyon, David; Fragner, Lena; Montoro, Paola; Piacente, Sonia; Wienkoop, Stefanie; Egelhofer, Volker; Weckwerth, Wolfram

    2013-06-01

    Potentilla anserina L. (Rosaceae) is known for its beneficial effects of prevention of pre-menstrual syndrome (PMS). For this reason P. anserina is processed into many food supplements and pharmaceutical preparations. Here we analyzed hydroalcoholic reference extracts and compared them with various extracts of different pharmacies using an integrative metabolomics platform comprising GC-MS and LC-MS analysis and software toolboxes for data alignment (MetMAX Beta 1.0) and multivariate statistical analysis (COVAIN 1.0). Multivariate statistics of the integrated GC-MS and LC-MS data showed strong differences between the different plant extract formulations. Different groups of compounds such as chlorogenic acid, kaempferol 3-O-rutinoside, acacetin 7-O-rutinoside, and genistein were reported for the first time in this species. The typical fragmentation pathway of the isoflavone genistein confirmed the identification of this active compound that was present with different abundances in all the extracts analyzed. As a result we have revealed that different extraction procedures from different vendors produce different chemical compositions, e.g. different genistein concentrations. Consequently, the treatment may have different effects. The integrative metabolomics platform provides the highest resolution of the phytochemical composition and a mean to define subtle differences in plant extract formulations. PMID:23678344

  6. Genomic exploration of the hemiascomycetous yeasts: 19. Ascomycetes-specific genes.

    PubMed

    Malpertuy, A; Tekaia, F; Casarégola, S; Aigle, M; Artiguenave, F; Blandin, G; Bolotin-Fukuhara, M; Bon, E; Brottier, P; de Montigny, J; Durrens, P; Gaillardin, C; Lépingle, A; Llorente, B; Neuvéglise, C; Ozier-Kalogeropoulos, O; Potier, S; Saurin, W; Toffano-Nioche, C; Wésolowski-Louvel, M; Wincker, P; Weissenbach, J; Souciet, J; Dujon, B

    2000-12-22

    Comparisons of the 6213 predicted Saccharomyces cerevisiae open reading frame (ORF) products with sequences from organisms of other biological phyla differentiate genes commonly conserved in evolution from 'maverick' genes which have no homologue in phyla other than the Ascomycetes. We show that a majority of the 'maverick' genes have homologues among other yeast species and thus define a set of 1892 genes that, from sequence comparisons, appear 'Ascomycetes-specific'. We estimate, retrospectively, that the S. cerevisiae genome contains 5651 actual protein-coding genes, 50 of which were identified for the first time in this work, and that the present public databases contain 612 predicted ORFs that are not real genes. Interestingly, the sequences of the 'Ascomycetes-specific' genes tend to diverge more rapidly in evolution than that of other genes. Half of the 'Ascomycetes-specific' genes are functionally characterized in S. cerevisiae, and a few functional categories are over-represented in them. PMID:11152894

  7. Common amino acid domain among endopolygalacturonases of ascomycete fungi.

    PubMed Central

    Keon, J P; Waksman, G

    1990-01-01

    The endopolygalacturonase (EC 3.2.1.15) enzymes produced in vitro by three ascomycete fungi, Aspergillus niger, Sclerotinia sclerotiorum, and Colletotrichum lindemuthianum were studied by using thin-layer isoelectric focusing and activity stain overlay techniques. The polygalacturonases from A. niger and S. sclerotiorum consisted of numerous isoforms, whereas the endopolygalacturonase from C. lindemuthianum consisted of a single protein species. The most abundant endopolygalacturonase isoform produced by each of these organisms was purified and characterized. Biochemical parameters, including molecular weight, isoelectric point, kinetic parameters, temperature and pH optima, and thermal stability, were determined. Considerable differences in physical and chemical properties were demonstrated among these fungal polygalacturonases. Antibodies raised against individual proteins exhibited little cross-reaction, suggesting that these enzymes differ structurally as well as biochemically. In contrast, the analysis of the N-terminal amino acid sequences of the three proteins showed extensive homology, particularly in a region labeled domain 1 in which 84% of the amino acids were conserved. Images PMID:2403258

  8. A novel class of peptide pheromone precursors in ascomycetous fungi

    PubMed Central

    Schmoll, Monika; Seibel, Christian; Tisch, Doris; Dorrer, Marcel; Kubicek, Christian P

    2010-01-01

    Recently, sexual development in the heterothallic ascomycete Trichoderma reesei (anamorph of Hypocrea jecorina) has been achieved and thus initiated attempts to elucidate regulation and determinants of this process. While the α-type pheromone of this fungus fits the consensus known from other fungi, the assumed a-type peptide pheromone precursor shows remarkably unusual characteristics: it comprises three copies of the motif (LI)GC(TS)VM thus constituting a CAAX domain at the C-terminus and two Kex2-protease sites. This structure shares characteristics of both a- and α-type peptide pheromone precursors. Presence of hybrid-type peptide pheromone precursor 1 (hpp1) is essential for male fertility, thus indicating its functionality as a peptide pheromone precursor, while its phosphorylation site is not relevant for this process. However, sexual development in a female fertile background is not perturbed in the absence of hpp1, which rules out a higher order function in this process. Open reading frames encoding proteins with similar characteristics to HPP1 were also found in Fusarium spp., of which Fusarium solani still retains a putative a-factor-like protein, but so far in no other fungal genome available. We therefore propose the novel class of h-type (hybrid) peptide pheromone precursors with H. jecorina HPP1 as the first member of this class. PMID:20735770

  9. Lipids stimulate spore germination in the entomopathogenic ascomycete Ascosphaera aggregata.

    PubMed

    James, R R; Buckner, J S

    2004-10-01

    The alfalfa leafcutting bee (Megachile rotundata) is solitary and managed on a large scale for pollination of alfalfa seed crops. The bees nest in holes drilled in wood or polystyrene blocks, and their larvae are highly prone to a fungal disease called chalkbrood. The most prevalent form of chalkbrood is caused by Ascosphaera aggregata, but this ascomycete is difficult to culture. Hyphae will grow on standard fungal media, but spore germination is difficult to achieve and highly variable. We found that germination can be enhanced with oils. Lipids derived from plants and bee larvae increased germination from 50% (without oil) to 75-85% (with oil). Percent germination was significantly greater in the presence of lipids but germination was not significantly different when different oils, including mineral oil, were used. A. aggregata spores oriented along the oil-aqueous interface in the broth in a polar fashion, with swelling and germ tube formation always occurring into the aqueous portion of the broth. The other half of the spore tended to attach to a lipid droplet, where it remained, without swelling, during germ tube formation. The physical attachment of spores to the oil-aqueous interface is what most probably stimulates spore germination, as opposed to some nutritional stimulation. However, further research is needed to determine if and where the spores encounter such an interface when germinating in the host gut, where germination normally occurs. PMID:15645171

  10. Mycelial pellet formation by edible ascomycete filamentous fungi, Neurospora intermedia.

    PubMed

    Nair, Ramkumar B; Lennartsson, Patrik R; Taherzadeh, Mohammad J

    2016-12-01

    Pellet formation of filamentous fungi in submerged culture is an imperative topic of fermentation research. In this study, we report for the first time the growth of filamentous ascomycete fungus, Neurospora intermedia in its mycelial pellet form. In submerged culture, the growth morphology of the fungus was successfully manipulated into growing as pellets by modifying various cultivation conditions. Factors such as pH (2.0-10.0), agitation rate (100-150 rpm), carbon source (glucose, arabinose, sucrose, and galactose), the presence of additive agents (glycerol and calcium chloride) and trace metals were investigated for their effect on the pellet formation. Of the various factors screened, uniform pellets were formed only at pH range 3.0-4.0, signifying it as the most influential factor for N. intermedia pellet formation. The average pellet size ranged from 2.38 ± 0.12 to 2.86 ± 0.38 mm. The pellet formation remained unaffected by the inoculum type used and its size showed an inverse correlation with the agitation rate of the culture. Efficient glucose utilization was observed with fungal pellets, as opposed to the freely suspended mycelium, proving its viability for fast-fermentation processes. Scale up of the pelletization process was also carried out in bench-scale airlift and bubble column reactors (4.5 L). PMID:27103628

  11. Phylogenetic studies of four Anser cygnoides (Anserini: Anserinae) in Hunan province of China based on complete mitochondrial DNA sequences.

    PubMed

    Dai, Qiu-Zhong; Lin, Qian; Jiang, Gui-Tao

    2016-07-01

    In this study, we cloned and sequenced the complete mitochondrial DNAs of Chinese goose, Anser cygnoides populations from three different areas of Hunan province in China. The Anser cygnoides breed Wugangtong white goose (WGTW) sample and Wugangtong grey goose sample (WGTG) were taken from the Wugang county of Shaoyang city, the Anser cygnoides breed Xupu goose (XP) sample was taken from the Xupu county of Huaihua city, and the Anser cygnoides breed Yanling white goose (YLW) sample was taken from the Yanling county of Zhuzhou city. The organization of the four Anser cygnoides breeds mitochondrial genomes was similar. Phylogenetic analyses using N-J computational algorithms showed that the analyzed species are divided into four major clades: Anatinae, Anserinae, Dendrocygninae and Anseranatidae. It was noted that Wugangtong white goose, Yanling white goose and Xupu goose have highly similar phylogenetic relationship. PMID:26057006

  12. Purification and properties of a lectin from ascomycete mushroom, Ciborinia camelliae.

    PubMed

    Otta, Yumi; Amano, Koh; Nishiyama, Kano; Ando, Akikazu; Ogawa, Shigeru; Nagata, Yoshiho

    2002-05-01

    A lectin was isolated from an ascomycete mushroom, Ciborinia camelliae which was specific to N-acetyl-D-galactosamine. On SDS-polyacrylamide gel electrophoresis; this lectin gave a single band of approximately 17-kDa in the presence of 2-mercaptoethanol, but formed dimers, trimers and tetramers in its absence. Amino acid analysis revealed the lectin contained two cysteines and no methionine. The N-terminal sequence was determined up to residue 21, and no homologous proteins including other ascomycete lectins were found. PMID:12009312

  13. Phylogeny of the ascomycetous yeasts and the renaming of Pichia anomala to Wickerhamomyces anomalus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pichia anomala was reclassified as Wickerhamomyces anomalus following multigene phylogenetic analysis. In this review, the phylogeny of the ascomycetous yeasts is discussed, with emphasis on the genus Pichia. The genus, as defined from phenotype, had nearly 100 assigned species, but the number of ...

  14. Two ascomycete classes based on fruiting-body characters and ribosomal DNA sequence.

    PubMed

    Berbee, M L; Taylor, J W

    1992-03-01

    Traditional fruiting body-based classification of ascomycetes has been under attack for 2 decades. Fruiting-body types can converge, and few researchers now assume that either the closed fruiting bodies (cleistothecia) characterizing the class Plectomycetes or the flask-shaped fruiting bodies (perithecia) characterizing the class Pyrenomycetes are stable, unifying characters. Unless we identify characters uniting major ascomycete groups, orders of ascomycetes remain narrowly defined, and supraordinal classification is impossible. We sequenced both strands of 18s rDNA from nine ascomycete fungi, adding three sequences from GenBank into our analysis. The phylogeny, inferred from 162 informative sites in 1,700 bp of DNA sequence data and using yeast as an outgroup, divided the fungi into two groups correlating well both with fruiting-body type and with the traditional classes Plectomycetes and Pyrenomycetes. Each group received strong statistical support. Genera producing cleistothecia, such as Talaromyces (with a Penicillium asexual state) and the human pathogen Ajellomyces capsulatus (causing histoplasmosis), fall within the plectomycete group. Plectomycetes also includes Eremascus albus and the bee pathogen Ascosphaera apis, although both lack typical fruiting bodies. The Dutch elm disease fungus groups with pyrenomycetes such as Neurospora, in spite of its confusing mixture of class-level characters. PMID:1560763

  15. Morphological and molecular characteristics of a poorly known marine ascomycete, Manglicola guatemalensis (Jahnulales: Pezizomycotina; Dothideomycetes, Incertae sedis): new lineage of marine ascomycetes.

    PubMed

    Suetrong, Satinee; Sakayaroj, Jariya; Phongpaichit, Souwalak; Jones, E B Gareth

    2010-01-01

    The poorly known marine ascomycete Manglicola guatemalensis from Trang and Trat provinces, Thailand, were collected in 2005 and 2006 on the brackish water palm Nypa fruticans. This fungus is known only from two previous collections. This paper reports on the morphological characteristics and molecular phylogeny of this unique marine bitunicate ascomycete. Manglicola guatemalensis has large clavate to obtusely fusiform ascomata, wide ostioles, bitunicate asci, cylindrical, thick-walled, unequally one-septate ascospores, constricted at the septum, apical cell larger, chestnut-brown and a smaller light brown basal cell. Ascospores germinate readily, always from the basal cell. Four isolates from different locations were selected for the phylogenetic study. Regions of the rDNA gene, including SSU and LSU, were sequenced and combined. Molecular data places M. guatemalensis in the Jahnulales with high bootstrap support; all strains are monophyletic. In the combined SSU and LSU analyses the Jahnulales comprises four subclades, A, B, C and D. Subclade A comprises Jahnula species and two anamorphic fungi, Brachiosphaera tropicalis and Xylomyces chlamydosporus. In subclade B Manglicola strains form a sister group of the Aliquandostipite species Aliquandostipite crystallinus, A. khaoyaiensis, Jahnula siamensiae and Patescospora separans. Subclade C comprises Jahnula species, Jahnula aquatica, J. granulosa and J. rostrata, while Megalohypha aqua-dulces constitutes subclade D. Therefore Manglicola forms another lineage of marine fungi. PMID:20120232

  16. Low pH dye decolorization with ascomycete Lamprospora wrightii laccase.

    PubMed

    Mueangtoom, Kitti; Kittl, Roman; Mann, Oliver; Haltrich, Dietmar; Ludwig, Roland

    2010-08-01

    In a screening of saprotrophic, ectomycorrhizal and plant pathogen ascomycetes a frequent occurrence of laccase was observed. Lamprospora wrightii, the best producing organism, was chosen to elucidate the properties of a laccase from a moss-associated, saprotrophic ascomycete. The expression of laccase by this bryophilic fungus could be increased by the addition of tomato juice or copper sulfate to the medium. The obtained volumetric activity after optimization was 420 U/mL in either shaking flask or bioreactor-based cultivations. The purified laccase has a molecular mass of 68 kDa and an isoelectric point of 3.4. Although of ascomycete origin, its catalytic properties are similar to typical basidiomycte laccases, and an excellent activity and stability was observed at low pH, which makes it suitable for bioremediation in acidic environments. As an example, the decolorization reactions of azo-, anthraquinone-, trimethylmethane- and indigoid dyes at pH 3.0 and 5.0 were investigated. All ten selected dyes were decolorized, five of them very efficiently. Depending on the dye, the decolorization was found to be a combination of two reactions, degradation of the chromophore and formation of polymerized products, which contributed to the overall process in a dye-specific pattern. PMID:20652905

  17. A search for the phylogenetic relationship of the ascomycete Rhizoctonia leguminicola using genetic analysis.

    PubMed

    Alhawatema, Mohammad S; Sanogo, Soum; Baucom, Deana L; Creamer, Rebecca

    2015-06-01

    Rhizoctonia leguminicola, which causes fungal blackpatch disease of legumes and other plants, produces slaframine and swainsonine that are largely responsible for causing salivation, lacrimation, frequent urination, and diarrhea in grazing animals including cattle, sheep, and horses. The original identification of R. leguminicola was based only on morphological characters of the fungal mycelia in cultures because of the lack of fungal genetic markers. Recent investigations suggested that R. leguminicola does not belong to genus Rhizoctonia and is instead a member of the ascomycetes, necessitating an accurate reclassification. The objective of this study was to use both genetic and morphological characters of R. leguminicola to find taxonomic placement of this pathogen within ascomycetes. Internal transcribed spacer region (ITS) and glyceraldehyde-3-phosphate dehydrogenase (gpd) encoding gene were amplified from R. leguminicola isolates by PCR using universal primers and sequencing. Rhizoctonia leguminicola ITS and gpd sequences were aligned with other fungal sequences of close relatives, and phylogenetic trees were constructed using neighbor-joining and parsimony analyses. Rhizoctonia leguminicola isolates were clustered within a clade that contains several genera of ascomycetes belonging to the class dothideomycetes. We suggest that the fungus is misidentified in the genus Rhizoctonia and propose its reclassification in a new genus within the phylum Ascomycota. PMID:25585493

  18. Self/nonself recognition in Tuber melanosporum is not mediated by a heterokaryon incompatibility system.

    PubMed

    Iotti, Mirco; Rubini, Andrea; Tisserant, Emilie; Kholer, Annegret; Paolocci, Francesco; Zambonelli, Alessandra

    2012-02-01

    Vegetative incompatibility is a widespread phenomenon in filamentous ascomycetes, which limits formation of viable heterokaryons. Whether this phenomenon plays a role in maintaining the homokaryotic state of the hyphae during the vegetative growth of Tuber spp. Gene expression, polymorphism analysis as well as targeted in vitro experiments allowed us to test whether a heterokaryon incompatibility (HI) system operates in Tuber melanosporum. HI is controlled by different genetic systems, often involving HET domain genes and their partners whose interaction can trigger a cell death reaction. Putative homologues to HI-related genes previously characterized in Neurospora crassa and Podospora anserina were identified in the T. melanosporum genome. However, only two HET domain genes were found. In many other ascomycetes HET domains have been found within different genes including some members of the NWD (NACHT and WD-repeat associated domains) gene family of P. anserina. More than 50 NWD homologues were found in T. melanosporum but none of these contain a HET domain. All these T. melanosporum paralogs showed a conserved gene organization similar to the microexon genes only recently characterized in Schistosoma mansoni. Expression data of the annotated HI-like genes along with low allelic polymorphism suggest that they have cellular functions unrelated to HI. Moreover, morphological analyses did not provide evidence for HI reactions between pairs of genetically different T. melanosporum strains. Thus, the maintenance of the genetic integrity during the vegetative growth of this species likely depends on mechanisms that act before hyphal fusion. PMID:22289772

  19. Evolution of Multicopper Oxidase Genes in Coprophilous and Non-Coprophilous Members of the Order Sordariales

    PubMed Central

    Pggeler, Stefanie

    2011-01-01

    Multicopper oxidases (MCO) catalyze the biological oxidation of various aromatic substrates and have been identified in plants, insects, bacteria, and wood rotting fungi. In nature, they are involved in biodegradation of biopolymers such as lignin and humic compounds, but have also been tested for various industrial applications. In fungi, MCOs have been shown to play important roles during their life cycles, such as in fruiting body formation, pigment formation and pathogenicity. Coprophilous fungi, which grow on the dung of herbivores, appear to encode an unexpectedly high number of enzymes capable of at least partly degrading lignin. This study compared the MCO-coding capacity of the coprophilous filamentous ascomycetes Podospora anserina and Sordaria macrospora with closely related non-coprophilous members of the order Sordariales. An increase of MCO genes in coprophilic members of the Sordariales most probably occurred by gene duplication and horizontal gene transfer events. PMID:21966247

  20. Role of Reactive Intermediates in Manganese Oxide Formation By Filamentous Ascomycete Fungi

    NASA Astrophysics Data System (ADS)

    Zeiner, C. A.; Anderton, C.; Wu, S.; Purvine, S.; Zink, E.; Paša-Tolić, L.; Santelli, C. M.; Hansel, C. M.

    2014-12-01

    Biogenic manganese (Mn) oxide minerals are ubiquitous in the environment, and their high reactivity can profoundly impact the fate of contaminants and cycling of carbon and nutrients. In contrast to bacteria, the pathways utilized by fungi to oxidize Mn(II) to Mn(III,IV) oxides remain largely unknown. Here, we explore the mechanisms of Mn(II) oxidation by a phylogenetically diverse group of filamentous Ascomycete fungi using a combination of chemical assays and bulk and spatially-resolved mass spectrometry. We show that the mechanisms of Mn(II) oxidation vary with fungal species, over time during secretome compositional changes, and in the presence of other fungi. Specifically, our work implicates a dynamic transition in Mn(II) oxidation pathways that varies between species. In particular, while reactive oxygen species (ROS) produced via transmembrane NADPH oxidases are involved in initial oxidation, over time, secreted enzymes become important Mn(II) oxidation mediators for some species. In addition, the overall secretome oxidation capacity varies with time and fungal species. Secretome analysis reveals a surprising absence of enzymes currently considered to be Mn(II)-oxidizing enzymes in these organisms, and instead highlights a wide variety of redox-active enzymes. Furthermore, we implicate fungal cell defense mechanisms in the formation of distinct Mn oxide patterns when fungi are grown in head-to-head competition. The identification and regulation of these secreted enzymes are under current investigation within the bulk secretome and within the interaction zone of structured fungal communities. Overall, our findings illustrate that Ascomycete Mn(II) oxidation mechanisms are highly variable and are dictated by complex environmental and ecological interactions. Future work will explore the connection between Ascomycete Mn(II) oxidation and the ability to degrade cellulose, a key carbon reservoir for biofuel production.

  1. The Potential for pathogenicity was present in the ancestor of the Ascomycete subphylum Pezizomycotina

    PubMed Central

    2010-01-01

    Background Previous studies in Ascomycetes have shown that the function of gene families of which the size is considerably larger in extant pathogens than in non-pathogens could be related to pathogenicity traits. However, by only comparing gene inventories in extant species, no insights can be gained into the evolutionary process that gave rise to these larger family sizes in pathogens. Moreover, most studies which consider gene families in extant species only tend to explain observed differences in gene family sizes by gains rather than by losses, hereby largely underestimating the impact of gene loss during genome evolution. Results In our study we used a selection of recently published genomes of Ascomycetes to analyze how gene family gains, duplications and losses have affected the origin of pathogenic traits. By analyzing the evolutionary history of gene families we found that most gene families with an enlarged size in pathogens were present in an ancestor common to both pathogens and non-pathogens. The majority of these families were selectively maintained in pathogenic lineages, but disappeared in non-pathogens. Non-pathogen-specific losses largely outnumbered pathogen-specific losses. Conclusions We conclude that most of the proteins for pathogenicity were already present in the ancestor of the Ascomycete lineages we used in our study. Species that did not develop pathogenicity seemed to have reduced their genetic complexity compared to their ancestors. We further show that expansion of gained or already existing families in a species-specific way is important to fine-tune the specificities of the pathogenic host-fungus interaction. PMID:20964831

  2. SHARED ITS DNA SUBSTITUTIONS IN ISOLATES OF OPPOSITE MATING TYPE REVEAL A RECOMBIING HISTORY FOR THREE PRESUMED ASEXUAL SPECIES IN THE FILAMENTOUS ASCOMYCETE GENUS ALTERNARIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    About 15,000 species of ascomycete fungi lack a known sexual state. For fungi with asexual states in the form genera Embellisia, Ulocladium and Alternaria, six species have known sexual states but more than 50 species do not. In sexual filamentous ascomycetes, opposite mating type information at t...

  3. Conservation and evolution of cis-regulatory systems in ascomycete fungi

    SciTech Connect

    Gasch, Audrey P.; Moses, Alan M.; Chiang, Derek Y.; Fraser, Hunter B.; Berardini, Mark; Eisen, Michael B.

    2004-03-15

    Relatively little is known about the mechanisms through which gene expression regulation evolves. To investigate this, we systematically explored the conservation of regulatory networks in fungi by examining the cis-regulatory elements that govern the expression of coregulated genes. We first identified groups of coregulated Saccharomyces cerevisiae genes enriched for genes with known upstream or downstream cis-regulatory sequences. Reasoning that many of these gene groups are coregulated in related species as well, we performed similar analyses on orthologs of coregulated S. cerevisiae genes in 13 other ascomycete species. We find that many species-specific gene groups are enriched for the same flanking regulatory sequences as those found in the orthologous gene groups from S. cerevisiae, indicating that those regulatory systems have been conserved in multiple ascomycete species. In addition to these clear cases of regulatory conservation, we find examples of cis-element evolution that suggest multiple modes of regulatory diversification, including alterations in transcription factor-binding specificity, incorporation of new gene targets into an existing regulatory system, and cooption of regulatory systems to control a different set of genes. We investigated one example in greater detail by measuring the in vitro activity of the S. cerevisiae transcription factor Rpn4p and its orthologs from Candida albicans and Neurospora crassa. Our results suggest that the DNA binding specificity of these proteins has coevolved with the sequences found upstream of the Rpn4p target genes and suggest that Rpn4p has a different function in N. crassa.

  4. Explosively launched spores of ascomycete fungi have drag-minimizing shapes

    PubMed Central

    Roper, Marcus; Pepper, Rachel E.; Brenner, Michael P.; Pringle, Anne

    2008-01-01

    The forcibly launched spores of ascomycete fungi must eject through several millimeters of nearly still air surrounding fruiting bodies to reach dispersive air flows. Because of their microscopic size, spores experience great fluid drag, and although this drag can aid transport by slowing sedimentation out of dispersive air flows, it also causes spores to decelerate rapidly after launch. We hypothesize that spores are shaped to maximize their range in the nearly still air surrounding fruiting bodies. To test this hypothesis we numerically calculate optimal spore shapesshapes of minimum drag for prescribed volumesand compare these shapes with real spore shapes taken from a phylogeny of >100 species. Our analysis shows that spores are constrained to remain within 1% of the minimum possible drag for their size. From the spore shapes we predict the speed of spore launch, and confirm this prediction through high-speed imaging of ejection in Neurospora tetrasperma. By reconstructing the evolutionary history of spore shapes within a single ascomycete family we measure the relative contributions of drag minimization and other shape determinants to spore shape evolution. Our study uses biomechanical optimization as an organizing principle for explaining shape in a mega-diverse group of species and provides a framework for future measurements of the forces of selection toward physical optima. PMID:19104035

  5. LIGNOCELLULOSE-DEGRADING ENZYMES PRODUCED BY THE ASCOMYCETE CONIOCHAETA LIGNIARIA AND RELATED SPECIES: APPLICATION FOR A LIGNOCELLULOSIC SUBSTRATE TREATMENT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lignocellulose-degrading microorganisms are of interest for biomass upgrading. In previous work, we isolated the ascomycete Coniochaeta ligniaria NRRL 30616 that metabolized phenolics and furans in lignocellulosic acid hydrolysates. This fungal isolate was investigated in the present work for the ...

  6. Freshwater ascomycetes: Alascospora evergladensis, a new genus and species from the Florida Everglades.

    PubMed

    Raja, Huzefa A; Violi, Helen A; Shearer, Carol A

    2010-01-01

    Alascospora evergladensis, a freshwater ascomycete collected from submerged dead petioles of Nymphaea odorata during a survey of aquatic fungi along a phosphorus gradient in the Florida Everglades, is described and illustrated as a new genus and species in the Pleosporales (Pleosporomycetidae, Dothideomycetes). The new fungus is unique among genera in the Pleosporales based on a combination of morphological characters that include light brown, translucent, membranous, ostiolate ascomata with dark, amorphous material irregularly deposited on the peridium, especially around the ostiole; globose, fissitunicate, thick-walled asci; septate pseudoparaphyses; and 1-septate ascospores that are hyaline when young, and surrounded by a hyaline gelatinous sheath that is wing-shaped in outline on each side of the ascospore. The sheath is distinctive in that it first expands in water and is translucent, then condenses and darkens around older ascospores, giving them a dark brown, verruculose appearance. PMID:20120226

  7. Bioconversion of (+)-valencene in submerged cultures of the ascomycete Chaetomium globosum.

    PubMed

    Kaspera, Rüdiger; Krings, Ulrich; Nanzad, Tsevegsuren; Berger, Ralf G

    2005-06-01

    Submerged cultures of the ascomycete Chaetomium globosum oxidised the exogenous sesquiterpene (+)-valencene to nootkatone via the stereoselective generation of alpha-nootkatol. Inhibition experiments suggested that the first introduction of oxygen, the rate-limiting step of the bioconversion, may have been catalysed by a cytochrome-P450-monooxygenase. However, nootkatone was not the final metabolite: further flavour-active and inactive, non-volatile oxidation products were identified. (+)-Valencene and the flavour-active mono-oxyfunctionalised transformation products, alpha-nootkatol, nootkatone, and valencene-11,12-epoxide accumulated preferably inside the fungal cells. Di- and poly-oxygenated products, such as nootkatone-11,12-epoxide, were found solely in the culture medium, indicating an active transport of these metabolites into the extracellular compartment during (+)-valencene detoxification. These metabolic properties may have contributed to the high tolerance of the fungus towards the exogenous hydrocarbon. PMID:15602686

  8. Genome mining of ascomycetous fungi reveals their genetic potential for ergot alkaloid production.

    PubMed

    Gerhards, Nina; Matuschek, Marco; Wallwey, Christiane; Li, Shu-Ming

    2015-06-01

    Ergot alkaloids are important as mycotoxins or as drugs. Naturally occurring ergot alkaloids as well as their semisynthetic derivatives have been used as pharmaceuticals in modern medicine for decades. We identified 196 putative ergot alkaloid biosynthetic genes belonging to at least 31 putative gene clusters in 31 fungal species by genome mining of the 360 available genome sequences of ascomycetous fungi with known proteins. Detailed analysis showed that these fungi belong to the families Aspergillaceae, Clavicipitaceae, Arthrodermataceae, Helotiaceae and Thermoascaceae. Within the identified families, only a small number of taxa are represented. Literature search revealed a large diversity of ergot alkaloid structures in different fungi of the phylum Ascomycota. However, ergot alkaloid accumulation was only observed in 15 of the sequenced species. Therefore, this study provides genetic basis for further study on ergot alkaloid production in the sequenced strains. PMID:25796201

  9. Analysis of Circadian Rhythms in the Basal Filamentous Ascomycete Pyronema confluens

    PubMed Central

    Traeger, Stefanie; Nowrousian, Minou

    2015-01-01

    Many organisms use circadian clocks to adapt to daily changes in the environment. Major insights into the molecular mechanisms of circadian oscillators have been gained through studies of the model organism Neurospora crassa; however, little is known about molecular components of circadian clocks in other fungi. An important part of the N. crassa circadian clock is the frequency (frq) gene, homologs of which can be found in Sordariomycetes, Dothideomycetes, and Leotiomycetes, but not Eurotiomycetes. Recently, we identified a frq homolog in Pyronema confluens, a member of the early-diverging Pezizomycete lineage of filamentous ascomycetes. The P. confluens FRQ shares many conserved domains with the N. crassa FRQ. However, there is no known morphological phenotype showing overt circadian rhythmicity in P. confluens. To investigate whether a molecular clock is present, we analyzed frq transcription in constant darkness, and found circadian oscillation of frq with a peak in the subjective morning. This rhythm was temperature compensated. To identify additional clock-controlled genes, we performed RNA sequencing of two time points (subjective morning and evening). Circadian expression of two morning-specific genes was verified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) over a full time course, whereas expression of two putative morning-specific and five putative evening-specific genes could not be verified as circadian. frq expression was synchronized, but not entrained by light. In summary, we have found evidence for two of the three main properties of circadian rhythms (free-running rhythm, temperature compensation) in P. confluens, suggesting that a circadian clock with rhythmically expressed frq is present in this basal filamentous ascomycete. PMID:26254031

  10. PCR primers that allow intergeneric differentiation of ascomycetes and their application to Verticillium spp.

    PubMed

    Li, K N; Rouse, D I; German, T L

    1994-12-01

    A pair of conserved PCR primers, designated NMS1 and NMS2, that amplify a region in the mitochondrial small rRNA gene region were designed for fungi belonging to the class Ascomycetes. These primers were tested with members of eight fungal genera (Aspergillus, Fusarium, Magnaporthe, Mycospharella, Neurospora, Saccharomyces, Sclerotinia, Verticillium) and 10 Verticillium species (Verticillium albo-atrum, Verticillium chlamydosporium, Verticillium cinnebarium, Verticillium dahliae, Verticillium fungicola, Verticillium lecanii, Verticillium lateritium, Verticillium nigrescens, Verticillium psaliotae, and Verticillium tricorpus). The primers were also tested with 35 isolates of V. dahliae obtained from diverse geographic areas and diverse hosts. The results of a restriction fragment length polymorphism analysis of the region amplified by the primers differentiated the genera examined and the results of a DNA sequence analysis of the amplified region differentiated the Verticillium species. Two Fusarium species were also differentiated by the results of the restriction fragment length polymorphism analysis. On the basis of the nucleotide sequences of the amplified regions, we obtained a pair of PCR primers that could be used to differentiate V. dahliae from the other fungal isolates tested, including V. albo-atrum, a closely related plant-pathogenic species. The V. dahliae-specific PCR primer may aid in more rapid and specific detection of the pathogen directly in plant and/or soil samples. PCR primers NMS1 and NMS2 may be used as potential mitochondrial markers for studying fungal cytoplasmic inheritance of ascomycetes and for identifying DNA probes that are informative at or below the genus level. PMID:7811072

  11. PCR primers that allow intergeneric differentiation of ascomycetes and their application to Verticillium spp.

    PubMed Central

    Li, K N; Rouse, D I; German, T L

    1994-01-01

    A pair of conserved PCR primers, designated NMS1 and NMS2, that amplify a region in the mitochondrial small rRNA gene region were designed for fungi belonging to the class Ascomycetes. These primers were tested with members of eight fungal genera (Aspergillus, Fusarium, Magnaporthe, Mycospharella, Neurospora, Saccharomyces, Sclerotinia, Verticillium) and 10 Verticillium species (Verticillium albo-atrum, Verticillium chlamydosporium, Verticillium cinnebarium, Verticillium dahliae, Verticillium fungicola, Verticillium lecanii, Verticillium lateritium, Verticillium nigrescens, Verticillium psaliotae, and Verticillium tricorpus). The primers were also tested with 35 isolates of V. dahliae obtained from diverse geographic areas and diverse hosts. The results of a restriction fragment length polymorphism analysis of the region amplified by the primers differentiated the genera examined and the results of a DNA sequence analysis of the amplified region differentiated the Verticillium species. Two Fusarium species were also differentiated by the results of the restriction fragment length polymorphism analysis. On the basis of the nucleotide sequences of the amplified regions, we obtained a pair of PCR primers that could be used to differentiate V. dahliae from the other fungal isolates tested, including V. albo-atrum, a closely related plant-pathogenic species. The V. dahliae-specific PCR primer may aid in more rapid and specific detection of the pathogen directly in plant and/or soil samples. PCR primers NMS1 and NMS2 may be used as potential mitochondrial markers for studying fungal cytoplasmic inheritance of ascomycetes and for identifying DNA probes that are informative at or below the genus level. Images PMID:7811072

  12. Eisosome Organization in the Filamentous AscomyceteAspergillus nidulans▿†

    PubMed Central

    Vangelatos, Ioannis; Roumelioti, Katerina; Gournas, Christos; Suarez, Teresa; Scazzocchio, Claudio; Sophianopoulou, Vicky

    2010-01-01

    Eisosomes are subcortical organelles implicated in endocytosis and have hitherto been described only in Saccharomyces cerevisiae. They comprise two homologue proteins, Pil1 and Lsp1, which colocalize with the transmembrane protein Sur7. These proteins are universally conserved in the ascomycetes. We identify in Aspergillus nidulans (and in all members of the subphylum Pezizomycotina) two homologues of Pil1/Lsp1, PilA and PilB, originating from a duplication independent from that extant in the subphylum Saccharomycotina. In the aspergilli there are several Sur7-like proteins in each species, including one strict Sur7 orthologue (SurG in A. nidulans). In A. nidulans conidiospores, but not in hyphae, the three proteins colocalize at the cell cortex and form tightly packed punctate structures that appear different from the clearly distinct eisosome patches observed in S. cerevisiae. These structures are assembled late during the maturation of conidia. In mycelia, punctate structures are present, but they are composed only of PilA, while PilB is diffused in the cytoplasm and SurG is located in vacuoles and endosomes. Deletion of each of the genes does not lead to any obvious growth phenotype, except for moderate resistance to itraconazole. We could not find any obvious association between mycelial (PilA) eisosome-like structures and endocytosis. PilA and SurG are necessary for conidial eisosome organization in ways that differ from those for their S. cerevisiae homologues. These data illustrate that conservation of eisosomal proteins within the ascomycetes is accompanied by a striking functional divergence. PMID:20693301

  13. Calnexin induces expansion of antigen-specific CD4+ T cells that confer immunity to fungal ascomycetes via conserved epitopes

    PubMed Central

    Wüthrich, Marcel; Brandhorst, Tristan T.; Sullivan, Thomas D.; Filutowicz, Hanna; Sterkel, Alana; Stewart, Douglas; Li, Mengyi; Lerksuthirat, Tassanee; LeBert, Vanessa; Shen, Zu Ting; Ostroff, Gary; Deepe, George S.; Hung, Chiung Yu; Cole, Garry; Walter, Jennifer A.; Jenkins, Marc K.; Klein, Bruce

    2015-01-01

    Fungal infections remain a threat due to the lack of broad spectrum fungal vaccines and protective antigens. Recent studies showed that attenuated Blastomyces dermatitidis confers protection via T cell recognition of an unknown, but conserved antigen. Using transgenic CD4+ T cells recognizing this antigen, we identify an amino acid determinant within the chaperone calnexin that is conserved across diverse fungal ascomycetes. Calnexin, typically an ER protein, also localizes to the surface of yeast, hyphae and spores. T cell epitope mapping unveiled a 13-residue sequence conserved across Ascomycota. Infection with divergent ascomycetes including dimorphic fungi, opportunistic molds, and the agent causing white nose syndrome in bats induces expansion of calnexin-specific CD4+ T cells. Vaccine delivery of calnexin in glucan particles induces fungal antigen-specific CD4+ T cell expansion and resistance to lethal challenge with multiple fungal pathogens. Thus, the immunogeneticity and conservation of calnexin make this fungal protein a promising vaccine target. PMID:25800545

  14. Calnexin induces expansion of antigen-specific CD4(+) T cells that confer immunity to fungal ascomycetes via conserved epitopes.

    PubMed

    Wüthrich, Marcel; Brandhorst, Tristan T; Sullivan, Thomas D; Filutowicz, Hanna; Sterkel, Alana; Stewart, Douglas; Li, Mengyi; Lerksuthirat, Tassanee; LeBert, Vanessa; Shen, Zu Ting; Ostroff, Gary; Deepe, George S; Hung, Chiung Yu; Cole, Garry; Walter, Jennifer A; Jenkins, Marc K; Klein, Bruce

    2015-04-01

    Fungal infections remain a threat due to the lack of broad-spectrum fungal vaccines and protective antigens. Recent studies showed that attenuated Blastomyces dermatitidis confers protection via T cell recognition of an unknown but conserved antigen. Using transgenic CD4(+) T cells recognizing this antigen, we identify an amino acid determinant within the chaperone calnexin that is conserved across diverse fungal ascomycetes. Calnexin, typically an ER protein, also localizes to the surface of yeast, hyphae, and spores. T cell epitope mapping unveiled a 13-residue sequence conserved across Ascomycota. Infection with divergent ascomycetes, including dimorphic fungi, opportunistic molds, and the agent causing white nose syndrome in bats, induces expansion of calnexin-specific CD4(+) T cells. Vaccine delivery of calnexin in glucan particles induces fungal antigen-specific CD4(+) T cell expansion and resistance to lethal challenge with multiple fungal pathogens. Thus, the immunogenicity and conservation of calnexin make this fungal protein a promising vaccine target. PMID:25800545

  15. Long-term experimental warming alters community composition of ascomycetes in Alaskan moist and dry arctic tundra.

    PubMed

    Semenova, Tatiana A; Morgado, Luis N; Welker, Jeffrey M; Walker, Marilyn D; Smets, Erik; Geml, József

    2015-01-01

    Arctic tundra regions have been responding to global warming with visible changes in plant community composition, including expansion of shrubs and declines in lichens and bryophytes. Even though it is well known that the majority of arctic plants are associated with their symbiotic fungi, how fungal community composition will be different with climate warming remains largely unknown. In this study, we addressed the effects of long-term (18 years) experimental warming on the community composition and taxonomic richness of soil ascomycetes in dry and moist tundra types. Using deep Ion Torrent sequencing, we quantified how OTU assemblage and richness of different orders of Ascomycota changed in response to summer warming. Experimental warming significantly altered ascomycete communities with stronger responses observed in the moist tundra compared with dry tundra. The proportion of several lichenized and moss-associated fungi decreased with warming, while the proportion of several plant and insect pathogens and saprotrophic species was higher in the warming treatment. The observed alterations in both taxonomic and ecological groups of ascomycetes are discussed in relation to previously reported warming-induced shifts in arctic plant communities, including decline in lichens and bryophytes and increase in coverage and biomass of shrubs. PMID:25522194

  16. Vesicular transport in Histoplasma capsulatum: an effective mechanism for trans-cell wall transfer of proteins and lipids in ascomycetes

    PubMed Central

    Albuquerque, Priscila Costa; Nakayasu, Ernesto S.; Rodrigues, Marcio L.; Frases, Susana; Casadevall, Arturo; Zancope-Oliveira, Rosely M.; Almeida, Igor C.; Nosanchuk, Joshua D.

    2008-01-01

    Vesicular secretion of macromolecules has recently been described in the basidiomycete Cryptococcus neoformans raising the question as to whether ascomycetes similarly utilize vesicles for transport. In the present study, we examine whether the clinically important ascomycete Histoplasma capsulatum produce vesicles and utilized these structures to secrete macromolecules. Transmission electron microscopy (TEM) show transcellular secretion of vesicles by yeast cells. Proteomic and lipidomic analyses of vesicles isolated from culture supernatants reveals a rich collection of macromolecules involved in diverse processes including metabolism, cell recycling, signaling, and virulence. The results demonstrate that H. capsulatum can utilize a trans-cell wall vesicular transport secretory mechanism to promote virulence. Additionally, TEM of supernatants collected from Candida albicans, Candida parapsilosis, Sporothrix schenckii, and Saccharomyces cerevisiae document that vesicles are similarly produced by additional ascomycetes. The vesicles from H. capsulatum react with immune serum from patients with histoplasmosis providing an association of the vesicular products with pathogenesis. The findings support the proposal that vesicular secretion is a general mechanism in fungi for the transport of macromolecules related to virulence and that this process could be a target for novel therapeutics. PMID:18419773

  17. "Hanseniaspora uvarum" the ultrastructural morphology of a rare ascomycete, isolated from oral thrush.

    PubMed

    Emmanouil-Nikoloussi, E; Kanellaki-Kyparissi, M; Papavassiliou, P; Koliakos, K; Dermentzopoulou, M; Foroglou, C

    1994-01-01

    Superficial fungal infections, including oral thrush, often affect aged full denture wearers and many individuals over 65 years old. The aim of this study was to examine the ultrastructural morphology of a very rare yeast, named Hanseniaspora uvarum/guillermondi, member of the Ascomycetes family, whose pathogenesis and behaviour is not widely known. The yeast was isolated from whitish lesions of the buccal mucosa of an 70 years old woman. The specimen was collected with a mouth swab and cultured in Sabourauds-Dextrose agar. The identification of the organism was performed on the Api 20C Aux system. The yeast colonies, after fixation in glutaraldehyde 3% for 1 hour were immersed in OsO4 1% solution for 1 hour and were "in tissue" stained with uranyl acetate. Ultrathin sections, were observed with TEM Jeol C x 100. Our ultrastructural observations showed that this yeast had a thick cell wall in which the outer surface appeared fuzzy. In some yeasts we observed multilayered intracytoplasmic membrane a figure which is not described as far as we know in any yeast. Many vacuoles were frequently observed in the cytoplasm and especially in the center of the oval shaped cells. Bilateral budding which form ascospores is identical for the morphology of this yeast. PMID:7994154

  18. A Putative Transcription Factor MYT2 Regulates Perithecium Size in the Ascomycete Gibberella zeae

    PubMed Central

    Lin, Yang; Son, Hokyoung; Min, Kyunghun; Lee, Jungkwan; Choi, Gyung Ja; Kim, Jin-Cheol; Lee, Yin-Won

    2012-01-01

    The homothallic ascomycete fungus Gibberella zeae is a plant pathogen that is found worldwide, causing Fusarium head blight (FHB) in cereal crops and ear rot of maize. Ascospores formed in fruiting bodies (i.e., perithecia) are hypothesized to be the primary inocula for FHB disease. Perithecium development is a complex cellular differentiation process controlled by many developmentally regulated genes. In this study, we selected a previously reported putative transcription factor containing the Myb DNA-binding domain MYT2 for an in-depth study on sexual development. The deletion of MYT2 resulted in a larger perithecium, while its overexpression resulted in a smaller perithecium when compared to the wild-type strain. These data suggest that MYT2 regulates perithecium size differentiation. MYT2 overexpression affected pleiotropic phenotypes including vegetative growth, conidia production, virulence, and mycotoxin production. Nuclear localization of the MYT2 protein supports its role as a transcriptional regulator. Transcriptional analyses of trichothecene synthetic genes suggest that MYT2 additionally functions as a suppressor for trichothecene production. This is the first study characterizing a transcription factor required for perithecium size differentiation in G. zeae, and it provides a novel angle for understanding sexual development in filamentous fungi. PMID:22649560

  19. Starmerella orientalis f.a., sp. nov., an ascomycetous yeast species isolated from flowers.

    PubMed

    Alimadadi, Nayyereh; Soudi, Mohammad Reza; Wang, Shi-An; Wang, Qi-Ming; Talebpour, Zahra; Bai, Feng-Yan

    2016-03-01

    Four strains of a novel ascomycetous yeast species were isolated from flowers in Iran and China. Phylogenetic analysis of the sequences of the ITS region (including 5.8S rRNA gene) and the LSU rRNA gene D1/D2 domains indicated that these strains belong to the Starmerella clade and show divergence from previously described species in this clade. Growth reactions on carbon and nitrogen sources were similar to those observed in related species of the Starmerella clade. Sexual reproduction was not observed after mating tests on different sporulation media. Based on physiological characteristics and phylogeny of rRNA gene sequences, the novel species is most closely related to Candida (iter. nom. Starmerella) powellii and Candida (iter. nom. Starmerella) floricola. It is therefore assigned to the genus Starmerella and described as Starmerella orientalis f.a., sp. nov. The type strain is SAM09T ( = IBRC-M 30204T = CBS 14142T). The MycoBank accession number is MB 814379. PMID:26780917

  20. Freshwater ascomycetes: two new species of Lindgomyces (Lindgomycetaceae, Pleosporales, Dothideomycetes) from Japan and USA.

    PubMed

    Raja, Huzefa A; Tanaka, Kazuaki; Hirayama, Kazuyuki; Miller, Andrew N; Shearer, Carol A

    2011-01-01

    During independent surveys of freshwater ascomycetes in Japan and USA two new species of Lindgomyces were collected from submerged wood in freshwater. These species are described and illustrated based on morphological data and phylogenetic relationships based on analyses of nuclear ribosomal sequence data (partial SSU and LSU, and ITS). Lindgomyces apiculatus, collected in Japan, is characterized by immersed to erumpent, globose to subglobose ascomata; fissitunicate, cylindrical to clavate asci; and fusiform, one-septate ascospores with acute ends and short terminal appendages. Lindgomyces lemonweirensis, collected in Wisconsin, USA, differs from L. apiculatus in having clavate to cymbiform asci and oblong to fusiform ascospores that are distinctively multiguttulate and surrounded by an oval, ephemeral gelatinous sheath. The new species formed a strongly supported clade within the family Lindgomycetaceae (Pleosporales, Dothideomycetes) based on analyses of combined SSU and LSU sequence data. In addition phylogenetic analyses with ITS sequence data support the establishment of the new taxa as separate species within Lindgomyces because they were separated from each other and other Lindgomyces species based on maximum likelihood bootstrap and Bayesian analyses. PMID:21700632

  1. Phylogenomic analysis of type I polyketide synthase genes in pathogenic and saprobic ascomycetes

    PubMed Central

    Kroken, Scott; Glass, N. Louise; Taylor, John W.; Yoder, O. C.; Turgeon, B. Gillian

    2003-01-01

    Fungal type I polyketides (PKs) are synthesized by PK synthases (PKSs) and include well known secondary metabolites such as the anticholesterol drug lovastatin and the potent natural carcinogen aflatoxin. Other type I PKs are known to be virulence factors for some plant pathogens and pigments such as melanin. In this study, a phylogenomic approach was used to investigate the origin and diversity of fungal genes encoding putative PKSs that are predicted to synthesize type I PKs. The resulting genealogy, constructed by using the highly conserved PKS ketosynthase (KS) domain, indicated that: (i) Species within subphylum Pezizomycotina (phylum Ascomycota) but not early diverging ascomycetes, like Saccharomyces cerevisiae (Saccharomycotina) or Schizosaccharomyces pombe (Taphrinomycotina), had large numbers (7–25) of PKS genes. (ii) Bacteria and fungi had separate groups of PKS genes; the few exceptions are the likely result of horizontal gene transfer from bacteria to various sublineages of fungi. (iii) The bulk of genes encoding fungal PKSs fell into eight groups. Four groups were predicted to synthesize variously reduced PKs, and four groups were predicted to make unreduced PKs. (iv) Species within different classes of Pezizomycotina shared the same groups of PKS genes. (v) Different fungal genomes shared few putative orthologous PKS genes, even between closely related genomes in the same class or genus. (vi) The discontinuous distributions of orthologous PKSs among fungal species can be explained by gene duplication, divergence, and gene loss; horizontal gene transfer among fungi does not need to be invoked. PMID:14676319

  2. Repeat-Associated Fission Yeast-Like Regional Centromeres in the Ascomycetous Budding Yeast Candida tropicalis

    PubMed Central

    Chatterjee, Gautam; Sankaranarayanan, Sundar Ram; Guin, Krishnendu; Thattikota, Yogitha; Padmanabhan, Sreedevi; Siddharthan, Rahul; Sanyal, Kaustuv

    2016-01-01

    The centromere, on which kinetochore proteins assemble, ensures precise chromosome segregation. Centromeres are largely specified by the histone H3 variant CENP-A (also known as Cse4 in yeasts). Structurally, centromere DNA sequences are highly diverse in nature. However, the evolutionary consequence of these structural diversities on de novo CENP-A chromatin formation remains elusive. Here, we report the identification of centromeres, as the binding sites of four evolutionarily conserved kinetochore proteins, in the human pathogenic budding yeast Candida tropicalis. Each of the seven centromeres comprises a 2 to 5 kb non-repetitive mid core flanked by 2 to 5 kb inverted repeats. The repeat-associated centromeres of C. tropicalis all share a high degree of sequence conservation with each other and are strikingly diverged from the unique and mostly non-repetitive centromeres of related Candida species—Candida albicans, Candida dubliniensis, and Candida lusitaniae. Using a plasmid-based assay, we further demonstrate that pericentric inverted repeats and the underlying DNA sequence provide a structural determinant in CENP-A recruitment in C. tropicalis, as opposed to epigenetically regulated CENP-A loading at centromeres in C. albicans. Thus, the centromere structure and its influence on de novo CENP-A recruitment has been significantly rewired in closely related Candida species. Strikingly, the centromere structural properties along with role of pericentric repeats in de novo CENP-A loading in C. tropicalis are more reminiscent to those of the distantly related fission yeast Schizosaccharomyces pombe. Taken together, we demonstrate, for the first time, fission yeast-like repeat-associated centromeres in an ascomycetous budding yeast. PMID:26845548

  3. Repeat-Associated Fission Yeast-Like Regional Centromeres in the Ascomycetous Budding Yeast Candida tropicalis.

    PubMed

    Chatterjee, Gautam; Sankaranarayanan, Sundar Ram; Guin, Krishnendu; Thattikota, Yogitha; Padmanabhan, Sreedevi; Siddharthan, Rahul; Sanyal, Kaustuv

    2016-02-01

    The centromere, on which kinetochore proteins assemble, ensures precise chromosome segregation. Centromeres are largely specified by the histone H3 variant CENP-A (also known as Cse4 in yeasts). Structurally, centromere DNA sequences are highly diverse in nature. However, the evolutionary consequence of these structural diversities on de novo CENP-A chromatin formation remains elusive. Here, we report the identification of centromeres, as the binding sites of four evolutionarily conserved kinetochore proteins, in the human pathogenic budding yeast Candida tropicalis. Each of the seven centromeres comprises a 2 to 5 kb non-repetitive mid core flanked by 2 to 5 kb inverted repeats. The repeat-associated centromeres of C. tropicalis all share a high degree of sequence conservation with each other and are strikingly diverged from the unique and mostly non-repetitive centromeres of related Candida species--Candida albicans, Candida dubliniensis, and Candida lusitaniae. Using a plasmid-based assay, we further demonstrate that pericentric inverted repeats and the underlying DNA sequence provide a structural determinant in CENP-A recruitment in C. tropicalis, as opposed to epigenetically regulated CENP-A loading at centromeres in C. albicans. Thus, the centromere structure and its influence on de novo CENP-A recruitment has been significantly rewired in closely related Candida species. Strikingly, the centromere structural properties along with role of pericentric repeats in de novo CENP-A loading in C. tropicalis are more reminiscent to those of the distantly related fission yeast Schizosaccharomyces pombe. Taken together, we demonstrate, for the first time, fission yeast-like repeat-associated centromeres in an ascomycetous budding yeast. PMID:26845548

  4. Mn(II) Oxidation by an Ascomycete Fungus is Linked to Superoxide Production During Asexual Reproduction

    SciTech Connect

    Hansel, Colleen M.; Zeiner, Carolyn A.; Santelli, Cara M.; Webb, Samuel M.

    2012-07-16

    Manganese (Mn) oxides are among the most reactive minerals within the environment, where they control the bioavailability of carbon, nutrients, and numerous metals. Although the ability of microorganisms to oxidize Mn(II) to Mn(III/IV) oxides is scattered throughout the bacterial and fungal domains of life, the mechanism and physiological basis for Mn(II) oxidation remains an enigma. Here, we use a combination of compound-specific chemical assays, microspectroscopy, and electron microscopy to show that a common Ascomycete filamentous fungus, Stilbella aciculosa, oxidizes Mn(II) to Mn oxides by producing extracellular superoxide during cell differentiation. The reactive Mn oxide phase birnessite and the reactive oxygen species superoxide and hydrogen peroxide are colocalized at the base of asexual reproductive structures. Mn oxide formation is not observed in the presence of superoxide scavengers (e.g., Cu) and inhibitors of NADPH oxidases (e.g., diphenylene iodonium chloride), enzymes responsible for superoxide production and cell differentiation in fungi. Considering the recent identification of Mn(II) oxidation by NADH oxidase-based superoxide production by a common marine bacterium (Roseobacter sp.), these results introduce a surprising homology between some prokaryotic and eukaryotic organisms in the mechanisms responsible for Mn(II) oxidation, where oxidation appears to be a side reaction of extracellular superoxide production. Given the versatility of superoxide as a redox reactant and the widespread ability of fungi to produce superoxide, this microbial extracellular superoxide production may play a central role in the cycling and bioavailability of metals (e.g., Hg, Fe, Mn) and carbon in natural systems.

  5. Functional properties and differential mode of regulation of the nitrate transporter from a plant symbiotic ascomycete

    PubMed Central

    Montanini, Barbara; Viscomi, Arturo R.; Bolchi, Angelo; Martin, Yusé; Siverio, José M.; Balestrini, Raffaella; Bonfante, Paola; Ottonello, Simone

    2005-01-01

    Nitrogen assimilation by plant symbiotic fungi plays a central role in the mutualistic interaction established by these organisms, as well as in nitrogen flux in a variety of soils. In the present study, we report on the functional properties, structural organization and distinctive mode of regulation of TbNrt2 (Tuber borchii NRT2 family transporter), the nitrate transporter of the mycorrhizal ascomycete T. borchii. As revealed by experiments conducted in a nitrate-uptake-defective mutant of the yeast Hansenula polymorpha, TbNrt2 is a high-affinity transporter (Km=4.7 μM nitrate) that is bispecific for nitrate and nitrite. It is expressed in free-living mycelia and in mycorrhizae, where it preferentially accumulates in the plasma membrane of root-contacting hyphae. The TbNrt2 mRNA, which is transcribed from a single-copy gene clustered with the nitrate reductase gene in the T. borchii genome, was specifically up-regulated following transfer of mycelia to nitrate- (or nitrite)-containing medium. However, at variance with the strict nitrate-dependent induction commonly observed in other organisms, TbNrt2 was also up-regulated (at both the mRNA and the protein level) following transfer to a nitrogen-free medium. This unusual mode of regulation differs from that of the adjacent nitrate reductase gene, which was expressed at basal levels under nitrogen deprivation conditions and required nitrate for induction. The functional and expression properties, described in the present study, delineate TbNrt2 as a versatile transporter that may be especially suited to cope with the fluctuating (and often low) mineral nitrogen concentrations found in most natural, especially forest, soils. PMID:16201972

  6. Asexual Cephalosporin C Producer Acremonium chrysogenum Carries a Functional Mating Type Locus▿

    PubMed Central

    Pöggeler, Stefanie; Hoff, Birgit; Kück, Ulrich

    2008-01-01

    Acremonium chrysogenum, the fungal producer of the pharmaceutically relevant β-lactam antibiotic cephalosporin C, is classified as asexual because no direct observation of mating or meiosis has yet been reported. To assess the potential of A. chrysogenum for sexual reproduction, we screened an expressed sequence tag library from A. chrysogenum for the expression of mating type (MAT) genes, which are the key regulators of sexual reproduction. We identified two putative mating type genes that are homologues of the α-box domain gene, MAT1-1-1 and MAT1-1-2, encoding an HPG domain protein defined by the presence of the three invariant amino acids histidine, proline, and glycine. In addition, cDNAs encoding a putative pheromone receptor and pheromone-processing enzymes, as well as components of a pheromone response pathway, were found. Moreover, the entire A. chrysogenum MAT1-1 (AcMAT1-1) gene and regions flanking the MAT region were obtained from a genomic cosmid library, and sequence analysis revealed that in addition to AcMAT1-1-1 and AcMAT1-1-2, the AcMAT1-1 locus comprises a third mating type gene, AcMAT1-1-3, encoding a high-mobility-group domain protein. The α-box domain sequence of AcMAT1-1-1 was used to determine the phylogenetic relationships of A. chrysogenum to other ascomycetes. To determine the functionality of the AcMAT1-1 locus, the entire MAT locus was transferred into a MAT deletion strain of the heterothallic ascomycete Podospora anserina (the PaΔMAT strain). After fertilization with a P. anserina MAT1-2 (MAT+) strain, the corresponding transformants developed fruiting bodies with mature ascospores. Thus, the results of our functional analysis of the AcMAT1-1 locus provide strong evidence to hypothesize a sexual cycle in A. chrysogenum. PMID:18689517

  7. Freshwater ascomycetes: Minutisphaera (Dothideomycetes) revisited, including one new species from Japan.

    PubMed

    Raja, Huzefa A; Oberlies, Nicholas H; Figueroa, Mario; Tanaka, Kazuaki; Hirayama, Kazuyuki; Hashimoto, Akira; Miller, Andrew N; Zelski, Steven E; Shearer, Carol A

    2013-01-01

    During investigations of freshwater ascomycetes we found one interesting taxon from Aomori (Japan), as well as three additional taxa from North Carolina (USA), which were morphologically similar to Minutisphaera, a recently described freshwater fungus in the Dothideomycetes. The ascomata of all the collections bore dark hair-like structures around the ostiolar region, obovoid to obclavate bitunicate asci, and one to three septate hyaline to brown ascospores with a sheath (in material from Japan), and with both sheath and appendages (in material from the USA). The apothecial ascomata of these taxa, however, differ from those of the type species of the genus, which are perithecial. Two collections of Minutisphaera-like fungi from the USA were morphologically quite similar but differed in ascospore size. To assess the phylogenetic affinities of Minutisphaera-like taxa with the type species, M. fimbriatispora, we sequenced 18S and 28S nrDNA of five newly collected strains of Minutisphaera. We also sequenced the nrDNA for the entire internal transcribed spacer region of 10 strains to assess interspecific and intraspecific variation with M. fimbriatispora. Additionally we examined the secondary metabolite profiles of two strains from USA. Based on maximum likelihood and Bayesian analyses of combined 18S and 28S, and separate ITS sequences, as well as examination of morphology, we describe and illustrate a new species, M. japonica. One collection from North Carolina is confirmed as M. fimbriatispora, while two other collections are Minutisphaera-like fungi that had a number of similar diagnostic morphological characters but differed only slightly in ascospore sizes. The phylogeny inferred from the internal transcribed spacer region suggested that two out of the three North Carolina collections may be novel and perhaps cryptic species within Minutisphaera. Organic extracts of Minutisphaera from USA, M. fimbriatispora (G155-1) and Minutisphaera-like taxon (G156-1), revealed the presence of palmitic acid and (E)-hexadec-9-en-1-ol as major chemical constituents. We discuss the placement of the Minutisphaera clade within the Dothideomycetes. The description of the genus Minutisphaera is emended to accommodate M. japonica within Minutisphaera. PMID:23709484

  8. RAPID IDENTFICATION OF ASCOMYCETOUS YEASTS FROM CLINICAL SPECIMENS BY A MOLECULAR-BASED FLOW CYTOMETRY METHOD AND COMPARISION WITH IDENTIFICATIONS FROM PHENOTYPIC ASSAYS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was designed to compare the identification of ascomycetous yeasts recovered from clinical specimens by using phenotypic assays (PA) and a molecular flow cytometric (FC) method. LSU rRNA D1/D2 gene sequence analysis was also performed and served as the reference for correct strain identif...

  9. Simulated aerial sprays for field cage evaluation of Beauveria bassiana and Metarhizium brunneum (Ascomycetes: Hypocreales) against Anabrus simplex (Orthoptera: Tettigoniidae) in Montana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field efficacy of the entomopathogenic Ascomycete Beauveria bassiana strain GHA and Metarhizium brunneum strain F52 was evaluated against nymphs of the Mormon cricket, Anabrus simplex. Fungi were applied with a new apparatus that allows simulated aerial sprays to 0.1m2 areas in the field. The Mormon...

  10. Identification of heavy metal regulated genes from the root associated ascomycete Cadophora finlandica using a genomic microarray.

    PubMed

    Gorfer, Markus; Persak, Helene; Berger, Harald; Brynda, Sabine; Bandian, Dragana; Strauss, Joseph

    2009-12-01

    The ascomycete Cadophora finlandica, which can form mycorrhizas with ectomycorrhizal and ericoid hosts, is commonly found in heavy metal polluted soils. To understand the selective advantage of this organism at contaminated sites heavy metal regulated genes from C. finlandica were investigated. For gene identification a strategy based on a genomic microarray was chosen, which allows a rapid, genome-wide screening in genetically poorly characterized organisms. In a preliminary screen eleven plasmids covering eight distinct genomic regions and encoding a total of ten Cd-regulated genes were identified. Northern analyses with RNA from C. finlandica grown in the presence of either Cd, Pb or Zn revealed different transcription patterns in response to the heavy metals present in the growth medium. The Cd-regulated genes are predicted to encode several extracellular proteins with unknown functions, transporters, a centaurin-type regulator of intracellular membrane trafficking, a GNAT-family acetyltransferase and a B-type cyclin. PMID:19770041

  11. Exploring laccase-like multicopper oxidase genes from the ascomycete Trichoderma reesei: a functional, phylogenetic and evolutionary study

    PubMed Central

    2010-01-01

    Background The diversity and function of ligninolytic genes in soil-inhabiting ascomycetes has not yet been elucidated, despite their possible role in plant litter decay processes. Among ascomycetes, Trichoderma reesei is a model organism of cellulose and hemicellulose degradation, used for its unique secretion ability especially for cellulase production. T. reesei has only been reported as a cellulolytic and hemicellulolytic organism although genome annotation revealed 6 laccase-like multicopper oxidase (LMCO) genes. The purpose of this work was i) to validate the function of a candidate LMCO gene from T. reesei, and ii) to reconstruct LMCO phylogeny and perform evolutionary analysis testing for positive selection. Results After homologous overproduction of a candidate LMCO gene, extracellular laccase activity was detected when ABTS or SRG were used as substrates, and the recombinant protein was purified to homogeneity followed by biochemical characterization. The recombinant protein, called TrLAC1, has a molecular mass of 104 kDa. Optimal temperature and pH were respectively 40-45°C and 4, by using ABTS as substrate. TrLAC1 showed broad pH stability range of 3 to 7. Temperature stability revealed that TrLAC1 is not a thermostable enzyme, which was also confirmed by unfolding studies monitored by circular dichroism. Evolutionary studies were performed to shed light on the LMCO family, and the phylogenetic tree was reconstructed using maximum-likelihood method. LMCO and classical laccases were clearly divided into two distinct groups. Finally, Darwinian selection was tested, and the results showed that positive selection drove the evolution of sequences leading to well-known laccases involved in ligninolysis. Positively-selected sites were observed that could be used as targets for mutagenesis and functional studies between classical laccases and LMCO from T. reesei. Conclusions Homologous production and evolutionary studies of the first LMCO from the biomass-degrading fungus T. reesei gives new insights into the physicochemical parameters and biodiversity in this family. PMID:20735824

  12. The nucleotide sequences of the 5 S rRNAs of seven molds and a yeast and their use in studying ascomycete phylogeny.

    PubMed Central

    Chen, M W; Anné, J; Volckaert, G; Huysmans, E; Vandenberghe, A; De Wachter, R

    1984-01-01

    The sequences of the 5 S rRNAs isolated from 8 ascomycete species belonging to the genera Aspergillus, Penicillium, Acremonium and Candida are reported. Two of the examined strains each yielded a mixture of 3 slightly different 5 S RNAs, which were individually sequenced after fractionation. A previously published sequence for Aspergillus nidulans 5 S RNA was found to contain errors. Reconstruction of an evolutionary tree based on 5 S RNA sequences showed that the 16 presently examined ascomycetes form three clusters. The same threefold partition can be observed in the secondary structure pattern, each cluster showing a slightly different variant of the general 5-helix model for 5 S rRNA (De Wachter, Chen and Vandenberghe (1982) Biochimie 64, 311-329), and different sets of secondary structure equilibrium forms in helices C and E of the aforementioned model. Images PMID:6429642

  13. A new pullulan and a branched (1-->3)-, (1-->6)-linked beta-glucan from the lichenised ascomycete Teloschistes flavicans.

    PubMed

    Reis, Rodrigo A; Tischer, Cesar A; Gorin, Philip A J; Iacomini, Marcello

    2002-04-23

    The polysaccharides formed on hot alkaline extraction of the ascomycetous lichen Teloschistes flavicans were fractionated to give two glucans, which were characterised by methylation analysis and 1D and 2D NMR spectroscopy. One was a branched beta-glucan containing (1-->3) and (1-->6) linkages, a structure which is more typical of basidiomycetes rather than ascomycetes, which have linear glucans. The other was an alpha-glucan with alternating (1-->4) and (1-->6) linkages, found for the first time in Nature. This structure can be classified as a pullulan, which has been isolated from the fungi Aureobasidium pullulans, Tremella mesenterica, and Cyttaria harioti, but has different ratios of the component glycosidic linkages. The significance of the presence of the isolated alpha- and beta-glucans is discussed. PMID:12023070

  14. Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties

    SciTech Connect

    Osipov, Evgeny; Kittl, Roman; Shleev, Sergey; Dorovatovsky, Pavel; Tikhonova, Tamara; Popov, Vladimir

    2014-11-01

    The structures of the ascomycetous B. aclada laccase and its L499M T1-site mutant have been solved at 1.7 Å resolution. The mutant enzyme shows a 140 mV lower redox potential of the type 1 copper and altered kinetic behaviour. The wild type and the mutant have very similar structures, which makes it possible to relate the changes in the redox potential to the L499M mutation Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. These enzymes contain four Cu atoms per molecule organized into three sites: T1, T2 and T3. In all laccases, the T1 copper ion is coordinated by two histidines and one cysteine in the equatorial plane and is covered by the side chains of hydrophobic residues in the axial positions. The redox potential of the T1 copper ion influences the enzymatic reaction and is determined by the nature of the axial ligands and the structure of the second coordination sphere. In this work, the laccase from the ascomycete Botrytis aclada was studied, which contains conserved Ile491 and nonconserved Leu499 residues in the axial positions. The three-dimensional structures of the wild-type enzyme and the L499M mutant were determined by X-ray crystallography at 1.7 Å resolution. Crystals suitable for X-ray analysis could only be grown after deglycosylation. Both structures did not contain the T2 copper ion. The catalytic properties of the enzyme were characterized and the redox potentials of both enzyme forms were determined: E{sub 0} = 720 and 580 mV for the wild-type enzyme and the mutant, respectively. Since the structures of the wild-type and mutant forms are very similar, the change in the redox potential can be related to the L499M mutation in the T1 site of the enzyme.

  15. Removal of chlorophenolic derivatives by soil isolated ascomycete of Paraconiothyrium variabile and studying the role of its extracellular laccase.

    PubMed

    Forootanfar, Hamid; Movahednia, Mohammad Mehdi; Yaghmaei, Soheila; Tabatabaei-Sameni, Minoosadat; Rastegar, Hossein; Sadighi, Armin; Faramarzi, Mohammad Ali

    2012-03-30

    The ability of Paraconiothyrium variabile, a laccase producing ascomycete recently isolated from soil, was studied to eliminate chlorophenol derivatives in submerged culture medium. Among the tested compounds, ρ-chlorophenol (ρ-CP) and pentachlorophenol (PCP) were found to have minimum and maximum toxic effects, respectively, on the growth of the microorganism and at the same time high and low bioelimination percentages. The fungal strain was able to remove 86% of ρ-CP (with initial concentration of 40 mg l(-1)) and 56% of 2,4-dichlorophenol (2,4-DCP; with same concentration as ρ-CP) after 9 days of incubation while no elimination was observed in the presence of 2,4,6-trichlorophenol (2,4,6-TCP) and PCP. Monitoring of laccase production level in the fermentation broth together with pollutant removal confirmed the key role of this copper-containing oxidase in chlorophenol derivatives elimination. The type of laccase inducer (guaiacol) and its final concentration (250 μM) and also initial pH of the fermentation broth (pH=5.5) in the elimination of ρ-CP increased the final removal yield from 86% to 94.3%. PMID:22277342

  16. Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi.

    PubMed

    Gacura, Matthew D; Sprockett, Daniel D; Heidenreich, Bess; Blackwood, Christopher B

    2016-04-01

    Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. PMID:26899925

  17. Genomic Analysis of an Ascomycete Fungus from the Rice Planthopper Reveals How It Adapts to an Endosymbiotic Lifestyle

    PubMed Central

    Fan, Hai-Wei; Noda, Hiroaki; Xie, Hong-Qing; Suetsugu, Yoshitaka; Zhu, Qian-Hua; Zhang, Chuan-Xi

    2015-01-01

    A number of sap-sucking insects harbor endosymbionts, which are thought to play an important role in the development of their hosts. One of the most important rice pests, the brown planthopper (BPH), Nilaparvata lugens (Stål), harbors an obligatory yeast-like symbiont (YLS) that cannot be cultured in vitro. Genomic information on this YLS would be useful to better understand its evolution. In this study, we performed genome sequencing of the YLS using both 454 and Illumina approaches, generating a draft genome that shows a slightly smaller genome size and relatively higher GC content than most ascomycete fungi. A phylogenomic analysis of the YLS supported its close relationship with insect pathogens. We analyzed YLS-specific genes and the categories of genes that are likely to have changed in the YLS during its evolution. The loss of mating type locus demonstrated in the YLS sheds light on the evolution of eukaryotic symbionts. This information about the YLS genome provides a helpful guide for further understanding endosymbiotic associations in hemiptera and the symbiotic replacement of ancient bacteria with a multifunctional YLS seems to have been a successful change. PMID:26338189

  18. Primer Sets Developed To Amplify Conserved Genes from Filamentous Ascomycetes Are Useful in Differentiating Fusarium Species Associated with Conifers

    PubMed Central

    Donaldson, G. C.; Ball, L. A.; Axelrood, P. E.; Glass, N. L.

    1995-01-01

    We examined the usefulness of primer sets designed to amplify introns within conserved genes in filamentous ascomycetes to differentiate 35 isolates representing six different species of Fusarium commonly found in association with conifer seedlings. We analyzed restriction fragment length polymorphisms (RFLP) in five amplified PCR products from each Fusarium isolate. The primers used in this study were constructed on the basis of sequence information from the H3, H4, and (beta)-tubulin genes in Neurospora crassa. Primers previously developed for the intergenic transcribed spacer region of the ribosomal DNA were also used. The degree of interspecific polymorphism observed in the PCR products from the six Fusarium species allowed differentiation by a limited number of amplifications and restriction endonuclease digestions. The level of intraspecific RFLP variation in the five PCR products was low in both Fusarium proliferatum and F. avenaceum but was high in a population sample of F. oxysporum isolates. Clustering of the 35 isolates by statistical analyses gave similar dendrograms for H3, H4, and (beta)-tubulin RFLP analysis, but a dendrogram produced by intergenic transcribed spacer analysis varied in the placement of some F. oxysporum isolates. PMID:16534991

  19. Genomic Analysis of an Ascomycete Fungus from the Rice Planthopper Reveals How It Adapts to an Endosymbiotic Lifestyle.

    PubMed

    Fan, Hai-Wei; Noda, Hiroaki; Xie, Hong-Qing; Suetsugu, Yoshitaka; Zhu, Qian-Hua; Zhang, Chuan-Xi

    2015-09-01

    A number of sap-sucking insects harbor endosymbionts, which are thought to play an important role in the development of their hosts. One of the most important rice pests, the brown planthopper (BPH), Nilaparvata lugens (Stl), harbors an obligatory yeast-like symbiont (YLS) that cannot be cultured in vitro. Genomic information on this YLS would be useful to better understand its evolution. In this study, we performed genome sequencing of the YLS using both 454 and Illumina approaches, generating a draft genome that shows a slightly smaller genome size and relatively higher GC content than most ascomycete fungi. A phylogenomic analysis of the YLS supported its close relationship with insect pathogens. We analyzed YLS-specific genes and the categories of genes that are likely to have changed in the YLS during its evolution. The loss of mating type locus demonstrated in the YLS sheds light on the evolution of eukaryotic symbionts. This information about the YLS genome provides a helpful guide for further understanding endosymbiotic associations in hemiptera and the symbiotic replacement of ancient bacteria with a multifunctional YLS seems to have been a successful change. PMID:26338189

  20. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea

    PubMed Central

    Ding, Zhong-Tao; Zhang, Zhi; Luo, Di; Zhou, Jin-Yan; Zhong, Juan; Yang, Jie; Xiao, Liang; Shu, Dan; Tan, Hong

    2015-01-01

    The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+)-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers) and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain. PMID:25955649

  1. Functional characterization of sucrose non-fermenting 1 protein kinase complex genes in the Ascomycete Fusarium graminearum.

    PubMed

    Yu, Jungheon; Son, Hokyoung; Park, Ae Ran; Lee, Seung-Ho; Choi, Gyung Ja; Kim, Jin-Cheol; Lee, Yin-Won

    2014-02-01

    Sucrose non-fermenting 1 (SNF1) protein kinase complex is a heterotrimer that functions in energy homeostasis in eukaryotes by regulating transcription of glucose-repressible genes. Our previous study revealed that SNF1 of the homothallic ascomycete fungus Fusarium graminearum plays important roles in vegetative growth, sexual development, and virulence. In this study, we further identified the components of the SNF1 complex in F. graminearum and characterized their functions. We found that the SNF1 complex in F. graminearum consists of one alpha subunit (FgSNF1), one beta subunit (FgGAL83), and one gamma subunit (FgSNF4). Deletion of Fggal83 and Fgsnf4 resulted in alleviated phenotype changes in vegetative growth and sexual development as compared to those of the Fgsnf1 deletion mutant. However, all of the single, double, and triple deletion mutants among Fgsnf1, Fggal83, and Fgsnf4 had similar levels of decreased virulence. In addition, there was no synergistic effect of the mutant (single, double, or triple deletions of SNF1 complex component genes) phenotypes except for sucrose utilization. In this study, we revealed that FgSNF1 is mainly required for SNF1 complex functions, and the other two SNF1 complex components have adjunctive roles with FgSNF1 in sexual development and vegetative growth but have a major role in virulence in F. graminearum. PMID:24057127

  2. Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi

    DOE PAGESBeta

    Gacura, Matthew D.; Sprockett, Daniel D.; Heidenreich, Bess; Blackwood, Christopher B.

    2016-02-17

    Here, fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). In this studywe developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from40 fungal genomes. The primerswere used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senescedmore » tree leaves from different forest ecosystems, followed by cloning and sequencing. Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communitieswere up to 39% Basidiomycetes. Analysis of leaf samples indicated that both forest stand and ecosystemtype were important in structuring fungal communities. However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes.« less

  3. Nutritional physiology of a rock-inhabiting, model microcolonial fungus from an ancestral lineage of the Chaetothyriales (Ascomycetes).

    PubMed

    Nai, Corrado; Wong, Helen Y; Pannenbecker, Annette; Broughton, William J; Benoit, Isabelle; de Vries, Ronald P; Gueidan, Cécile; Gorbushina, Anna A

    2013-07-01

    Rock-inhabiting black fungi [also microcolonial or meristematic fungi (MCF)] are a phylogenetically diverse group of melanised ascomycetes with distinctive morphological features that confer extensive stress tolerance and permit survival in hostile environments. The MCF strain A95 Knufia petricola (syn. Sarcinomyces petricola) belongs to an ancestral lineage of the order Chaetothyriales (class Eurotiomycetes). K. petricola strain A95 is a rock-inhabiting MCF and its growth requirements were studied using the 96-well plate-based Biolog System under ∼1070 different conditions (osmotic stress, pH growth optima, growth factor requirements and nutrient catabolism). A95 is an osmotolerant, oligotrophic MCF that grows best around pH 5. Remarkably, A95 shows metabolic activity in the absence of added nitrogen, phosphorus or sulphur. Correlations could be drawn between the known nutrient requirements of A95 and what probably is available in sub-aerial systems (rock and other material surfaces). Detailed knowledge of A95's metabolic requirements allowed formulation of a synthetic medium that supports strong fungal growth. PMID:23587800

  4. High-Coverage ITS Primers for the DNA-Based Identification of Ascomycetes and Basidiomycetes in Environmental Samples

    PubMed Central

    Yamamoto, Satoshi; Sato, Hirotoshi

    2012-01-01

    The kingdom Fungi is estimated to include 1.5 million or more species, playing key roles as decomposers, mutualists, and parasites in every biome on the earth. To comprehensively understand the diversity and ecology of this huge kingdom, DNA barcoding targeting the internal transcribed spacer (ITS) region of the nuclear ribosomal repeat has been regarded as a prerequisite procedure. By extensively surveying ITS sequences in public databases, we designed new ITS primers with improved coverage across diverse taxonomic groups of fungi compared to existing primers. An in silico analysis based on public sequence databases indicated that the newly designed primers matched 99% of ascomycete and basidiomycete ITS taxa (species, subspecies or varieties), causing little taxonomic bias toward either fungal group. Two of the newly designed primers could inhibit the amplification of plant sequences and would enable the selective investigation of fungal communities in mycorrhizal associations, soil, and other types of environmental samples. Optimal PCR conditions for the primers were explored in an in vitro investigation. The new primers developed in this study will provide a basis for ecological studies on the diversity and community structures of fungi in the era of massive DNA sequencing. PMID:22808280

  5. Xylogone ganodermophthora sp. nov., an ascomycetous pathogen causing yellow rot on cultivated mushroom Ganoderma lucidum in Korea.

    PubMed

    Kang, Hyo-Jung; Sigler, Lynne; Lee, Jungkwan; Gibas, Connie Fe C; Yun, Sung-Hwan; Lee, Yin-Won

    2010-01-01

    Yellow rot, caused by an ascomycetous fungus having a distinctive arthroconidial anamorph, is the most destructive disease of cultivated Ganoderma lucidum in Korea, but the identity of the yellow rot pathogen (YRP) remains uncertain. Isolates have been identified as Xylogone sphaerospora (with putative anamorph Sporendonema purpurascens) or as Arthrographis cuboidea. Therefore we used morphological features, pathogenicity tests and phylogenetic analyses of DNA sequences from the nuclear ribosomal genes, including partial small subunit and internal transcribed spacer regions, and from the gene encoding RNA polymerase second largest subunit to evaluate the relationship between YRP isolates and these species. YRP isolates formed a distinct subgroup within a clade that included X. sphaerospora, A. cuboidea and Scytalidium lignicola, the type species of Scytalidium, but the disposition of the clade within the Leotiomycetes was uncertain. We describe Xylogone ganodermophthora sp. nov. and Scytalidium ganodermophthorum sp. nov. for the teleomorph and anamorph of YRP respectively. Arthrographis cuboidea is reclassified as Scytalidium cuboideum comb. nov., and the anamorph of X. sphaerospora is named Scytalidium sphaerosporum sp. nov. In pathogenicity tests only X. ganodermophthora caused disease in Ganoderma lucidum. Amplified fragment length polymorphism analyses showed that X. ganodermophthora populations from diseased fruiting bodies or from oak wood in Korea consisted of two clonal groups. PMID:20943517

  6. NMR analysis of the binding mode of two fungal endo-β-1,4-mannanases from GH5 and GH26 families.

    PubMed

    Marchetti, Roberta; Berrin, Jean-Guy; Couturier, Marie; Ul Qader, Shah Ali; Molinaro, Antonio; Silipo, Alba

    2015-12-15

    The enzymatic digestion of the main components of lignocellulosic biomass, including plant cell wall mannans, constitutes a fundamental step in the renewable biofuel production, with great potential benefit in the industrial field. Despite several reports of X-ray structures of glycoside hydrolases, how polysaccharides are specifically recognized and accommodated in the enzymes binding site still remains a pivotal matter of research. Within this frame, NMR spectroscopic techniques provide key binding information, complementing and/or enhancing the structural view by X-ray crystallography. Here we provide deep insights into the binding mode of two endo-β-1,4 mannanases from the coprophilous ascomycete Podospora anserina, PaMan26A and PaMan5A, involved in the hydrolysis of plant cell wall mannans and heteromannans. The investigation at a molecular level of the interaction between the wild-type enzymes and inactive mutants with manno-oligosaccharides, revealed a different mode of action among the two glycoside hydrolases most likely due to the presence of the additional and peculiar -4 subsite in the PaMan26A binding pocket. PMID:26567779

  7. Genes That Bias Mendelian Segregation

    PubMed Central

    Grognet, Pierre; Lalucque, Hervé; Malagnac, Fabienne; Silar, Philippe

    2014-01-01

    Mendel laws of inheritance can be cheated by Meiotic Drive Elements (MDs), complex nuclear genetic loci found in various eukaryotic genomes and distorting segregation in their favor. Here, we identify and characterize in the model fungus Podospora anserina Spok1 and Spok2, two MDs known as Spore Killers. We show that they are related genes with both spore-killing distorter and spore-protecting responder activities carried out by the same allele. These alleles act as autonomous elements, exert their effects independently of their location in the genome and can act as MDs in other fungi. Additionally, Spok1 acts as a resistance factor to Spok2 killing. Genetical data and cytological analysis of Spok1 and Spok2 localization during the killing process suggest a complex mode of action for Spok proteins. Spok1 and Spok2 belong to a multigene family prevalent in the genomes of many ascomycetes. As they have no obvious cellular role, Spok1 and Spok2 Spore Killer genes represent a novel kind of selfish genetic elements prevalent in fungal genome that proliferate through meiotic distortion. PMID:24830502

  8. Contrasting Diversity and Host Association of Ectomycorrhizal Basidiomycetes versus Root-Associated Ascomycetes in a Dipterocarp Rainforest

    PubMed Central

    Sato, Hirotoshi; Tanabe, Akifumi S.; Toju, Hirokazu

    2015-01-01

    Root-associated fungi, including ectomycorrhizal and root-endophytic fungi, are among the most diverse and important belowground plant symbionts in dipterocarp rainforests. Our study aimed to reveal the biodiversity, host association, and community structure of ectomycorrhizal Basidiomycota and root-associated Ascomycota (including root-endophytic Ascomycota) in a lowland dipterocarp rainforest in Southeast Asia. The host plant chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) region and fungal internal transcribed spacer 2 (ITS2) region were sequenced using tag-encoded, massively parallel 454 pyrosequencing to identify host plant and root-associated fungal taxa in root samples. In total, 1245 ascomycetous and 127 putative ectomycorrhizal basidiomycetous taxa were detected from 442 root samples. The putative ectomycorrhizal Basidiomycota were likely to be associated with closely related dipterocarp taxa to greater or lesser extents, whereas host association patterns of the root-associated Ascomycota were much less distinct. The community structure of the putative ectomycorrhizal Basidiomycota was possibly more influenced by host genetic distances than was that of the root-associated Ascomycota. This study also indicated that in dipterocarp rainforests, root-associated Ascomycota were characterized by high biodiversity and indistinct host association patterns, whereas ectomycorrhizal Basidiomycota showed less biodiversity and a strong host phylogenetic preference for dipterocarp trees. Our findings lead to the working hypothesis that root-associated Ascomycota, which might be mainly represented by root-endophytic fungi, have biodiversity hotspots in the tropics, whereas biodiversity of ectomycorrhizal Basidiomycota increases with host genetic diversity. PMID:25884708

  9. The mating-type chromosome in the filamentous ascomycete Neurospora tetrasperma represents a model for early evolution of sex chromosomes.

    PubMed

    Menkis, Audrius; Jacobson, David J; Gustafsson, Tim; Johannesson, Hanna

    2008-03-01

    We combined gene divergence data, classical genetics, and phylogenetics to study the evolution of the mating-type chromosome in the filamentous ascomycete Neurospora tetrasperma. In this species, a large non-recombining region of the mating-type chromosome is associated with a unique fungal life cycle where self-fertility is enforced by maintenance of a constant state of heterokaryosis. Sequence divergence between alleles of 35 genes from the two single mating-type component strains (i.e. the homokaryotic mat A or mat a-strains), derived from one N. tetrasperma heterokaryon (mat A+mat a), was analyzed. By this approach we were able to identify the boundaries and size of the non-recombining region, and reveal insight into the history of recombination cessation. The non-recombining region covers almost 7 Mbp, over 75% of the chromosome, and we hypothesize that the evolution of the mating-type chromosome in this lineage involved two successive events. The first event was contemporaneous with the split of N. tetrasperma from a common ancestor with its outcrossing relative N. crassa and suppressed recombination over at least 6.6 Mbp, and the second was confined to a smaller region in which recombination ceased more recently. In spite of the early origin of the first "evolutionary stratum", genealogies of five genes from strains belonging to an additional N. tetrasperma lineage indicate independent initiations of suppressed recombination in different phylogenetic lineages. This study highlights the shared features between the sex chromosomes found in the animal and plant kingdoms and the fungal mating-type chromosome, despite fungi having no separate sexes. As is often found in sex chromosomes of plants and animals, recombination suppression of the mating-type chromosome of N. tetrasperma involved more than one evolutionary event, covers the majority of the mating-type chromosome and is flanked by distal regions with obligate crossovers. PMID:18369449

  10. Effect of the L499M mutation of the ascomycetous Botrytis aclada laccase on redox potential and catalytic properties

    PubMed Central

    Osipov, Evgeny; Polyakov, Konstantin; Kittl, Roman; Shleev, Sergey; Dorovatovsky, Pavel; Tikhonova, Tamara; Hann, Stephan; Ludwig, Roland; Popov, Vladimir

    2014-01-01

    Laccases are members of a large family of multicopper oxidases that catalyze the oxidation of a wide range of organic and inorganic substrates accompanied by the reduction of dioxygen to water. These enzymes contain four Cu atoms per molecule organized into three sites: T1, T2 and T3. In all laccases, the T1 copper ion is coordinated by two histidines and one cysteine in the equatorial plane and is covered by the side chains of hydrophobic residues in the axial positions. The redox potential of the T1 copper ion influences the enzymatic reaction and is determined by the nature of the axial ligands and the structure of the second coordination sphere. In this work, the laccase from the ascomycete Botrytis aclada was studied, which contains conserved Ile491 and nonconserved Leu499 residues in the axial positions. The three-dimensional structures of the wild-type enzyme and the L499M mutant were determined by X-ray crystallography at 1.7 Å resolution. Crystals suitable for X-ray analysis could only be grown after deglycosylation. Both structures did not contain the T2 copper ion. The catalytic properties of the enzyme were characterized and the redox potentials of both enzyme forms were determined: E 0 = 720 and 580 mV for the wild-type enzyme and the mutant, respectively. Since the structures of the wild-type and mutant forms are very similar, the change in the redox potential can be related to the L499M mutation in the T1 site of the enzyme. PMID:25372682

  11. Development of a Conditional Gene Expression System Using a Zearalenone-Inducible Promoter for the Ascomycete Fungus Gibberella zeae▿

    PubMed Central

    Lee, Jungkwan; Son, Hokyoung; Lee, Seunghoon; Park, Ae Ran; Lee, Yin-Won

    2010-01-01

    The ascomycete fungus Gibberella zeae is an important plant pathogen that causes fusarium head blight on small grains. Molecular studies of this fungus have been performed extensively to uncover the biological mechanisms related to pathogenicity, toxin production, and sexual reproduction. Molecular methods, such as targeted gene deletion, gene overexpression, and gene fusion to green fluorescent protein (GFP), are relatively easy to perform with this fungus; however, conditional expression systems have not been developed. The purpose of this study was to identify a promoter that could be induced by zearalenone (ZEA) for the development of a conditional expression system in G. zeae. Through microarray analysis, we isolated one zearalenone response gene (ZEAR) whose expression was increased more than 50 times after ZEA treatment. Northern blot analysis showed that the ZEAR transcript dramatically increased after 1 h of ZEA treatment. To determine the utility of the ZEAR promoter, called Pzear, in a conditional expression system, we transformed a Pzear::GFP fusion construct into G. zeae. Our data showed a ZEA concentration-dependent increase in GFP expression. We also replaced the promoter of G. zeae metE (GzmetE), an essential gene for methionine biosynthesis, with the Pzear promoter. The growth of the Pzear-GzmetE mutant on minimal medium was dependent on the ZEA concentration supplemented in the medium and showed that GzMetE expression was induced by ZEA. This study is the first report of an inducible promoter in G. zeae. Our system will be useful for the characterization of essential gene functions in this fungus through differential and ZEA-dependent gene expression. In addition, the Pzear promoter may be applicable as a biosensor for the detection of ZEA contamination in agricultural products. PMID:20348311

  12. Purifying Selection and Birth-and-Death Evolution in the Class II Hydrophobin Gene Families of the Ascomycete Trichoderma/Hypocrea

    SciTech Connect

    kubicek, Christian P.; Baker, Scott E.; Gamauf, Christian; Kenerley, Chuck; Druzhinina, Irina S.

    2008-01-10

    Hydrophobins are proteins containing eight conserved cysteine residues that occur uniquely in mycelial fungi, where their main function is to confer hydrophobicity to fungal surfaces in contact with air and during attachment of hyphae to hydrophobic surfaces of hosts, symbiotic partners or of themselves resulting in morphogenetic signals. Based on their hydropathy patterns and their solubility characteristics, they are classified in class I and class II hydrophobins, the latter being found only in ascomycetes. Here we have investigated the mechanisms driving the evolution of the class II hydrophobins in nine species of the mycoparasitic ascomycetous genus Trichoderma/Hypocrea, using three fully sequenced genomes (H. jecorina=T. reesei, H. atroviridis=T. atroviride; H. virens=T. virens) and a total of 14.000 ESTs of six others (T. asperellum, H. lixii=T. harzianum, T. aggressivum var. europeae, T. longibrachiatum, T. cf. viride). The former three contained six, ten and nine members, which is the highest number found in any other ascomycete so far. They all showed the conserved four beta-strands/one helix structure, which is stabilized by four disulfide bonds. In addition, a small number of these HFBs contained an extended N-terminus rich in either praline and aspartate, or glycine-asparagine. Phylogenetic analysis reveals a mosaic of terminal clades contain duplicated genes and shows only three reasonably supported clades. Calculation of the ratio of differences in synonymous vs. non-synonymous nucleotide substitutions provides evidence for strong purifying selection (KS/Ka >> 1). A genome database search for class II HFBs from other ascomycetes retrieved a much smaller number of hydrophobins (2-4) from each species, and most of them were from Pyrenomycetes. A combined phylogeny of these sequences with those of Trichoderma showed that the Trichoderma HFBs mostly formed their own clades, whereas those of other pyrenomycetes occured in shared clades. Our study shows that the genus Trichoderma/Hypocrea has a proliferated arsenal of class II hydrophobins which arose by purifying selection and birth-and-death evolution.

  13. A Fox2-dependent fatty acid ß-oxidation pathway coexists both in peroxisomes and mitochondria of the ascomycete yeast Candida lusitaniae.

    PubMed

    Gabriel, Frédéric; Accoceberry, Isabelle; Bessoule, Jean-Jacques; Salin, Bénédicte; Lucas-Guérin, Marine; Manon, Stephen; Dementhon, Karine; Noël, Thierry

    2014-01-01

    It is generally admitted that the ascomycete yeasts of the subphylum Saccharomycotina possess a single fatty acid ß-oxidation pathway located exclusively in peroxisomes, and that they lost mitochondrial ß-oxidation early during evolution. In this work, we showed that mutants of the opportunistic pathogenic yeast Candida lusitaniae which lack the multifunctional enzyme Fox2p, a key enzyme of the ß-oxidation pathway, were still able to grow on fatty acids as the sole carbon source, suggesting that C. lusitaniae harbored an alternative pathway for fatty acid catabolism. By assaying 14Cα-palmitoyl-CoA consumption, we demonstrated that fatty acid catabolism takes place in both peroxisomal and mitochondrial subcellular fractions. We then observed that a fox2Δ null mutant was unable to catabolize fatty acids in the mitochondrial fraction, thus indicating that the mitochondrial pathway was Fox2p-dependent. This finding was confirmed by the immunodetection of Fox2p in protein extracts obtained from purified peroxisomal and mitochondrial fractions. Finally, immunoelectron microscopy provided evidence that Fox2p was localized in both peroxisomes and mitochondria. This work constitutes the first demonstration of the existence of a Fox2p-dependent mitochondrial β-oxidation pathway in an ascomycetous yeast, C. lusitaniae. It also points to the existence of an alternative fatty acid catabolism pathway, probably located in peroxisomes, and functioning in a Fox2p-independent manner. PMID:25486052

  14. Antifungal mechanisms of a plant defensin MtDef4 are not conserved between the ascomycete fungi Neurospora crassa and Fusarium graminearum.

    PubMed

    El-Mounadi, Kaoutar; Islam, Kazi T; Hernández-Ortiz, Patricia; Read, Nick D; Shah, Dilip M

    2016-05-01

    Defensins play an important role in plant defense against fungal pathogens. The plant defensin, MtDef4, inhibits growth of the ascomycete fungi, Neurospora crassa and Fusarium graminearum, at micromolar concentrations. We have reported that MtDef4 is transported into the cytoplasm of these fungi and exerts its antifungal activity on intracellular targets. Here, we have investigated whether the antifungal mechanisms of MtDef4 are conserved in these fungi. We show that N. crassa and F. graminearum respond differently to MtDef4 challenge. Membrane permeabilization is required for the antifungal activity of MtDef4 against F. graminearum but not against N. crassa. We find that MtDef4 is targeted to different subcellular compartments in each fungus. Internalization of MtDef4 in N. crassa is energy-dependent and involves endocytosis. By contrast, MtDef4 appears to translocate into F. graminearum autonomously using a partially energy-dependent pathway. MtDef4 has been shown to bind to the phospholipid phosphatidic acid (PA). We provide evidence that the plasma membrane localized phospholipase D, involved in the biosynthesis of PA, is needed for entry of this defensin in N. crassa, but not in F. graminearum. To our knowledge, this is the first example of a defensin which inhibits the growth of two ascomycete fungi via different mechanisms. PMID:26801962

  15. Deletion of Smgpi1 encoding a GPI-anchored protein suppresses sterility of the STRIPAK mutant ?Smmob3 in the filamentous ascomycete Sordaria macrospora.

    PubMed

    Frey, Stefan; Lahmann, Yasmine; Hartmann, Thomas; Seiler, Stephan; Pggeler, Stefanie

    2015-08-01

    The striatin interacting phosphatase and kinase (STRIPAK) complex, which is composed of striatin, protein phosphatase PP2A and kinases, is required for fruiting-body development and cell fusion in the filamentous ascomycete Sordaria macrospora. Here, we report on the interplay of the glycosylphosphatidylinositol (GPI)-anchored protein SmGPI1 with the kinase activator SmMOB3, a core component of human and fungal STRIPAK complexes. SmGPI1 is conserved among filamentous ascomycetes and was first identified in a yeast two-hybrid screen using SmMOB3 as bait. The physical interaction of SmMOB3 and SmGPI1 was verified by co-immunoprecipitation. In vivo localization and differential centrifugation revealed that SmGPI1 is predominantly secreted and attached to the cell wall but is also associated with mitochondria and appears to be a dual-targeted protein. Deletion of Smgpi1 led to an increased number of fruiting bodies that were normally shaped but reduced in size. In addition, Smmob3 and Smgpi1 genetically interact. In the sterile ?Smmob3 background deletion of Smgpi1 restores fertility, vegetative growth as well as hyphal-fusion defects. The suppression effect was specific for the ?Smmob3 mutant as deletion of Smgpi1 in other STRIPAK mutants does not restore fertility. PMID:25989468

  16. Autophagy genes Smatg8 and Smatg4 are required for fruiting-body development, vegetative growth and ascospore germination in the filamentous ascomycete Sordaria macrospora.

    PubMed

    Voigt, Oliver; Pggeler, Stefanie

    2013-01-01

    Autophagy is a tightly controlled degradation process involved in various developmental aspects of eukaryotes. However, its involvement in developmental processes of multicellular filamentous ascomycetes is largely unknown. Here, we analyzed the impact of the autophagic proteins SmATG8 and SmATG4 on the sexual and vegetative development of the filamentous ascomycete Sordaria macrospora. A Saccharomyces cerevisiae complementation assay demonstrated that the S. macrospora Smatg8 and Smatg4 genes can functionally replace the yeast homologs. By generating homokaryotic deletion mutants, we showed that the S. macrospora SmATG8 and SmATG4 orthologs were associated with autophagy-dependent processes. Smatg8 and Smatg4 deletions abolished fruiting-body formation and impaired vegetative growth and ascospore germination, but not hyphal fusion. We demonstrated that SmATG4 was capable of processing the SmATG8 precursor. SmATG8 was localized to autophagosomes, whereas SmATG4 was distributed throughout the cytoplasm of S. macrospora. Furthermore, we could show that Smatg8 and Smatg4 are not only required for nonselective macroautophagy, but for selective macropexophagy as well. Taken together, our results suggest that in S. macrospora, autophagy seems to be an essential and constitutively active process to sustain high energy levels for filamentous growth and multicellular development even under nonstarvation conditions. PMID:23064313

  17. Autophagy genes Smatg8 and Smatg4 are required for fruiting-body development, vegetative growth and ascospore germination in the filamentous ascomycete Sordaria macrospora

    PubMed Central

    Voigt, Oliver; Pöggeler, Stefanie

    2013-01-01

    Autophagy is a tightly controlled degradation process involved in various developmental aspects of eukaryotes. However, its involvement in developmental processes of multicellular filamentous ascomycetes is largely unknown. Here, we analyzed the impact of the autophagic proteins SmATG8 and SmATG4 on the sexual and vegetative development of the filamentous ascomycete Sordaria macrospora. A Saccharomyces cerevisiae complementation assay demonstrated that the S. macrospora Smatg8 and Smatg4 genes can functionally replace the yeast homologs. By generating homokaryotic deletion mutants, we showed that the S. macrospora SmATG8 and SmATG4 orthologs were associated with autophagy-dependent processes. Smatg8 and Smatg4 deletions abolished fruiting-body formation and impaired vegetative growth and ascospore germination, but not hyphal fusion. We demonstrated that SmATG4 was capable of processing the SmATG8 precursor. SmATG8 was localized to autophagosomes, whereas SmATG4 was distributed throughout the cytoplasm of S. macrospora. Furthermore, we could show that Smatg8 and Smatg4 are not only required for nonselective macroautophagy, but for selective macropexophagy as well. Taken together, our results suggest that in S. macrospora, autophagy seems to be an essential and constitutively active process to sustain high energy levels for filamentous growth and multicellular development even under nonstarvation conditions. PMID:23064313

  18. Characterization of mycobionts of photomorph pairs in the peltigerineae (lichenized ascomycetes) based on internal transcribed spacer sequences of the nuclear ribosomal DNA.

    PubMed

    Goffinet, B; Bayer, R J

    1997-04-01

    The "one fungus-two photomorphs" hypothesis suggests that certain lichenized fungi can establish a symbiotic relationship with either a eukaryotic or a prokaryotic photobiont. Such pairs of photomorphs are well know from cephalodiate Peltigerineae. Using an ascomycete-specific primer we amplified the internal transcribed spacer region of the nrDNA repeat of the mycobiont from total "lichen DNA" extracts of Peltigera malacea, photomorphs of P. aphthosa, P. britannica, and P. leucophlebia, Nephroma expallidum, and photomorphs of N. arcticum. Comparisons of 5.8S sequences suggest that the sequences obtained belong to the mycobiont and thus, that the ascomycete-specific primer is adequate for amplifying fungal DNA from total lichen-DNA extracts. The strict identity of nucleotide sequences of the internal transcribed spacer region of the nrDNA repeat between joined-photomorphs supports the one fungus-two photomorphs hypothesis. Photomorph may thus primarily reflect phenotypic plasticity of photomorphic fungi in response to changing environmental conditions. The cyanomorph recently reported for P. leucophlebia is shown to be based on a misidentified specimen of P. aphthosa. Comparisons of the ITS sequences further supports recognizing P. aphthosa, P. britannica, and P. leucophlebia at the species rather than the infraspecific level. PMID:9228791

  19. Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes

    PubMed Central

    Álvarez-Cervantes, Jorge; Díaz-Godínez, Gerardo; Mercado-Flores, Yuridia; Gupta, Vijai Kumar; Anducho-Reyes, Miguel Angel

    2016-01-01

    In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to determine that the proteins under study were classified into the GH10 and GH11 families, based on the regions of highly conserved amino acids, 233–318 and 180–193 respectively, where glutamate residues are responsible for the catalysis. PMID:27040368

  20. Phylogenetic analysis of β-xylanase SRXL1 of Sporisorium reilianum and its relationship with families (GH10 and GH11) of Ascomycetes and Basidiomycetes.

    PubMed

    Álvarez-Cervantes, Jorge; Díaz-Godínez, Gerardo; Mercado-Flores, Yuridia; Gupta, Vijai Kumar; Anducho-Reyes, Miguel Angel

    2016-01-01

    In this paper, the amino acid sequence of the β-xylanase SRXL1 of Sporisorium reilianum, which is a pathogenic fungus of maize was used as a model protein to find its phylogenetic relationship with other xylanases of Ascomycetes and Basidiomycetes and the information obtained allowed to establish a hypothesis of monophyly and of biological role. 84 amino acid sequences of β-xylanase obtained from the GenBank database was used. Groupings analysis of higher-level in the Pfam database allowed to determine that the proteins under study were classified into the GH10 and GH11 families, based on the regions of highly conserved amino acids, 233-318 and 180-193 respectively, where glutamate residues are responsible for the catalysis. PMID:27040368

  1. Crystal structure of the GalNAc/Gal-specific agglutinin from the phytopathogenic ascomycete Sclerotinia sclerotiorum reveals novel adaptation of a ?-trefoil domain

    PubMed Central

    Sulzenbacher, Gerlind; Roig-Zamboni, Vronique; Peumans, Willy J.; Roug, Pierre; Van Damme, Els J.M.; Bourne, Yves

    2010-01-01

    A lectin from the phytopathogenic ascomycete Sclerotina sclerotiorum that shares only weak sequence similarity with characterized fungal lectins has recently been identified. Sclerotina sclerotiorum agglutinin (SSA) is a homodimeric protein consisting of two identical subunits of ?17 kDa and displays specificity primarily towards Gal/GalNAc. Glycan array screening indicates that SSA readily interacts with Gal/GalNAc-bearing glycan chains. The crystal structures of SSA in the ligand-free form and in complex with the Gal-?1,3-GalNAc (T-antigen) disaccharide have been determined at 1.6 and 1.97 resolution, respectively. SSA adopts a ?-trefoil domain as previously identified for other carbohydrate-binding proteins of the ricin B-like lectin superfamily and accommodates terminal non-reducing galactosyl and N-acetylgalactosaminyl glycans. Unlike other structurally related lectins, SSA contains a single carbohydrate-binding site at site ?. SSA reveals a novel dimeric assembly markedly dissimilar to those described earlier for ricin-type lectins. The present structure exemplifies the adaptability of the ?-trefoil domain in the evolution of fungal lectins. PMID:20566411

  2. Metabolites from nematophagous fungi and nematicidal natural products from fungi as an alternative for biological control. Part I: metabolites from nematophagous ascomycetes.

    PubMed

    Degenkolb, Thomas; Vilcinskas, Andreas

    2016-05-01

    Plant-parasitic nematodes are estimated to cause global annual losses of more than US$ 100 billion. The number of registered nematicides has declined substantially over the last 25 years due to concerns about their non-specific mechanisms of action and hence their potential toxicity and likelihood to cause environmental damage. Environmentally beneficial and inexpensive alternatives to chemicals, which do not affect vertebrates, crops, and other non-target organisms, are therefore urgently required. Nematophagous fungi are natural antagonists of nematode parasites, and these offer an ecophysiological source of novel biocontrol strategies. In this first section of a two-part review article, we discuss 83 nematicidal and non-nematicidal primary and secondary metabolites found in nematophagous ascomycetes. Some of these substances exhibit nematicidal activities, namely oligosporon, 4',5'-dihydrooligosporon, talathermophilins A and B, phomalactone, aurovertins D and F, paeciloxazine, a pyridine carboxylic acid derivative, and leucinostatins. Blumenol A acts as a nematode attractant. Other substances, such as arthrosporols and paganins, play a decisive role in the life cycle of the producers, regulating the formation of reproductive or trapping organs. We conclude by considering the potential applications of these beneficial organisms in plant protection strategies. PMID:26715220

  3. NPS6, Encoding a Nonribosomal Peptide Synthetase Involved in Siderophore-Mediated Iron Metabolism, Is a Conserved Virulence Determinant of Plant Pathogenic Ascomycetes[W

    PubMed Central

    Oide, Shinichi; Moeder, Wolfgang; Krasnoff, Stuart; Gibson, Donna; Haas, Hubertus; Yoshioka, Keiko; Turgeon, B. Gillian

    2006-01-01

    NPS6, encoding a nonribosomal peptide synthetase, is a virulence determinant in the maize (Zea mays) pathogen Cochliobolus heterostrophus and is involved in tolerance to H2O2. Deletion of NPS6 orthologs in the rice (Oryza sativa) pathogen, Cochliobolus miyabeanus, the wheat (Triticum aestivum) pathogen, Fusarium graminearum, and the Arabidopsis thaliana pathogen, Alternaria brassicicola, resulted in reduced virulence and hypersensitivity to H2O2. Introduction of the NPS6 ortholog from the saprobe Neurospora crassa to the Δnps6 strain of C. heterostrophus restored wild-type virulence to maize and tolerance to H2O2, demonstrating functional conservation in filamentous ascomycete phytopathogens and saprobes. Increased sensitivity to iron depletion was identified as a conserved phenotype of Δnps6 strains. Exogenous application of iron enhanced the virulence of Δnps6 strains of C. heterostrophus, C. miyabeanus, F. graminearum, and A. brassicicola to each host. NPS6 is responsible for the biosynthesis of extracellular siderophores by C. heterostrophus, F. graminearum, and A. brassicicola. Application of the extracellular siderophore of A. brassicicola restored wild-type virulence of the ΔAbnps6 strain to Arabidopsis. It is proposed that the role of extracellular siderophores in fungal virulence to plants is to supply an essential nutrient, iron, to their producers in planta and not to act as phytotoxins, depriving their hosts of iron. PMID:17056706

  4. First Structural Insights into ?-l-Arabinofuranosidases from the Two GH62 Glycoside Hydrolase Subfamilies*

    PubMed Central

    Siguier, Batrice; Haon, Mireille; Nahoum, Virginie; Marcellin, Marlne; Burlet-Schiltz, Odile; Coutinho, Pedro M.; Henrissat, Bernard; Mourey, Lionel; O'Donohue, Michael J.; Berrin, Jean-Guy; Tranier, Samuel; Dumon, Claire

    2014-01-01

    ?-l-Arabinofuranosidases are glycoside hydrolases that specifically hydrolyze non-reducing residues from arabinose-containing polysaccharides. In the case of arabinoxylans, which are the main components of hemicellulose, they are part of microbial xylanolytic systems and are necessary for complete breakdown of arabinoxylans. Glycoside hydrolase family 62 (GH62) is currently a small family of ?-l-arabinofuranosidases that contains only bacterial and fungal members. Little is known about the GH62 mechanism of action, because only a few members have been biochemically characterized and no three-dimensional structure is available. Here, we present the first crystal structures of two fungal GH62 ?-l-arabinofuranosidases from the basidiomycete Ustilago maydis (UmAbf62A) and ascomycete Podospora anserina (PaAbf62A). Both enzymes are able to efficiently remove the ?-l-arabinosyl substituents from arabinoxylan. The overall three-dimensional structure of UmAbf62A and PaAbf62A reveals a five-bladed ?-propeller fold that confirms their predicted classification into clan GH-F together with GH43 ?-l-arabinofuranosidases. Crystallographic structures of the complexes with arabinose and cellotriose reveal the important role of subsites +1 and +2 for sugar binding. Intriguingly, we observed that PaAbf62A was inhibited by cello-oligosaccharides and displayed binding affinity to cellulose although no activity was observed on a range of cellulosic substrates. Bioinformatic analyses showed that UmAbf62A and PaAbf62A belong to two distinct subfamilies within the GH62 family. The results presented here provide a framework to better investigate the structure-function relationships within the GH62 family. PMID:24394409

  5. Phylogenetic analysis of LSU and SSU rDNA group I introns of lichen photobionts associated with the genera Xanthoria and Xanthomendoza (Teloschistaceae, lichenized Ascomycetes).

    PubMed

    Nyati, Shyam; Bhattacharya, Debashish; Werth, Silke; Honegger, Rosmarie

    2013-12-01

    We studied group I introns in sterile cultures of selected groups of lichen photobionts, focusing on Trebouxia species associated with Xanthoria s. lat. (including Xanthomendoza spp.; lichen-forming ascomycetes). Group I introns were found inserted after position 798 (Escherichia coli numbering) in the large subunit (LSU) rRNA in representatives of the green algal genera Trebouxia and Asterochloris. The 798 intron was found in about 25% of Xanthoria photobionts including several reference strains obtained from algal culture collections. An alignment of LSU-encoded rDNA intron sequences revealed high similarity of these sequences allowing their phylogenetic analysis. The 798 group I intron phylogeny was largely congruent with a phylogeny of the Internal Transcribed Spacer Region (ITS), indicating that the insertion of the intron most likely occurred in the common ancestor of the genera Trebouxia and Asterochloris. The intron was vertically inherited in some taxa, but lost in others. The high sequence similarity of this intron to one found in Chlorella angustoellipsoidea suggests that the 798 intron was either present in the common ancestor of Trebouxiophyceae, or that its present distribution results from more recent horizontal transfers, followed by vertical inheritance and loss. Analysis of another group I intron shared by these photobionts at small subunit (SSU) position 1512 supports the hypothesis of repeated lateral transfers of this intron among some taxa, but loss among others. Our data confirm that the history of group I introns is characterized by repeated horizontal transfers, and suggests that some of these introns have ancient origins within Chlorophyta. PMID:24415800

  6. Biochemical and physicochemical processes contributing to the removal of endocrine-disrupting chemicals and pharmaceuticals by the aquatic ascomycete Phoma sp. UHH 5-1-03.

    PubMed

    Hofmann, Ulrike; Schlosser, Dietmar

    2016-03-01

    The environmentally widespread micropollutants bisphenol A (BPA), carbamazepine (CBZ), 17α-ethinylestradiol (EE2), diclofenac (DF), sulfamethoxazole (SMX), technical nonylphenol (t-NP) and triclosan (TCS) were used to assess the potential of the laccase-producing freshwater ascomycete Phoma sp. strain UHH 5-1-03 for micropollutant removal and to provide quantitative insights into the mechanisms involved. Biotransformation rates observed with whole fungal cells followed the rank order EE2 ≫ BPA > TCS > t-NP > DF > SMX > CBZ. Biosorption onto fungal mycelia was prominent for BPA, EE2, TCS and t-NP and insignificant for CBZ, DF and SMX. Enzymatic removal rates investigated with cell-free, laccase-containing culture supernatants of Phoma sp. followed the rank order EE2 > BPA > DF > t-NP > TCS and were insignificant for SMX and CBZ. Mass spectrometry-assisted investigations addressing metabolite formation from unlabelled and (13)C6-labelled DF and SMX yielded DF metabolites indicating hydroxylation, cyclisation and decarboxylation reactions, as well as oxidative coupling typical for laccase reactions. For SMX, several products characterised by lower molecular masses than the parent compound were found, and indications for deamination and formamide formation were obtained. Summarising, the obtained results suggest that the extracellular laccase of Phoma sp. largely contributes to fungal biotransformation of EE2, BPA, DF, TCS and t-NP, together with cell-associated enzymes such as, e.g. cytochrome P450 monooxygenases suggested by the appearance of hydroxylated metabolites from DF. Laccase does not seem to play any role in the metabolisation of SMX and CBZ, where yet to be identified cell-associated enzymes have to be considered instead. PMID:26536880

  7. Phylogenetic analysis of LSU and SSU rDNA group I introns of lichen photobionts associated with the genera Xanthoria and Xanthomendoza (Teloschistaceae, lichenized Ascomycetes)

    PubMed Central

    Nyati, Shyam; Bhattacharya, Debashish; Werth, Silke; Honegger, Rosmarie

    2013-01-01

    We studied group I introns in sterile cultures of selected groups of lichen photobionts, focusing on Trebouxia species associated with Xanthoria s. lat. (including Xanthomendoza spp.; lichen-forming ascomycetes). Group I introns were found inserted after position 798 (Escherichia coli numbering) in the large subunit (LSU) rRNA in representatives of the green algal genera Trebouxia and Asterochloris. The 798 intron was found in about 25% of Xanthoria photobionts including several reference strains obtained from algal culture collections. An alignment of LSU-encoded rDNA intron sequences revealed high similarity of these sequences allowing their phylogenetic analysis. The 798 group I intron phylogeny was largely congruent with a phylogeny of the Internal Transcribed Spacer Region (ITS), indicating that the insertion of the intron most likely occurred in the common ancestor of the genera Trebouxia and Asterochloris. The intron was vertically inherited in some taxa, but lost in others. The high sequence similarity of this intron to one found in Chlorella angustoellipsoidea suggests that the 798 intron was either present in the common ancestor of Trebouxiophyceae, or that its present distribution results from more recent horizontal transfers, followed by vertical inheritance and loss. Analysis of another group I intron shared by these photobionts at small subunit (SSU) position 1512 supports the hypothesis of repeated lateral transfers of this intron among some taxa, but loss among others. Our data confirm that the history of group I introns is characterized by repeated horizontal transfers, and suggests that some of these introns have ancient origins within Chlorophyta. PMID:24415800

  8. Minutisphaerales (Dothideomycetes, Ascomycota): a new order of freshwater ascomycetes including a new family, Minutisphaeraceae, and two new species from North Carolina, USA.

    PubMed

    Raja, Huzefa A; El-Elimat, Tamam; Oberlies, Nicholas H; Shearer, Carol A; Miller, Andrew N; Tanaka, Kazuaki; Hashimoto, Akira; Fournier, Jacques

    2015-01-01

    Minutisphaera is a recently established genus of freshwater Dothideomycetes characterized by small, globose to subglobose or apothecioid, erumpent to superficial, brown ascomata; fissitunicate, eight-spored, ovoid to obclavate asci; and 1-2-septate, clavate to broadly fusiform, hyaline to pale brown ascospores with or without a gelatinous sheath and filamentous appendages. The genus currently contains two species: M. fimbriatispora, the type species, and M. japonica. The higher-level phylogenetic relationship of Minutisphaera within the Dothideomycetes currently is unresolved. To establish the phylogenetic position of Minutisphaera within the Dothideomycetes and evaluate the phylogenetic affinities of newly collected Minutisphaera-like taxa, we sequenced three rDNA regions-18S, ITS1-5.8SITS2 (ITS) and 28S nuc rDNA, and a protein-coding gene, MCM7, for newly collected strains of Minutisphaera. Based on maximum likelihood and Bayesian analyses of a combined dataset (18S and 28S) composed of 167 taxa, a more refined dataset (28S and MCM7) comprising 52 taxa and a separate ITS dataset, and an examination of morphology, we describe and illustrate two new species of Minutisphaera. The Minutisphaera clade was strongly supported within the Dothideomycetes with likelihood and Bayesian statistics but did not share phylogenetic affinities with any existing taxonomic group within the Dothideomycetes. We therefore establish a new order, Minutisphaerales, and new family, Minutisphaeraceae, for this monophyletic clade of freshwater ascomycetes. Chemical analysis of the organic extract M. aspera (G427) resulted in isolation and characterization of five known secondary metabolites, of which four were dipeptides (1-4) and one an aromatic polyketide (5). Conversely, two aromatic polyketides (5, 6) were isolated and identified from the organic extract of M. parafimbriatispora (G156-4). The isolated compounds were tested for their antimicrobial activity against an array of bacteria and fungi. Compound 6 showed promising activity against Staphylococcus aureus and Mycobacterium smegmatis with minimal inhibitory concentration values of 30 and 60 μg/mL, respectively. PMID:26315030

  9. Real-time PCR-based monitoring of DNA pools in the tri-trophic interaction between Norway spruce, the rust Thekopsora areolata, and an opportunistic ascomycetous Phomopsis sp.

    PubMed

    Hietala, Ari M; Solheim, Halvor; Fossdal, Carl Gunnar

    2008-01-01

    The difficulty in subculturing biotrophic fungi complicates etiological studies related to the associated plant diseases. By employing internal transcribed spacer rDNA-targeted quantitative real-time polymerase chain reaction, we now show that the heteroecious rust Thekopsora areolata, commonly associated in natural conditions to sapling shoots and cones of Norway spruce and leaves of wild bird cherry, frequently infects nursery-grown seedlings of the conifer. A spatial sampling scheme was used to investigate seedlings and saplings of Norway spruce showing phloem necrosis: the highest concentration of DNA of T. areolata was recorded in the area with necrotic phloem. The separate analysis of bark and wood tissues suggested that the initial spread of the rust to healthy tissues neighboring the infection site takes place in the bark. A Phomopsis species found to coexist with T. areolata in several seedlings showed very high DNA levels in the upper part of the lesion, and even in the visually healthy proximal tissues above the lesions, which indicates that the ascomycete, most probably a secondary invader following primary infection by T. areolata, has a latent stage during early host colonization. We hypothesize that this hemibiotrophic mode of infection contributes to the successful coexistence of Phomopsis with a biotrophic rust. PMID:18943238

  10. Description of Taphrina antarctica f.a. sp. nov., a new anamorphic ascomycetous yeast species associated with Antarctic endolithic microbial communities and transfer of four Lalaria species in the genus Taphrina.

    PubMed

    Selbmann, Laura; Turchetti, Benedetta; Yurkov, Andrey; Cecchini, Clarissa; Zucconi, Laura; Isola, Daniela; Buzzini, Pietro; Onofri, Silvano

    2014-07-01

    In the framework of a large-scale rock sampling in Continental Antarctica, a number of yeasts have been isolated. Two strains that are unable to grow above 20 °C and that have low ITS sequence similarities with available data in the public domain were found. The D1/D2 LSU molecular phylogeny placed them in an isolated position in the genus Taphrina, supporting their affiliation to a not yet described species. Because the new species is able to grow in its anamorphic state only, the species Taphrina antarctica f.a. (forma asexualis) sp. nov. has been proposed to accommodate both strains (type strain DBVPG 5268(T), DSM 27485(T) and CBS 13532(T)). Lalaria and Taphrina species are dimorphic ascomycetes, where the anamorphic yeast represents the saprotrophic state and the teleomorph is the parasitic counterpart on plants. This is the first record for this genus in Antarctica; since plants are absent on the continent, we hypothesize that the fungus may have focused on the saprotrophic part of its life cycle to overcome the absence of its natural host and adapt environmental constrains. Following the new International Code of Nomenclature for Algae, Fungi and Plants (Melbourne Code 2011) the reorganization of Taphrina-Lalaria species in the teleomorphic genus Taphrina is proposed. We emend the diagnosis of the genus Taphrina to accommodate asexual saprobic states of these fungi. Taphrina antarctica was registered in MycoBank under MB 808028. PMID:24893860

  11. Pulmonary Necrotizing Granulomas in a patient with familial mediterranean fever.

    PubMed

    Kushima, Hisako; Ishii, Hiroshi; Ishii, Koji; Kadota, Jun-ichi

    2015-09-01

    We herein report a case of familial Mediterranean fever (FMF) presenting with granulomatous lung lesions with neuronal apoptosis inhibitory protein (NAIP), MHC class II transcription activator (CIITA), incompatibility locus protein from Podospora anserina (HET-E), and telomerase-associated protein (TP1) (NACHT) leucine-rich-repeat 1-positive inflammatory cell infiltrates. FMF is an autoinflammatory disorder characterized by recurrent and self-limited attacks of pyrexia, arthritis and erysipelas-like skin lesions. Lung disorders associated with FMF are extremely rare. This is the first report of an immunologically-confirmed case of pulmonary manifestations of this disease. PMID:24252001

  12. Prions are affected by evolution at two levels.

    PubMed

    Wickner, Reed B; Kelly, Amy C

    2016-03-01

    Prions, infectious proteins, can transmit diseases or be the basis of heritable traits (or both), mostly based on amyloid forms of the prion protein. A single protein sequence can be the basis for many prion strains/variants, with different biological properties based on different amyloid conformations, each rather stably propagating. Prions are unique in that evolution and selection work at both the level of the chromosomal gene encoding the protein, and on the prion itself selecting prion variants. Here, we summarize what is known about the evolution of prion proteins, both the genes and the prions themselves. We contrast the one known functional prion, [Het-s] of Podospora anserina, with the known disease prions, the yeast prions [PSI+] and [URE3] and the transmissible spongiform encephalopathies of mammals. PMID:26713322

  13. Cytotoxic Dimeric Epipolythiodiketopiperazines from the Ascomycetous Fungus Preussia typharum

    PubMed Central

    2015-01-01

    Two new dimeric epipolythiodiketopiperazines, preussiadins A (1) and B (2), together with two known diastereomers, leptosins C (6) and A (7), were obtained from the mycelia of a Preussia typharum isolate. The structures of the new compounds were established by spectroscopic methods, and the absolute configurations of 1 and 2 were assigned by chemical transformations and comparisons of quantum chemical ECD and VCD calculations to experimental data. Compound 1 exhibited potent cytotoxic activity in the NCI-60 cell line panel with an average LC50 value of 251 nM. Further studies demonstrated that 1 circumvents P-glycoprotein-mediated drug resistance, yet had no significant antitumor activity in a xenograft UACC-62 melanoma model. PMID:24893224

  14. Fatal cerebral mycoses caused by the ascomycete Chaetomium strumarium.

    PubMed Central

    Abbott, S P; Sigler, L; McAleer, R; McGough, D A; Rinaldi, M G; Mizell, G

    1995-01-01

    Three cases of fatal cerebral mycosis in males with prior histories of intravenous drug use from the United States and Australia are reported. Infection in each case was limited to brain abscess; no other sites of infection were observed. The fungus seen by histopathology and isolated from the brain tissue in each case was identified as Chaetomium strumarium. This is the first report of human infection by this species, and C. strumarium is the second species of Chaetomium known to cause primary brain infection. Chaetomium strumarium is unusual among members of the genus Chaetomium in forming ascocarps covered with pale, thin-walled, flexuous hairs, a feature leading to its original placement in the genus Achaetomium. Presence of pinkish exudate droplets and/or crystals associated with hyphae or ascocarps, sometimes accompanied by a pinkish diffusible pigment; good growth at 42 degrees C; and production of small conidia further distinguish this species. The brain abscess isolates were compared with isolates from prior cases of cerebral infection which had been identified as either Chaetomium atrobrunneum or Chaetomium globosum. With reidentification of one isolate originally identified as C. globosum to C. atrobrunneum, only C. strumarium and C. atrobrunneum have been confirmed to cause infection involving the brain. PMID:8567907

  15. Enzymes and bioproducts produced by the ascomycete fungus Paecilomyces variotii.

    PubMed

    Herrera Bravo de Laguna, I; Toledo Marante, F J; Mioso, R

    2015-12-01

    Due its innate ability to produce extracellular enzymes which can provide eco-friendly solutions for a variety of biotechnological applications, Paecilomyces variotii is a potential source of industrial bioproducts. In this review, we report biotechnological records on the biochemistry of different enzymes produced by the fermentation of the P. variotii fungus, including tannases, phytases, cellulases, xylanases, chitinases, amylases and pectinases. Additionally, the main physicochemical properties which can affect the enzymatic reactions of the enzymes involved in the conversion of a huge number of substrates to high-value bioproducts are described. Despite all the background information compiled in this review, more research is required to consolidate the catalytic efficiency of P. variotii, which must be optimized so that it is more accurate and reproducible on a large scale. PMID:26274842

  16. Nuclear and Genome Dynamics in Multinucleate Ascomycete Fungi

    PubMed Central

    Roper, Marcus; Ellison, Chris; Taylor, John W.; Glass, N. Louise

    2011-01-01

    Genetic variation between individuals is essential to evolution and adaptation. However, intra-organismic genetic variation also shapes the life histories of many organisms, including filamentous fungi. A single fungal syncytium can harbor thousands or millions of mobile and potentially genotypically different nuclei, each having the capacity to regenerate a new organism. Because the dispersal of asexual or sexual spores propagates individual nuclei in many of these species, selection acting at the level of nuclei creates the potential for competitive and cooperative genome dynamics. Recent work in Neurospora crassa and Sclerotinia sclerotiorum has illuminated how nuclear populations are coordinated for fungal growth and other behaviors and has revealed both molecular and physical mechanisms for preventing and policing inter-genomic conflict. Recent results from population-level genomic studies in a variety of filamentous fungi suggest that nuclear exchange between mycelia and recombination between heterospecific nuclei may be of more importance to fungal evolution, diversity and the emergence of newly virulent strains than has previously been recognized. PMID:21959169

  17. Characterization of the aodA, dnmA, mnSOD and pimA genes in Aspergillus nidulans.

    PubMed

    Leiter, Éva; Park, Hee-Soo; Kwon, Nak-Jung; Han, Kap-Hoon; Emri, Tamás; Oláh, Viktor; Mészáros, Ilona; Dienes, Beatrix; Vincze, János; Csernoch, László; Yu, Jae-Hyuk; Pócsi, István

    2016-01-01

    Mitochondria play key roles in cellular energy generation and lifespan of most eukaryotes. To understand the functions of four nuclear-encoded genes predicted to be related to the maintenance of mitochondrial morphology and function in Aspergillus nidulans, systematic characterization was carried out. The deletion and overexpression mutants of aodA, dnmA, mnSOD and pimA encoding alternative oxidase, dynamin related protein, manganese superoxide dismutase and Lon protease, respectively, were generated and examined for their growth, stress tolerances, respiration, autolysis, cell death, sterigmatocystin production, hyphal morphology and size, and mitochondrial superoxide production as well as development. Overall, genetic manipulation of these genes had less effect on cellular physiology and ageing in A. nidulans than that of their homologs in another fungus Podospora anserina with a well-characterized senescence. The observed interspecial phenotypic differences can be explained by the dissimilar intrinsic stabilities of the mitochondrial genomes in A. nidulans and P. anserina. Furthermore, the marginally altered phenotypes observed in A. nidulans mutants indicate the presence of effective compensatory mechanisms for the complex networks of mitochondrial defense and quality control. Importantly, these findings can be useful for developing novel platforms for heterologous protein production, or on new biocontrol and bioremediation technologies based on Aspergillus species. PMID:26846452

  18. A mitochondrial mutator plasmid that causes senescence under dietary restricted conditions

    PubMed Central

    Maas, Marc FPM; Hoekstra, Rolf F; Debets, Alfons JM

    2007-01-01

    Background Calorie or dietary restriction extends life span in a wide range of organisms including the filamentous fungus Podospora anserina. Under dietary restricted conditions, P. anserina isolates are several-fold longer lived. This is however not the case in isolates that carry one of the pAL2-1 homologous mitochondrial plasmids. Results We show that the pAL2-1 homologues act as 'insertional mutators' of the mitochondrial genome, which may explain their negative effect on life span extension. Sequencing revealed at least fourteen unique plasmid integration sites, of which twelve were located within the mitochondrial genome and two within copies of the plasmid itself. The plasmids were able to integrate in their entirety, via a non-homologous mode of recombination. Some of the integrated plasmid copies were truncated, which probably resulted from secondary, post-integrative, recombination processes. Integration sites were predominantly located within and surrounding the region containing the mitochondrial rDNA loci. Conclusion We propose a model for the mechanism of integration, based on innate modes of mtDNA recombination, and discuss its possible link with the plasmid's negative effect on dietary restriction mediated life span extension. PMID:17407571

  19. Characterization of the aodA, dnmA, mnSOD and pimA genes in Aspergillus nidulans

    PubMed Central

    Leiter, Éva; Park, Hee-Soo; Kwon, Nak-Jung; Han, Kap-Hoon; Emri, Tamás; Oláh, Viktor; Mészáros, Ilona; Dienes, Beatrix; Vincze, János; Csernoch, László; Yu, Jae-Hyuk; Pócsi, István

    2016-01-01

    Mitochondria play key roles in cellular energy generation and lifespan of most eukaryotes. To understand the functions of four nuclear-encoded genes predicted to be related to the maintenance of mitochondrial morphology and function in Aspergillus nidulans, systematic characterization was carried out. The deletion and overexpression mutants of aodA, dnmA, mnSOD and pimA encoding alternative oxidase, dynamin related protein, manganese superoxide dismutase and Lon protease, respectively, were generated and examined for their growth, stress tolerances, respiration, autolysis, cell death, sterigmatocystin production, hyphal morphology and size, and mitochondrial superoxide production as well as development. Overall, genetic manipulation of these genes had less effect on cellular physiology and ageing in A. nidulans than that of their homologs in another fungus Podospora anserina with a well-characterized senescence. The observed interspecial phenotypic differences can be explained by the dissimilar intrinsic stabilities of the mitochondrial genomes in A. nidulans and P. anserina. Furthermore, the marginally altered phenotypes observed in A. nidulans mutants indicate the presence of effective compensatory mechanisms for the complex networks of mitochondrial defense and quality control. Importantly, these findings can be useful for developing novel platforms for heterologous protein production, or on new biocontrol and bioremediation technologies based on Aspergillus species. PMID:26846452

  20. An Acetyltransferase Conferring Tolerance to Toxic Aromatic Amine Chemicals

    PubMed Central

    Martins, Marta; Rodrigues-Lima, Fernando; Dairou, Julien; Lamouri, Aazdine; Malagnac, Fabienne; Silar, Philippe; Dupret, Jean-Marie

    2009-01-01

    Aromatic amines (AA) are a major class of environmental pollutants that have been shown to have genotoxic and cytotoxic potentials toward most living organisms. Fungi are able to tolerate a diverse range of chemical compounds including certain AA and have long been used as models to understand general biological processes. Deciphering the mechanisms underlying this tolerance may improve our understanding of the adaptation of organisms to stressful environments and pave the way for novel pharmaceutical and/or biotechnological applications. We have identified and characterized two arylamine N-acetyltransferase (NAT) enzymes (PaNAT1 and PaNAT2) from the model fungus Podospora anserina that acetylate a wide range of AA. Targeted gene disruption experiments revealed that PaNAT2 was required for the growth and survival of the fungus in the presence of toxic AA. Functional studies using the knock-out strains and chemically acetylated AA indicated that tolerance of P. anserina to toxic AA was due to the N-acetylation of these chemicals by PaNAT2. Moreover, we provide proof-of-concept remediation experiments where P. anserina, through its PaNAT2 enzyme, is able to detoxify the highly toxic pesticide residue 3,4-dichloroaniline in experimentally contaminated soil samples. Overall, our data show that a single xenobiotic-metabolizing enzyme can mediate tolerance to a major class of pollutants in a eukaryotic species. These findings expand the understanding of the role of xenobiotic-metabolizing enzyme and in particular of NATs in the adaptation of organisms to their chemical environment and provide a basis for new systems for the bioremediation of contaminated soils. PMID:19416981

  1. An acetyltransferase conferring tolerance to toxic aromatic amine chemicals: molecular and functional studies.

    PubMed

    Martins, Marta; Rodrigues-Lima, Fernando; Dairou, Julien; Lamouri, Aazdine; Malagnac, Fabienne; Silar, Philippe; Dupret, Jean-Marie

    2009-07-10

    Aromatic amines (AA) are a major class of environmental pollutants that have been shown to have genotoxic and cytotoxic potentials toward most living organisms. Fungi are able to tolerate a diverse range of chemical compounds including certain AA and have long been used as models to understand general biological processes. Deciphering the mechanisms underlying this tolerance may improve our understanding of the adaptation of organisms to stressful environments and pave the way for novel pharmaceutical and/or biotechnological applications. We have identified and characterized two arylamine N-acetyltransferase (NAT) enzymes (PaNAT1 and PaNAT2) from the model fungus Podospora anserina that acetylate a wide range of AA. Targeted gene disruption experiments revealed that PaNAT2 was required for the growth and survival of the fungus in the presence of toxic AA. Functional studies using the knock-out strains and chemically acetylated AA indicated that tolerance of P. anserina to toxic AA was due to the N-acetylation of these chemicals by PaNAT2. Moreover, we provide proof-of-concept remediation experiments where P. anserina, through its PaNAT2 enzyme, is able to detoxify the highly toxic pesticide residue 3,4-dichloroaniline in experimentally contaminated soil samples. Overall, our data show that a single xenobiotic-metabolizing enzyme can mediate tolerance to a major class of pollutants in a eukaryotic species. These findings expand the understanding of the role of xenobiotic-metabolizing enzyme and in particular of NATs in the adaptation of organisms to their chemical environment and provide a basis for new systems for the bioremediation of contaminated soils. PMID:19416981

  2. Global Protein Oxidation Profiling Suggests Efficient Mitochondrial Proteome Homeostasis During Aging.

    PubMed

    Ramallo Guevara, Carina; Philipp, Oliver; Hamann, Andrea; Werner, Alexandra; Osiewacz, Heinz D; Rexroth, Sascha; Rögner, Matthias; Poetsch, Ansgar

    2016-05-01

    The free radical theory of aging is based on the idea that reactive oxygen species (ROS) may lead to the accumulation of age-related protein oxidation. Because themajority of cellular ROS is generated at the respiratory electron transport chain, this study focuses on the mitochondrial proteome of the aging model Podospora anserina as target for ROS-induced damage. To ensure the detection of even low abundant modified peptides, separation by long gradient nLC-ESI-MS/MS and an appropriate statistical workflow for iTRAQ quantification was developed. Artificial protein oxidation was minimized by establishing gel-free sample preparation in the presence of reducing and iron-chelating agents. This first large scale, oxidative modification-centric study for P. anserina allowed the comprehensive quantification of 22 different oxidative amino acid modifications, and notably the quantitative comparison of oxidized and nonoxidized protein species. In total 2341 proteins were quantified. For 746 both protein species (unmodified and oxidatively modified) were detected and the modification sites determined. The data revealed that methionine residues are preferably oxidized. Further prominent identified modifications in decreasing order of occurrence were carbonylation as well as formation of N-formylkynurenine and pyrrolidinone. Interestingly, for the majority of proteins a positive correlation of changes in protein amount and oxidative damage were noticed, and a general decrease in protein amounts at late age. However, it was discovered that few proteins changed in oxidative damage in accordance with former reports. Our data suggest that P. anserina is efficiently capable to counteract ROS-induced protein damage during aging as long as protein de novo synthesis is functioning, ultimately leading to an overall constant relationship between damaged and undamaged protein species. These findings contradict a massive increase in protein oxidation during aging and rather suggest a protein damage homeostasis mechanism even at late age. PMID:26884511

  3. The NADPH Oxidase Complexes in Botrytis cinerea: Evidence for a Close Association with the ER and the Tetraspanin Pls1

    PubMed Central

    Siegmund, Ulrike; Heller, Jens; van Kann, Jan A. L.; Tudzynski, Paul

    2013-01-01

    NADPH oxidases (Nox) are major enzymatic systems that generate reactive-oxygen species (ROS) in multicellular eukaryotes. In several fungi they have been shown to be involved in sexual differentiation and pathogenicity. However, in contrast to the well characterized mammalian systems, basic information on the composition, recruitment, and localization of fungal Nox complexes and on the molecular mechanisms of their cellular effects are still lacking. Here we give a detailed analysis of components of the Nox complexes in the gray mold fungus Botrytis cinerea. It had previously been shown that the two catalytic transmembrane subunits BcNoxA and B are important for development of sclerotia and for full virulence, with BcNoxA being involved in spreading of lesions and BcNoxB in penetration; BcNoxR functions as a regulator of both subunits. Here we present evidence (using for the first time a functional GFP fusion able to complement the ΔbcnoxA mutant) that BcNoxA localizes mainly to the ER and at the plasma membrane; BcNoxB shows a similar localization pattern, while the regulator BcNoxR is found in vesicles throughout the hyphae and at the hyphal tip. To identify possible interaction partners, which could be involved in the localization or recruitment of the Nox complexes, we functionally characterized the tetraspanin Pls1, a transmembrane protein, which had been suggested to be a NoxB-interacting partner in the saprophyte Podospora anserina. Knock-out experiments and GFP fusions substantiate a link between BcNoxB and BcPls1 because both deletion mutants have overlapping phenotypes (especially a defect in penetration), and the proteins show a similar localization pattern (ER). However, in contrast to the corresponding protein in P. anserina BcPls1 is important for female fertility, but not for ascospore germination. PMID:23418468

  4. The NADPH oxidase complexes in Botrytis cinerea: evidence for a close association with the ER and the tetraspanin Pls1.

    PubMed

    Siegmund, Ulrike; Heller, Jens; van Kan, Jan A L; van Kann, Jan A L; Tudzynski, Paul

    2013-01-01

    NADPH oxidases (Nox) are major enzymatic systems that generate reactive-oxygen species (ROS) in multicellular eukaryotes. In several fungi they have been shown to be involved in sexual differentiation and pathogenicity. However, in contrast to the well characterized mammalian systems, basic information on the composition, recruitment, and localization of fungal Nox complexes and on the molecular mechanisms of their cellular effects are still lacking. Here we give a detailed analysis of components of the Nox complexes in the gray mold fungus Botrytis cinerea. It had previously been shown that the two catalytic transmembrane subunits BcNoxA and B are important for development of sclerotia and for full virulence, with BcNoxA being involved in spreading of lesions and BcNoxB in penetration; BcNoxR functions as a regulator of both subunits. Here we present evidence (using for the first time a functional GFP fusion able to complement the ?bcnoxA mutant) that BcNoxA localizes mainly to the ER and at the plasma membrane; BcNoxB shows a similar localization pattern, while the regulator BcNoxR is found in vesicles throughout the hyphae and at the hyphal tip. To identify possible interaction partners, which could be involved in the localization or recruitment of the Nox complexes, we functionally characterized the tetraspanin Pls1, a transmembrane protein, which had been suggested to be a NoxB-interacting partner in the saprophyte Podospora anserina. Knock-out experiments and GFP fusions substantiate a link between BcNoxB and BcPls1 because both deletion mutants have overlapping phenotypes (especially a defect in penetration), and the proteins show a similar localization pattern (ER). However, in contrast to the corresponding protein in P. anserina BcPls1 is important for female fertility, but not for ascospore germination. PMID:23418468

  5. Peroxisomes and sexual development in fungi

    PubMed Central

    Peraza-Reyes, Leonardo; Berteaux-Lecellier, Véronique

    2013-01-01

    Peroxisomes are versatile and dynamic organelles that are essential for the development of most eukaryotic organisms. In fungi, many developmental processes, such as sexual development, require the activity of peroxisomes. Sexual reproduction in fungi involves the formation of meiotic-derived sexual spores, often takes place inside multicellular fruiting bodies and requires precise coordination between the differentiation of multiple cell types and the progression of karyogamy and meiosis. Different peroxisomal functions contribute to the orchestration of this complex developmental process. Peroxisomes are required to sustain the formation of fruiting bodies and the maturation and germination of sexual spores. They facilitate the mobilization of reserve compounds via fatty acid β-oxidation and the glyoxylate cycle, allowing the generation of energy and biosynthetic precursors. Additionally, peroxisomes are implicated in the progression of meiotic development. During meiotic development in Podospora anserina, there is a precise modulation of peroxisome assembly and dynamics. This modulation includes changes in peroxisome size, number and localization, and involves a differential activity of the protein-machinery that drives the import of proteins into peroxisomes. Furthermore, karyogamy, entry into meiosis and sorting of meiotic-derived nuclei into sexual spores all require the activity of peroxisomes. These processes rely on different peroxisomal functions and likely depend on different pathways for peroxisome assembly. Indeed, emerging studies support the existence of distinct import channels for peroxisomal proteins that contribute to different developmental stages. PMID:24046747

  6. [Yeast prions as a model of neurodegenerative infectious amyloidoses in humans].

    PubMed

    Inge-Vechtomov, S G

    2011-01-01

    Several neurodegenerative diseases (so-called age-related diseases) in humans are associated with development of protein aggregates--amyloids. Prion diseases--kuru, Kreutzfeldt-Jakob and Gerstmann-Straussler-Sheinker diseases, fatal familial insomnia, etc.--are examples of infectious amyloidoses. A model system for investigation of mechanisms of amyloidogenesis and of its infectious nature had been developed as a result of yeast prion discovery. The existence of a prion network as an interaction of different prions identified in yeast is being confirmed recently as an interaction of different anyloids in humans. The potential danger of amyloidoses is conditioned by the very structure of almost all proteins containing fragments capable to be organized as beta-sheets, which lead to their aggregation being exposed. Meanwhile, there are several well-defined examples of the adaptive value of amyloid aggregates: cytoplasmic incompatibility factor in Podospora anserina, spider silk, cytoplasmic stress granules in mammals, prion form of CPEB protein responsible for the neuron activity in Aplisia, etc. These facts should be taken into consideration when seeking antiamyloid drugs. Discovery of protein inheritance in lower eukaryotes modifies our knowledge of the template principle significance in biology and adds a concept of conformational templates (II order templates) involved in reproduction of the three-dimensional structure of the supramolecular complexes in the cell. PMID:22145302

  7. A mitotically inheritable unit containing a MAP kinase module

    PubMed Central

    Kicka, Sébastien; Bonnet, Crystel; Sobering, Andrew K.; Ganesan, Latha P.; Silar, Philippe

    2006-01-01

    Prions are novel kinds of hereditary units, relying solely on proteins, that are infectious and inherited in a non-Mendelian fashion. To date, they are either based on autocatalytic modification of a 3D conformation or on autocatalytic cleavage. Here, we provide further evidence that in the filamentous fungus Podospora anserina, a MAP kinase cascade is probably able to self-activate and generate C, a hereditary unit that bears many similarities to prions and triggers cell degeneration. We show that in addition to the MAPKKK gene, both the MAPKK and MAPK genes are necessary for the propagation of C, and that overexpression of MAPK as that of MAPKKK facilitates the appearance of C. We also show that a correlation exists between the presence of C and localization of the MAPK inside nuclei. These data emphasize the resemblance between prions and a self-positively regulated cascade in terms of their transmission. This thus further expands the concept of protein-base inheritance to regulatory networks that have the ability to self-activate. PMID:16938837

  8. Theme and variations: evolutionary diversification of the HET-s functional amyloid motif

    PubMed Central

    Daskalov, Asen; Dyrka, Witold; Saupe, Sven J.

    2015-01-01

    In mammals and fungi, Nod-like receptors (NLR) activate downstream cell death execution proteins by a prion-like mechanism. In Podospora anserina, the NWD2 NLR activates the HET-S Helo-domain pore-forming protein by converting its prion-forming domain into a characteristic ?-solenoid amyloid fold. The amyloid forming region of HET-S/s comprises two repetitions of a 21 amino acid motif. Herein, we systematically analyze the sequences of C-terminal regions of fungal HeLo and HeLo-like domain proteins to identify HET-s-related amyloid motifs (HRAM). We now identify four novel HRAM subfamilies in addition to the canonical HET-S/s subfamily. These novel motifs share the pseudo-repeat structure of HET-S/s and a specific pattern of distribution of hydrophobic and polar residues. Sequence co-variance analyses predict parallel in-register ?-stacking of the two repeats and residue-residue interactions compatible with the ?-solenoid fold. As described for HET-S, most genes encoding the HeLo proteins are adjacent to genes encoding NLRs also displaying HRAMs. The motifs of the NLRs are similar to those of their cognate HeLo-domain protein, indicating concerted evolution between repeats. This study shows that HET-s-related amyloid motifs are more common than anticipated and that they have diversified into discrete subfamilies that apparently share a common overall fold. PMID:26219477

  9. Yeast and Fungal Prions: Amyloid-Handling Systems, Amyloid Structure, and Prion Biology.

    PubMed

    Wickner, R B; Edskes, H K; Gorkovskiy, A; Bezsonov, E E; Stroobant, E E

    2016-01-01

    Yeast prions (infectious proteins) were discovered by their outré genetic properties and have become important models for an array of human prion and amyloid diseases. A single prion protein can become any of many distinct amyloid forms (called prion variants or strains), each of which is self-propagating, but with different biological properties (eg, lethal vs mild). The folded in-register parallel β sheet architecture of the yeast prion amyloids naturally suggests a mechanism by which prion variant information can be faithfully transmitted for many generations. The yeast prions rely on cellular chaperones for their propagation, but can be cured by various chaperone imbalances. The Btn2/Cur1 system normally cures most variants of the [URE3] prion that arise. Although most variants of the [PSI+] and [URE3] prions are toxic or lethal, some are mild in their effects. Even the most mild forms of these prions are rare in the wild, indicating that they too are detrimental to yeast. The beneficial [Het-s] prion of Podospora anserina poses an important contrast in its structure, biology, and evolution to the yeast prions characterized thus far. PMID:26915272

  10. Molecular Characterization of a New Alkaline-Tolerant Xylanase from Humicola insolens Y1

    PubMed Central

    Shi, Pengjun; Du, Yanlong; Yang, Hong; Huang, Huoqing; Zhang, Xiu; Wang, Yaru; Yao, Bin

    2015-01-01

    An endo-1,4-β-xylanase-encoding gene, xyn11B, was cloned from the thermophilic fungus Humicola insolens Y1. The gene encodes a multimodular xylanase that consists of a typical hydrophobic signal sequence, a catalytic domain of glycoside hydrolase (GH) family 11, a glycine-rich linker, and a family 1 carbohydrate binding module (CBM1). Deduced Xyn11B shares the highest identity of 74% with a putative xylanase from Podospora anserina S mat+. Recombinant Xyn11B was successfully expressed in Pichia pastoris and purified to electrophoretic homogeneity. Xyn11B had a high specific activity of 382.0 U mg−1 towards beechwood xylan and showed optimal activity at pH 6.0 and 50°C. Distinct from most reported acidic fungal xylanases, Xyn11B was alkaline-tolerant, retaining 30.7% of the maximal activity at pH 9.0. The Km and Vmax values for beechwood xylan were 2.2 mg mL−1 and 462.8 μmol min−1 mg−1, respectively. The enzyme exhibited a wider substrate specificity and produced a mixture of xylooligosaccharides. All these favorable enzymatic properties make Xyn11B attractive for potential applications in various industries. PMID:25629035

  11. Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism

    PubMed Central

    Fischer, Fabian; Langer, Julian D.; Osiewacz, Heinz D.

    2015-01-01

    Maintenance of mitochondria is achieved by several mechanisms, including the regulation of mitochondrial proteostasis. The matrix protease CLPXP, involved in protein quality control, has been implicated in ageing and disease. However, particularly due to the lack of knowledge of CLPXP’s substrate spectrum, only little is known about the pathways and mechanisms controlled by this protease. Here we report the first comprehensive identification of potential mitochondrial CLPXP in vivo interaction partners and substrates using a combination of tandem affinity purification and differential proteomics. This analysis reveals that CLPXP in the fungal ageing model Podospora anserina is mainly associated with metabolic pathways in mitochondria, e.g. components of the pyruvate dehydrogenase complex and the tricarboxylic acid cycle as well as subunits of electron transport chain complex I. These data suggest a possible function of mitochondrial CLPXP in the control and/or maintenance of energy metabolism. Since bioenergetic alterations are a common feature of neurodegenerative diseases, cancer, and ageing, our data comprise an important resource for specific studies addressing the role of CLPXP in these adverse processes. PMID:26679294

  12. Theme and variations: evolutionary diversification of the HET-s functional amyloid motif.

    PubMed

    Daskalov, Asen; Dyrka, Witold; Saupe, Sven J

    2015-01-01

    In mammals and fungi, Nod-like receptors (NLR) activate downstream cell death execution proteins by a prion-like mechanism. In Podospora anserina, the NWD2 NLR activates the HET-S Helo-domain pore-forming protein by converting its prion-forming domain into a characteristic ?-solenoid amyloid fold. The amyloid forming region of HET-S/s comprises two repetitions of a 21 amino acid motif. Herein, we systematically analyze the sequences of C-terminal regions of fungal HeLo and HeLo-like domain proteins to identify HET-s-related amyloid motifs (HRAM). We now identify four novel HRAM subfamilies in addition to the canonical HET-S/s subfamily. These novel motifs share the pseudo-repeat structure of HET-S/s and a specific pattern of distribution of hydrophobic and polar residues. Sequence co-variance analyses predict parallel in-register ?-stacking of the two repeats and residue-residue interactions compatible with the ?-solenoid fold. As described for HET-S, most genes encoding the HeLo proteins are adjacent to genes encoding NLRs also displaying HRAMs. The motifs of the NLRs are similar to those of their cognate HeLo-domain protein, indicating concerted evolution between repeats. This study shows that HET-s-related amyloid motifs are more common than anticipated and that they have diversified into discrete subfamilies that apparently share a common overall fold. PMID:26219477

  13. Biotransformation of Trichoderma spp. and their tolerance to aromatic amines, a major class of pollutants.

    PubMed

    Cocaign, Angélique; Bui, Linh-Chi; Silar, Philippe; Chan Ho Tong, Laetitia; Busi, Florent; Lamouri, Aazdine; Mougin, Christian; Rodrigues-Lima, Fernando; Dupret, Jean-Marie; Dairou, Julien

    2013-08-01

    Trichoderma spp. are cosmopolitan soil fungi that are highly resistant to many toxic compounds. Here, we show that Trichoderma virens and T. reesei are tolerant to aromatic amines (AA), a major class of pollutants including the highly toxic pesticide residue 3,4-dichloroaniline (3,4-DCA). In a previous study, we provided proof-of-concept remediation experiments in which another soil fungus, Podospora anserina, detoxifies 3,4-DCA through its arylamine N-acetyltransferase (NAT), a xenobiotic-metabolizing enzyme that enables acetyl coenzyme A-dependent detoxification of AA. To assess whether the N-acetylation pathway enables AA tolerance in Trichoderma spp., we cloned and characterized NATs from T. virens and T. reesei. We characterized recombinant enzymes by determining their catalytic efficiencies toward several toxic AA. Through a complementary approach, we also demonstrate that both Trichoderma species efficiently metabolize 3,4-DCA. Finally, we provide evidence that NAT-independent transformation is solely (in T. virens) or mainly (in T. reesei) responsible for the observed removal of 3,4-DCA. We conclude that T. virens and, to a lesser extent, T. reesei likely utilize another, unidentified, metabolic pathway for the detoxification of AA aside from acetylation. This is the first molecular and functional characterization of AA biotransformation in Trichoderma spp. Given the potential of Trichoderma for cleanup of contaminated soils, these results reveal new possibilities in the fungal remediation of AA-contaminated soil. PMID:23728813

  14. Biotransformation of Trichoderma spp. and Their Tolerance to Aromatic Amines, a Major Class of Pollutants

    PubMed Central

    Cocaign, Angélique; Bui, Linh-Chi; Silar, Philippe; Chan Ho Tong, Laetitia; Busi, Florent; Lamouri, Aazdine; Mougin, Christian; Rodrigues-Lima, Fernando

    2013-01-01

    Trichoderma spp. are cosmopolitan soil fungi that are highly resistant to many toxic compounds. Here, we show that Trichoderma virens and T. reesei are tolerant to aromatic amines (AA), a major class of pollutants including the highly toxic pesticide residue 3,4-dichloroaniline (3,4-DCA). In a previous study, we provided proof-of-concept remediation experiments in which another soil fungus, Podospora anserina, detoxifies 3,4-DCA through its arylamine N-acetyltransferase (NAT), a xenobiotic-metabolizing enzyme that enables acetyl coenzyme A-dependent detoxification of AA. To assess whether the N-acetylation pathway enables AA tolerance in Trichoderma spp., we cloned and characterized NATs from T. virens and T. reesei. We characterized recombinant enzymes by determining their catalytic efficiencies toward several toxic AA. Through a complementary approach, we also demonstrate that both Trichoderma species efficiently metabolize 3,4-DCA. Finally, we provide evidence that NAT-independent transformation is solely (in T. virens) or mainly (in T. reesei) responsible for the observed removal of 3,4-DCA. We conclude that T. virens and, to a lesser extent, T. reesei likely utilize another, unidentified, metabolic pathway for the detoxification of AA aside from acetylation. This is the first molecular and functional characterization of AA biotransformation in Trichoderma spp. Given the potential of Trichoderma for cleanup of contaminated soils, these results reveal new possibilities in the fungal remediation of AA-contaminated soil. PMID:23728813

  15. MitoBreak: the mitochondrial DNA breakpoints database

    PubMed Central

    Damas, Joana; Carneiro, João; Amorim, António; Pereira, Filipe

    2014-01-01

    Mitochondrial DNA (mtDNA) rearrangements are key events in the development of many diseases. Investigations of mtDNA regions affected by rearrangements (i.e. breakpoints) can lead to important discoveries about rearrangement mechanisms and can offer important clues about the causes of mitochondrial diseases. Here, we present the mitochondrial DNA breakpoints database (MitoBreak; http://mitobreak.portugene.com), a free, web-accessible comprehensive list of breakpoints from three classes of somatic mtDNA rearrangements: circular deleted (deletions), circular partially duplicated (duplications) and linear mtDNAs. Currently, MitoBreak contains >1400 mtDNA rearrangements from seven species (Homo sapiens, Mus musculus, Rattus norvegicus, Macaca mulatta, Drosophila melanogaster, Caenorhabditis elegans and Podospora anserina) and their associated phenotypic information collected from nearly 400 publications. The database allows researchers to perform multiple types of data analyses through user-friendly interfaces with full or partial datasets. It also permits the download of curated data and the submission of new mtDNA rearrangements. For each reported case, MitoBreak also documents the precise breakpoint positions, junction sequences, disease or associated symptoms and links to the related publications, providing a useful resource to study the causes and consequences of mtDNA structural alterations. PMID:24170808

  16. RCF1-dependent respiratory supercomplexes are integral for lifespan-maintenance in a fungal ageing model

    PubMed Central

    Fischer, Fabian; Filippis, Christodoulos; Osiewacz, Heinz D.

    2015-01-01

    Mitochondrial respiratory supercomplexes (mtRSCs) are stoichiometric assemblies of electron transport chain (ETC) complexes in the inner mitochondrial membrane. They are hypothesized to regulate electron flow, the generation of reactive oxygen species (ROS) and to stabilize ETC complexes. Using the fungal ageing model Podospora anserina, we investigated the impact of homologues of the Saccharomyces cerevisiae respiratory supercomplex factors 1 and 2 (termed PaRCF1 and PaRCF2) on mtRSC formation, fitness and lifespan. Whereas PaRCF2’s role seems negligible, ablation of PaRCF1 alters size of monomeric complex IV, reduces the abundance of complex IV-containing supercomplexes, negatively affects vital functions and shortens lifespan. PaRcf1 overexpression slightly prolongs lifespan, though without appreciably influencing ETC organization. Overall, our results identify PaRCF1 as necessary yet not sufficient for mtRSC formation and demonstrate that PaRCF1-dependent stability of complex IV and associated supercomplexes is highly relevant for maintenance of the healthy lifespan in a eukaryotic model organism. PMID:26220011

  17. A Putative Transcription Factor MYT1 Is Required for Female Fertility in the Ascomycete Gibberella zeae

    PubMed Central

    Lin, Yang; Son, Hokyoung; Lee, Jungkwan; Min, Kyunghun; Choi, Gyung Ja; Kim, Jin-Cheol; Lee, Yin-Won

    2011-01-01

    Gibberella zeae is an important pathogen of major cereal crops. The fungus produces ascospores that forcibly discharge from mature fruiting bodies, which serve as the primary inocula for disease epidemics. In this study, we characterized an insertional mutant Z39P105 with a defect in sexual development and identified a gene encoding a putative transcription factor designated as MYT1. This gene contains a Myb DNA-binding domain and is conserved in the subphylum Pezizomycotina of Ascomycota. The MYT1 protein fused with green fluorescence protein localized in nuclei, which supports its role as a transcriptional regulator. The MYT1 deletion mutant showed similar phenotypes to the wild-type strain in vegetative growth, conidia production and germination, virulence, and mycotoxin production, but had defect in female fertility. A mutant overexpressing MYT1 showed earlier germination, faster mycelia growth, and reduced mycotoxin production compared to the wild-type strain, suggesting that improper MYT1 expression affects the expression of genes involved in the cell cycle and secondary metabolite production. This study is the first to characterize a transcription factor containing a Myb DNA-binding domain that is specific to sexual development in G. zeae. PMID:21984921

  18. Starmerella meliponinorum sp. nov., a novel ascomycetous yeast species associated with stingless bees.

    PubMed

    Teixeira, Ana C P; Marini, Marjorie M; Nicoli, Jacques R; Antonini, Yasmine; Martins, Rogerio P; Lachance, Marc-André; Rosa, Carlos A

    2003-01-01

    Thirty-two strains of the novel species Starmerella meliponinorum sp. nov. were isolated from various substrates associated with three stingless bee species (tribe Meliponini) in Brazil and one in Costa Rica. The strains were found in garbage pellets (faecal material, discarded pollen or food, etc.), pollen provisions, adult bees, honey and propolis of Tetragonisca angustula, in honey from Melipona quadritasciata and in adults of Melipona rufiventris and Trigona fulviventris. The sequence of the D1/D2 domains of the large-subunit rDNA showed that the novel species belongs to the Starmerella clade and is most closely related to Candida etchellsii, although the two differ in their sequences by 7% base substitutions. S. meliponinorum is homothallic and assimilates few carbon sources. Nitrate is utilized as a sole nitrogen source. The type strain of S. meliponinorum is strain UFMG-01-J26.1T (=CBS 9117T). PMID:12656193

  19. Three New Monotypic Genera of the Caloplacoid Lichens (Teloschistaceae, Lichen-Forming Ascomycetes)

    PubMed Central

    Lőkös, Lászlo; Kim, Jung A.; Kondratiuk, Anna S.; Jeong, Min Hye; Jang, Seol Hwa; Oh, Soon-Ok; Hur, Jae-Seoun

    2015-01-01

    Three monophyletic branches are strongly supported in a phylogenetic analysis of the Teloschistaceae based on combined data sets of internal transcribed spacer and large subunit nrDNA and 12S small subunit mtDNA sequences. These are described as new monotypic genera: Jasonhuria S. Y. Kondr., L. Lőkös et S. -O. Oh, Loekoesia S. Y. Kondr., S. -O. Oh et J. -S. Hur and Olegblumia S. Y. Kondr., L. Lőkös et J. -S. Hur. Three new combinations for the type species of these genera are proposed. PMID:26539034

  20. Nutritional Composition and Bioactive Constituents of Artificial Culture of Ophiocordyceps longissima (Ascomycetes).

    PubMed

    Zhang, Ying Yu; Liu, Yang; Cheng, Wen Ming; Nam, Sung Hee; Li, Chun Ru

    2015-01-01

    In this study, the compositions of Ophiocordyceps longissima mycelia, synnemata, and fruiting bodies were first analyzed in order to clarify its chemical basis for development as a health food or medicine. We found that the contents of crude protein, polysaccharides, and macroelements were highest in mycelia, whereas effective components, including mannitol, ergosterol, adenosine, inosine, Zn, and Se, were lowest in mycelia. Polysaccharide, mannitol, and ergosterol levels in synnemata (2.33, 4.54, and 0.66 g/100 g, respectively) were similar to those in fruiting bodies, but was significantly different from those of mycelia (4.79, 1.77, and 0.43 g/100 g). Trehalose content in fruiting bodies (2.15 g/100 g) was >4 times higher than that in synnemata (0.5 g/100 g). Adenosine content in fruiting bodies (0.024 g/100 g) was 3-4 times higher than that of synnemata, whereas inosine, cytosine, guanosine, and uridine (0.093, 0.145, 0.053, and 0.073 g/100 g) were highest in synnemata. Cu and Se were lower in mycelia (3.30 10?? and 3.1 10?? g/100 g) than in synnemata (1.85 10? and 7.2 10?? g/100 g) and fruiting bodies (1.67 10? and 4.3 10?? g/100 g). As, Hg, Cd, and Pb in mycelia were under the limit of edible fungus health standard; Pb was not found, but Cd, Hg, and As were detected in fruiting bodies. These findings suggest that O. longissima could be utilized in different culture methods according to market demand and might be a possible health food or medicinal resource. PMID:25746404

  1. Intracellular siderophores are essential for ascomycete sexual development in heterothallic Cochliobolus heterostrophus and homothallic Gibberella zeae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Connections between fungal development and secondary metabolism have been reported previously, but as yet, no comprehensive analysis of a family of secondary metabolites and their possible role in fungal development has been reported. In the present study, mutant strains of the heterothallic ascomyc...

  2. Ophiostoma species (Ascomycetes: Ophiostomatales) associated with bark beetles (Coleoptera: Scolytinae) colonizing Pinus radiata in northern Spain.

    PubMed

    Romn, Pedro; Zhou, XuDong; Iturrondobeitia, Juan Carlos; Wingfield, Michael J; Goldarazena, Arturo

    2007-06-01

    Bark beetles (Coleoptera: Scolytinae) are known to be associated with fungi, especially species of Ophiostoma sensu lato and Ceratocystis. However, very little is known about these fungi in Spain. In this study, we examined the fungi associated with 13 bark beetle species and one weevil (Coleoptera: Entiminae) infesting Pinus radiata in the Basque Country of northern Spain. This study included an examination of 1323 bark beetles or their galleries in P. radiata. Isolations yielded a total of 920 cultures, which included 16 species of Ophiostoma sensu lato or their asexual states. These 16 species included 69 associations between fungi and bark beetles and weevils that have not previously been recorded. The most commonly encountered fungal associates of the bark beetles were Ophiostoma ips, Leptographium guttulatum, Ophiostoma stenoceras, and Ophiostoma piceae. In most cases, the niche of colonization had a significant effect on the abundance and composition of colonizing fungi. This confirms that resource overlap between species is reduced by partial spatial segregation. Interaction between niche and time seldom had a significant effect, which suggests that spatial colonization patterns are rarely flexible throughout timber degradation. The differences in common associates among the bark beetle species could be linked to the different niches that these beetles occupy. PMID:17668036

  3. Candida leandrae sp. nov., an asexual ascomycetous yeast species isolated from tropical plants.

    PubMed

    Ruivo, Carla C C; Lachance, Marc-André; Bacci, Maurício; Carreiro, Solange C; Rosa, Carlos A; Pagnocca, Fernando C

    2004-11-01

    The novel yeast species Candida leandrae is described based on eight isolates from decaying fruits of Leandra reversa Cogn. (Melastomataceae) in an Atlantic rainforest site in Brazil, one from a Convolvulaceae flower in Costa Rica and one from a drosophilid in Hawai'i. The strains differed in their colony morphology, one being butyrous and smooth and the other being filamentous and rugose. Sequences of the D1/D2 domains of the large-subunit rRNA gene from both morphotypes were identical. C. leandrae belongs to the Kodamaea clade and is closely related to Candida restingae. The two species can be separated on the basis of growth at 37 degrees C and the assimilation of melezitose, negative in the novel species. The type culture of C. leandrae is strain UNESP 00-64R(T) (=CBS 9735(T)=NRRL Y-27757(T)). PMID:15545490

  4. Candida ecuadorensis sp. nov., an ascomycetous yeast species found in two separate regions of Ecuador.

    PubMed

    James, Stephen A; Carvajal Barriga, Enrique Javier; Barahona, Patricia Portero; Cross, Kathryn; Bond, Christopher J; Roberts, Ian N

    2013-01-01

    In the course of an on-going study aimed at cataloguing the natural yeast biodiversity found in Ecuador, two strains (CLQCA 13-025 and CLQCA 20-004(T)) were isolated from samples of cow manure and rotten wood collected in two separate provinces of the country (Orellana and Bolvar). These strains were found to represent a novel yeast species based on the sequences of their D1/D2 domain of the large-subunit (LSU) rRNA gene and their physiological characteristics. Phylogenetic analysis based on LSU D1/D2 sequences revealed this novel species to belong to the Metschnikowia clade and to be most closely related to Candida suratensis, a species recently discovered in a mangrove forest in Thailand. The species name of Candida ecuadorensis sp. nov. is proposed to accommodate these strains, with strain CLQCA 20-004(T) (=CBS 12653(T) = NCYC 3782(T)) designated as the type strain. PMID:23104360

  5. Muscodor yucatenensis, a new endophytic ascomycete from Mexican chakah, Bursera simaruba

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During a study on the fungal endophytic associations with some trees of the dry tropical forest of El Eden Ecological Reserve located in the northeast of the Yucatan Peninsula of Mexico, a new fungal species was isolated as an endophyte of a tree named chakah, chachah or huk´up by indigenous mayas. ...

  6. Process optimization for extraction of carotenoids from medicinal caterpillar fungus, Cordyceps militaris (Ascomycetes).

    PubMed

    Yang, Tao; Sun, Junde; Lian, Tiantian; Wang, Wenzhao; Dong, Cai-Hong

    2014-01-01

    Natural carotenoids have attracted great attention for their important beneficial effects on human health and food coloring function. Cordyceps militaris, a well-known edible and medicinal fungus, is a potential source of natural carotenoids. The present study aimed to optimize the process parameters for carotenoid extraction from this mushroom. The effects of different methods of breaking the fungal cell wall and organic solvents were studied by the one-factor-at-a-time method. Subsequently, the process parameters including the duration of the extraction time, the number of extractions, and the solvent to solid ratio were optimized by using the Box-Behnken design. The optimal extraction conditions included using an acid-heating method to break the cell wall and later extracting three times, each for a 1 h duration, with a 4:1 mixture of acetone: petroleum ether and a solvent: solid ratio of 24:1. The carotenoid content varied from 2122.50 to 3847.50 µg/g dry weights in different commercially obtained fruit bodies of C. militaris. The results demonstrated that the C. militaris contained more carotenoid content in its fruit bodies than other known mushrooms. Stability monitoring by HPLC demonstrated that the carotenoids could be stored at 4°C for 40 d. It is suggested that the carotenoid content should be considered as the quality standard of commercial products of this valued mushroom. These findings will facilitate the exploration of carotenoids from C. militaris. PMID:24941034

  7. Biosynthetic investigation of phomopsins reveals a widespread pathway for ribosomal natural products in Ascomycetes.

    PubMed

    Ding, Wei; Liu, Wan-Qiu; Jia, Youli; Li, Yongzhen; van der Donk, Wilfred A; Zhang, Qi

    2016-03-29

    Production of ribosomally synthesized and posttranslationally modified peptides (RiPPs) has rarely been reported in fungi, even though organisms of this kingdom have a long history as a prolific source of natural products. Here we report an investigation of the phomopsins, antimitotic mycotoxins. We show that phomopsin is a fungal RiPP and demonstrate the widespread presence of a pathway for the biosynthesis of a family of fungal cyclic RiPPs, which we term dikaritins. We characterize PhomM as an S-adenosylmethionine-dependent α-N-methyltransferase that converts phomopsin A to anN,N-dimethylated congener (phomopsin E), and show that the methyltransferases involved in dikaritin biosynthesis have evolved differently and likely have broad substrate specificities. Genome mining studies identified eight previously unknown dikaritins in different strains, highlighting the untapped capacity of RiPP biosynthesis in fungi and setting the stage for investigating the biological activities and unknown biosynthetic transformations of this family of fungal natural products. PMID:26979951

  8. A new endophytic ascomycete from El Eden Ecological Reserve, Quintana Roo, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During a preliminary survey to report the biodiversity of endophytic fungi associated with leaves of some woody plants from El Eden Ecological Reserve in Mexico, a new fungus was isolated from Callicarpa acuminata leaves. Cultures of this fungus on PDA form a white floccose colony with a reddish-bro...

  9. The lichen-forming ascomycete Evernia mesomorpha associates with multiple genotypes of Trebouxia jamesii.

    PubMed

    Piercey-Normore, Michele D

    2006-01-01

    The epiphyte Evernia mesomorpha forms a lichen association with green algae in the genus Trebouxia. Little is known about the population structure of E. mesomorpha. Here, population structure of the algal and fungal symbionts was examined for 290 lichen thalli on 29 jack pine (Pinus banksiana) trees in Manitoba. Through phylogenetic analysis of internal transcribed spacer (ITS) nuclear ribosomal DNA (rDNA) sequences, five algal genotypes were detected that were nested within T. jamesii. Two fungal genotypes were detected that formed a clade with two other Evernia species. The genus Evernia was paraphyletic with E. prunastri, sister to Parmelia saxatilis. Restriction fragment length polymorphism (RFLP) of ITS rDNA showed multiple algal genotypes in 45% of the 290 lichen thalli collected, whereas all thalli only contained one fungal genotype. Low population subdivision of algal and fungal genotypes among trees suggested that the algal symbiont was being dispersed in the lichen soredium. Low fungal specificity for multiple algal genotypes and a hypothesized algal switch may be important life history strategies for E. mesomorpha to adapt to changing environmental conditions. PMID:16411936

  10. AbaA Regulates Conidiogenesis in the Ascomycete Fungus Fusarium graminearum

    PubMed Central

    Son, Hokyoung; Kim, Myung-Gu; Min, Kyunghun; Seo, Young-Su; Lim, Jae Yun; Choi, Gyung Ja; Kim, Jin-Cheol; Chae, Suhn-Kee; Lee, Yin-Won

    2013-01-01

    Fusarium graminearum (teleomorph Gibberella zeae) is a prominent pathogen that infects major cereal crops such as wheat, barley, and maize. Both sexual (ascospores) and asexual (conidia) spores are produced in F. graminearum. Since conidia are responsible for secondary infection in disease development, our objective of the present study was to reveal the molecular mechanisms underlying conidiogenesis in F. graminearum based on the framework previously described in Aspergillus nidulans. In this study, we firstly identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. Results from RNA-sequencing analysis suggest that AbaA plays a pivotal role in conidiation by regulating cell cycle pathways and other conidiation-related genes. Thus, the conserved roles of the AbaA ortholog in both A. nidulans and F. graminearum give new insight into the genetics of conidiation in filamentous fungi. PMID:24039821

  11. WetA Is Required for Conidiogenesis and Conidium Maturation in the Ascomycete Fungus Fusarium graminearum

    PubMed Central

    Son, Hokyoung; Kim, Myung-Gu; Min, Kyunghun; Lim, Jae Yun; Choi, Gyung Ja; Kim, Jin-Cheol; Chae, Suhn-Kee

    2014-01-01

    Fusarium graminearum, a prominent fungal pathogen that infects major cereal crops, primarily utilizes asexual spores to spread disease. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we functionally characterized the F. graminearum ortholog of Aspergillus nidulans wetA, which has been shown to be involved in conidiogenesis and conidium maturation. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidial dormancy by suppressing microcycle conidiation in F. graminearum. Transcriptome analyses demonstrated that most of the putative conidiation-related genes are expressed constitutively and that only a few genes are specifically involved in F. graminearum conidiogenesis. The conserved and distinct roles identified for WetA in F. graminearum provide new insights into the genetics of conidiation in filamentous fungi. PMID:24186953

  12. Polyphasic taxonomy of the heat resistant ascomycete genus Byssochlamys and its Paecilomyces anamorphs.

    PubMed

    Samson, R A; Houbraken, J; Varga, J; Frisvad, J C

    2009-06-01

    Byssochlamys and related Paecilomyces strains are often heat resistant and may produce mycotoxins in contaminated pasteurised foodstuffs. A comparative study of all Byssochlamys species was carried out using a polyphasic approach to find characters that differentiate species and to establish accurate data on potential mycotoxin production by each species. Phylogenetic analysis of the ITS region, parts of the beta-tubulin and calmodulin genes, macro- and micromorphological examinations and analysis of extrolite profiles were applied. Phylogenetic analyses revealed that the genus Byssochlamys includes nine species, five of which form a teleomorph, i.e. B. fulva, B. lagunculariae, B. nivea, B. spectabilis and B. zollerniae, while four are asexual, namely P. brunneolus, P. divaricatus, P. formosus and P. saturatus. Among these, B. nivea produces the mycotoxins patulin and byssochlamic acid and the immunosuppressant mycophenolic acid. Byssochlamys lagunculariae produces byssochlamic acid and mycophenolic acid and thus chemically resembles B. nivea. Some strains of P. saturatus produce patulin and brefeldin A, while B. spectabilis (anamorph P. variotii s.s.) produces viriditoxin. Some micro- and macromorphological characters are valuable for identification purposes, including the shape and size of conidia and ascospores, presence and ornamentation of chlamydospores, growth rates on MEA and CYA and acid production on CREA. A dichotomous key is provided for species identification based on phenotypical characters. PMID:20198134

  13. The Ascomycete Verticillium longisporum Is a Hybrid and a Plant Pathogen with an Expanded Host Range

    PubMed Central

    Inderbitzin, Patrik; Davis, R. Michael; Bostock, Richard M.; Subbarao, Krishna V.

    2011-01-01

    Hybridization plays a central role in plant evolution, but its overall importance in fungi is unknown. New plant pathogens are thought to arise by hybridization between formerly separated fungal species. Evolution of hybrid plant pathogens from non-pathogenic ancestors in the fungal-like protist Phytophthora has been demonstrated, but in fungi, the most important group of plant pathogens, there are few well-characterized examples of hybrids. We focused our attention on the hybrid and plant pathogen Verticillium longisporum, the causal agent of the Verticillium wilt disease in crucifer crops. In order to address questions related to the evolutionary origin of V. longisporum, we used phylogenetic analyses of seven nuclear loci and a dataset of 203 isolates of V. longisporum, V. dahliae and related species. We confirmed that V. longisporum was diploid, and originated three different times, involving four different lineages and three different parental species. All hybrids shared a common parent, species A1, that hybridized respectively with species D1, V. dahliae lineage D2 and V. dahliae lineage D3, to give rise to three different lineages of V. longisporum. Species A1 and species D1 constituted as yet unknown taxa. Verticillium longisporum likely originated recently, as each V. longisporum lineage was genetically homogenous, and comprised species A1 alleles that were identical across lineages. PMID:21455321

  14. Annulatascus aquatorba sp. nov., a lignicolous freshwater ascomycete from Sirindhorn Peat Swamp Forest, Narathiwat, Thailand.

    PubMed

    Boonyuen, Nattawut; Sri-indrasutdhi, Veera; Suetrong, Satinee; Sivichai, Somsak; Jones, E B Gareth

    2012-01-01

    As part of a long term study of fungi colonizing submerged wood in freshwater streams a new Annulatascus species, A. aquatorba, is described and illustrated from Erythrophleum teysmannii test blocks from Sirindhorn Peat Swamp Forest, southern Thailand. It differs from other species in the genus in ascospore measurements, thickness of the cell wall, 1-3-septate, fusoid to lunate shape, with central brown cells and subhyaline end cells and without a mucilaginous sheath. Asci are cylindrical, pedicellate, with a distinct, wedge-shaped and non-amyloid apical ring. Phylogenetic relationships of this species, based on the combined partial 18S and 28S rDNA, place it in the same clade as A. velatisporus (type species), A. hongkongensis and A. nilensis. PMID:22223172

  15. A 90-Day Subchronic Toxicity Study of Submerged Mycelial Culture of Cordyceps cicadae (Ascomycetes) in Rats.

    PubMed

    Chen, Yen-Lien; Yeh, Shu-Hsing; Lin, Ting-Wei; Chen, Chin-Chu; Chen, Chin-Shuh; Kuo, Chia-Feng

    2015-01-01

    Cordyceps cicadae is a parasitic fungus that hibernates inside a host (Cicada flammata Dist.) and then grows its fruiting body on the surface of the insect. The complete insect/fungus combination of C. cicadae has been widely applied in Chinese traditional medicine. Recent studies have demonstrated that the medicinal benefits of cultured mycelia are as effective as those found in the wild. However, toxicological information regarding the chronic consumption of C. cicadae mycelia culture is not available. This study was conducted to evaluate the possible toxicity arising from repeated exposure to freeze-dried submerged mycelial culture of C. cicadae for 90 days. A total of eighty 8-week-old Sprague-Dawley rats were divided into 4 groups (10 males and 10 females in each group). C. cicadae was administered daily to animals by gavage at doses of 0, 500, 1000, and 2000 mg/kg body weight for 90 days. No animal deaths occurred and no treatment-related clinical signs were observed during the study period. No statistical differences in body weight gain, relative organ weight, hematology, serum chemistry, and urinalysis were observed. Gross necropsy and histopathological findings indicated that there was no treatment-related abnormality. Based on the results, the no observed adverse effect level of C. cicadae whole broth is determined to be > 2000 mg/kg for male and female Sprague-Dawley rats. The results of this study provides support for the use of C. cicadae fermentation product as a safe agent in functional food. PMID:26559863

  16. Fruiting Body Production of the Medicinal Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes), in Artificial Medium.

    PubMed

    Cao, Li; Ye, Yunshou; Han, Richou

    2015-01-01

    Ophiocordyceps sinensis (syn. Cordyceps sinensis), regarded as the "Himalayan Viagra", is widely used for medicinal treatment and health foods. The price of O. sinensis has continued to increase over the past few years because of the growing worldwide demand and resource limitations. Artificial cultivation of the fruiting bodies to substitute natural O. sinensis is urgently needed for the effective protection of a valuable bioresource and environment in the Tibetan plateau, and for commercial trade. In this study, the anamorph of 3 isolates was separated from natural O. sinensis and identified by molecular markers as Hirsutella sinensis. These fungal isolates were cultured in a rice-based medium at 9-13°C for 50 days for mycelial growth, at 4°C for 100 days for stromatal induction, and at 13°C for 40 days for fruiting body formation. The mature fruiting bodies with mature perithecium were harvested in about 140 days. This is, to our knowledge, the first report of stable fruiting body production of O. sinensis by artificial media in the low-altitude area outside the Tibetan plateau. PMID:26853966

  17. A new endophytic ascomycete from El Eden Ecological Reserve, Quintana Roo

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During the past two years a project has been undertaken to study endophytic fungi associated with plants growing in El Eden Ecological Reserve, located in the State of Quintana Roo in the northeastern part of the Yucatan Peninsula of México. Asymptomatic, healthy leaves were collected, surface steri...

  18. Identification and characterization of polymorphic minisatellites in the phytopathogenic ascomycete Leptosphaeria maculans.

    PubMed

    Eckert, Maria; Gout, Lilian; Rouxel, Thierry; Blaise, Françoise; Jedryczka, Malgorzata; Fitt, Bruce; Balesdent, Marie-Hélène

    2005-01-01

    Leptosphaeria maculans causes phoma stem canker, the most serious disease of oilseed rape world-wide. Sexual recombination is important in the pathogen life cycle and increases the risk of plant resistance genes being overcome rapidly. Thus, there is a need to develop easy-to-use molecular markers suitable for large-scale population genetic studies. The minisatellite MinLm1, showing six alleles in natural populations, has previously been used as a marker to survey populations. Here, we report the characterization of five new minisatellites (MinLm2-MinLm6), of which four were identified by a systematic search for tandemly repeated polymorphic regions in BAC-end sequencing data from L. maculans. Of 782 BAC-end sequences analysed, 43 possessed putative minisatellite-type repeats and four of these (MinLm3-MinLm6) displayed both consistent PCR amplification and size polymorphism in a collection of L. maculans isolates of diverse origins. Cloning and sequencing of each allele confirmed that polymorphism was due to variation in the repeat number of a core motif ranging from 11 bp (MinLm3) to 51 bp (MinLm4). The number of alleles found for each minisatellite ranged from three (MinLm4) to nine (MinLm2), with eight, five and six for MinLm3, MinLm5 and MinLm6, respectively. MinLm2-MinLm6 are all single locus markers specific to L. maculans and share some common features, such as conservation of core motifs and incomplete direct repeats in the flanking regions. To our knowledge, L. maculans is the first fungal species for which six polymorphic single locus minisatellite markers have been reported. PMID:15614492

  19. Mesosynteny; A novel mode of chromosomal evolution peculiar to filamentous Ascomycete fungi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report a novel form of evolution in which genes are conserved within homologous chromosomes, but with randomised orders and orientations. We propose to call this mode of evolution 'mesosynteny'. Mesosynteny is an alternative evolutionary pathway to macrosyntenic conservation. Mesosynteny would ...

  20. Remispora spitsbergenensis sp. nov., a marine lignicolous ascomycete from Svalbard, Norway.

    PubMed

    Pang, Ka-Lai; Chiang, Michael W L; Vrijmoed, Lilian L P

    2009-01-01

    Abstract: Remispora was established for R. maritima, a fungus with globose/subglobose, lightly colored and coriaceous ascomata; deliquescing asci; ellipsoidal ascospores; and bipolar, pleomorphic ascospore appendages. Seven species currently are included in Remispora: R crispa, R. galerita, R maritima, R. minuta, R. pilleata, R. quadriremis and R stellata. Variations on ascospore appendages can be observed in Remispora. In general the appendage is exosporic in nature and comprises an amorphous, electron-transparent matrix, and a fibrous, electron-dense component. An eighth Remispora species, R. spitsbergenensis sp. nov., is described here, discovered from washed-up wood collected at the shore of Longyearbyen, Svalbard, Norway. Ascospore appendages of R. spitsbergenensis appear as fibrous strands and amorphic material under the scanning electron microscope, which are characteristic of a Remispora species. Remispora spitsbergenensis resembles R. quadriremis and R. stellata because all possess four or more ascospore appendages at one end. Remispora spitsbergenensis possesses consistently four polar appendages at each end in contrast to six in R. stellata. Also ascospore appendages of R. spitsbergenensis are ribbon-like, compared with the obclavate, curved and attenuate appendages in R. quadriremis and R. stellata. A key for the identification of the eight Remispora species is provided. PMID:19623932

  1. New Species and New Records of Buellia (Lichenized Ascomycetes) from Jeju Province, South Korea.

    PubMed

    Wang, Xin Yu; Liu, Dong; Lőkös, László; Kondratyuk, Sergey Y; Oh, Soon-Ok; Park, Jung Shin; Hur, Jae-Seoun

    2016-03-01

    A new species and 2 new records of lichen genus Buellia were discovered from Chuja-do Island in Jeju Province during a recent floristic survey: B. chujana X. Y. Wang, S. Y. Kondr., L. Lőkös & J.-S. Hur sp. nov., B. halonia (Ach.) Tuck., and B. mamillana (Tuck.) W. A. Weber. The new species is characterized by a brown, areolate thallus, the presence of perlatolic acid, and a saxicolous habitat. Together with previously recorded species, 10 Buellia species were confirmed from Jeju-do Island. Among these species, 3 growing in the exposed rocky area contained xanthone (yellowish lichen thallus, UV + orange), indicating that production of xanthone in this genus might be a defense strategy against the harm of UV light. Although the genus Buellia has been thoroughly studied in Korea before, novel species have been discovered continuously, and large species diversity has been found in this crustose genus, even from a small rocky island. This study indicates that the coastal area harbors a vast number of crustose lichen species, and there is great potential to discover unknown lichens in the coastal rocky area in Korea. PMID:27103850

  2. Evaluation of automated cell disruptor methods for oomycetous and ascomycetous model organisms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two automated cell disruptor-based methods for RNA extraction; disruption of thawed cells submerged in TRIzol Reagent (method QP), and direct disruption of frozen cells on dry ice (method CP), were optimized for a model oomycete, Phytophthora capsici, and compared with grinding in a mortar and pestl...

  3. New Species and New Records of Buellia (Lichenized Ascomycetes) from Jeju Province, South Korea

    PubMed Central

    Wang, Xin Yu; Liu, Dong; Lőkös, László; Kondratyuk, Sergey Y.; Oh, Soon-Ok; Park, Jung Shin

    2016-01-01

    A new species and 2 new records of lichen genus Buellia were discovered from Chuja-do Island in Jeju Province during a recent floristic survey: B. chujana X. Y. Wang, S. Y. Kondr., L. Lőkös & J.-S. Hur sp. nov., B. halonia (Ach.) Tuck., and B. mamillana (Tuck.) W. A. Weber. The new species is characterized by a brown, areolate thallus, the presence of perlatolic acid, and a saxicolous habitat. Together with previously recorded species, 10 Buellia species were confirmed from Jeju-do Island. Among these species, 3 growing in the exposed rocky area contained xanthone (yellowish lichen thallus, UV + orange), indicating that production of xanthone in this genus might be a defense strategy against the harm of UV light. Although the genus Buellia has been thoroughly studied in Korea before, novel species have been discovered continuously, and large species diversity has been found in this crustose genus, even from a small rocky island. This study indicates that the coastal area harbors a vast number of crustose lichen species, and there is great potential to discover unknown lichens in the coastal rocky area in Korea. PMID:27103850

  4. Muscodor yucatanensis, a new endophytic ascomycete from Mexican chakah, Bursera simaruba

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During a study on the fungal endophytic associations with some trees of the secondary forest of El Eden Ecological Reserve located in the northeastern Yucatan Peninsula of Mexico, a new fungal species was isolated as an endophyte of a tree named chakah, chachah or hukúp (Bursera simaruba) by indigen...

  5. Transposable group II introns in fission and budding yeast. Site-specific genomic instabilities and formation of group II IVS plDNAs.

    PubMed

    Schmidt, W M; Schweyen, R J; Wolf, K; Mueller, M W

    1994-10-21

    The recent report on RNA-mediated group II intron (IVS, intervening sequence) transposition in mitochondria (mt) of Saccharomyces cerevisiae and Podospora anserina and the demonstration of reverse transcriptase (RT) activity encoded by the mobile S. cerevisiae intron cox1-aI1 suggests that group II introns constitute a new class of site-specific retro-like (retroid) elements. This is supported by the finding that the mitochondrial cob1-bI1 intron from the fission yeast Schizosaccharomyces pombe, encoding an RT-like open reading frame, is transposed in mtDNA populations. In agreement with the involvement of an RNA-intermediate in IVS transposition: First, the insertion sites were preceded by at least an IBS1-like (intron binding site) motif, which corresponds to the upstream exon and suffices to form the IBS1/EBS1 (EBS: exon binding site) base-pairing interactions. Second, intron transposition was conservative with respect to sequences flanking the insertion sites. We formulated the hypothesis that transient IVS insertion at non-allelic sites followed by recombination can be viewed as a general molecular mechanism, applicable equally well to site-specific genomic instabilities involving splice-site borders of group II introns and to the formation of extra-genomic IVS plasmid DNAs (plDNAs). We used polymerase chain reaction (PCR) techniques to detect infrequent rearrangements in mtDNA and report here on duplicative IVS transposition, twintron formation (e.g. bI1 insertion into another bI1 intron), and IVS insertions at canonical 5' exon-intron borders in S. pombe (cob1-bI1) and in S. cerevisiae (cox1-aI1). These data substantiate the concept that group II intron homing, IVS transposition and circular IVS plDNA formation involve a common RNA-mediated mechanism. Finally, the findings suggest that extra-genomic group II IVS copies are not restricted to senescence mycelia of P. anserina, but constitute natural components of group II IVS-containing genomes. PMID:7932746

  6. Signal Transduction by a Fungal NOD-Like Receptor Based on Propagation of a Prion Amyloid Fold

    PubMed Central

    Daskalov, Asen; Habenstein, Birgit; Martinez, Denis; Debets, Alfons J. M.; Sabaté, Raimon; Loquet, Antoine; Saupe, Sven J.

    2015-01-01

    In the fungus Podospora anserina, the [Het-s] prion induces programmed cell death by activating the HET-S pore-forming protein. The HET-s β-solenoid prion fold serves as a template for converting the HET-S prion-forming domain into the same fold. This conversion, in turn, activates the HET-S pore-forming domain. The gene immediately adjacent to het-S encodes NWD2, a Nod-like receptor (NLR) with an N-terminal motif similar to the elementary repeat unit of the β-solenoid fold. NLRs are immune receptors controlling cell death and host defense processes in animals, plants and fungi. We have proposed that, analogously to [Het-s], NWD2 can activate the HET-S pore-forming protein by converting its prion-forming region into the β-solenoid fold. Here, we analyze the ability of NWD2 to induce formation of the β-solenoid prion fold. We show that artificial NWD2 variants induce formation of the [Het-s] prion, specifically in presence of their cognate ligands. The N-terminal motif is responsible for this prion induction, and mutations predicted to affect the β-solenoid fold abolish templating activity. In vitro, the N-terminal motif assembles into infectious prion amyloids that display a structure resembling the β-solenoid fold. In vivo, the assembled form of the NWD2 N-terminal region activates the HET-S pore-forming protein. This study documenting the role of the β-solenoid fold in fungal NLR function further highlights the general importance of amyloid and prion-like signaling in immunity-related cell fate pathways. PMID:25671553

  7. High natural prevalence of a fungal prion

    PubMed Central

    Debets, Alfons J. M.; Dalstra, Henk J. P.; Slakhorst, Marijke; Koopmanschap, Bertha; Hoekstra, Rolf F.; Saupe, Sven J.

    2012-01-01

    Prions are infectious proteins that cause fatal diseases in mammals. Prions have also been found in fungi, but studies on their role in nature are scarce. The proposed biological function of fungal prions is debated and varies from detrimental to benign or even beneficial. [Het-s] is a prion of the fungus Podospora anserina. The het-s locus exists as two antagonistic alleles that constitute an allorecognition system: the het-s allele encoding the protein variant capable of prion formation and the het-S allele encoding a protein variant that cannot form a prion. We document here that het-s alleles, capable of prion formation, are nearly twice as frequent as het-S alleles in a natural population of 112 individuals. Then, we report a 92% prevalence of [Het-s] prion infection among the het-s isolates and find evidence of the role of the [Het-s]/het-S allorecognition system on the incidence of infection by a deleterious senescence plasmid. We explain the het-s/het-S allele ratios by the existence of two selective forces operating at different levels. We propose that during the somatic stage, the role of [Het-s]/HET-S in allorecognition leads to frequency-dependent selection for which an equilibrated frequency would be optimal. However, in the sexual cycle, the [Het-s] prion causes meiotic drive favoring the het-s allele. Our findings indicate that [Het-s] is a selected and, therefore, widespread prion whose activity as selfish genetic element is counteracted by balancing selection for allorecognition polymorphism. PMID:22691498

  8. Conformational folding and stability of HET-C2 glycolipid transfer protein (GLTP)-fold: Does a molten globule-like state regulate activity?†

    PubMed Central

    Kenoth, Roopa; Kamlekar, Ravi Kanth; Simanshu, Dhirendra K.; Gao, Yongguang; Malinina, Lucy; Prendergast, Franklyn G.; Molotkovsky, Julian G.; Patel, Dinshaw J.; Venyaminov, Sergei Y.; Brown, Rhoderick E.

    2011-01-01

    The glycolipid transfer protein (GLTP) superfamily is defined by the human GLTP-fold which represents a novel motif for lipid binding/transfer and for reversible interaction with membranes, i.e. peripheral amphitropic proteins. Despite limited sequence homology with human GLTP, we recently showed that HET-C2 GLTP of Podospora anserina is organized conformationally as a GLTP-fold. Currently, insights into the folding stability and conformational states that regulate GLTP-fold activity are almost nonexistent. To gain such insights into the disulfide-less GLTP-fold, we investigated the effect of changing pH on the fungal HET-C2 GLTP-fold by taking advantage of its two tryptophans and four tyrosines (compared to three tryptophans and ten tyrosines in human GLTP). pH-Induced conformational alterations were determined by changes in: i) intrinsic tryptophan fluorescence (intensity, emission wavelength maximum, anisotropy); ii) circular dichroism over the near-UV and far-UV ranges including thermal stability profiles of the derivatized molar ellipticity at 222 nm; iii) fluorescence properties of 1-anilinonaphthalene-8-sulfonic acid; iv) glycolipid inter-membrane transfer activity monitored by Förster resonance energy transfer. Analyses of our recently solved crystallographic structure of HET-C2 (1.9 Å) enabled identification of side chain electrostatic interactions that contribute to HET-C2 GLTP-fold stability and can be altered by pH change. Side-chain interactions include numerous salt bridges and interchain cation-pi interactions, but not intramolecular disulfide bridges. Histidine residues are especially important for stabilizing the local positioning of the two tryptophan residues and the conformation of adjacent chains. Induction of a low pH-induced, molten globule-like state inhibited glycolipid intermembrane transfer by the HET-C2 GLTP-fold. PMID:21553912

  9. STITCHER: Dynamic assembly of likely amyloid and prion β-structures from secondary structure predictions

    PubMed Central

    Bryan, Allen W; O’Donnell, Charles W; Menke, Matthew; Cowen, Lenore J; Lindquist, Susan; Berger, Bonnie

    2012-01-01

    The supersecondary structure of amyloids and prions, proteins of intense clinical and biological interest, are difficult to determine by standard experimental or computational means. In addition, significant conformational heterogeneity is known or suspected to exist in many amyloid fibrils. Previous work has demonstrated that probability-based prediction of discrete β-strand pairs can offer insight into these structures. Here, we devise a system of energetic rules that can be used to dynamically assemble these discrete β-strand pairs into complete amyloid β-structures. The STITCHER algorithm progressively ‘stitches’ strand-pairs into full β-sheets based on a novel free-energy model, incorporating experimentally observed amino-acid side-chain stacking contributions, entropic estimates, and steric restrictions for amyloidal parallel β-sheet construction. A dynamic program computes the top 50 structures and returns both the highest scoring structure and a consensus structure taken by polling this list for common discrete elements. Putative structural heterogeneity can be inferred from sequence regions that compose poorly. Predictions show agreement with experimental models of Alzheimer’s amyloid beta peptide and the Podospora anserina Het-s prion. Predictions of the HET-s homolog HET-S also reflect experimental observations of poor amyloid formation. We put forward predicted structures for the yeast prion Sup35, suggesting N-terminal structural stability enabled by tyrosine ladders, and C-terminal heterogeneity. Predictions for the Rnq1 prion and alpha-synuclein are also given, identifying a similar mix of homogenous and heterogeneous secondary structure elements. STITCHER provides novel insight into the energetic basis of amyloid structure, provides accurate structure predictions, and can help guide future experimental studies. Proteins 2012. © 2011 Wiley Periodicals, Inc. PMID:22095906

  10. Albumin impairs renal tubular tight junctions via targeting the NLRP3 inflammasome.

    PubMed

    Zhuang, Yibo; Hu, Caiyu; Ding, Guixia; Zhang, Yue; Huang, Songming; Jia, Zhanjun; Zhang, Aihua

    2015-05-01

    Proteinuria is, not only a hallmark of glomerular disease, but also a contributor to kidney injury. However, its pathogenic mechanism is still elusive. In the present study, the effects of albumin on renal tubular tight junctions and the potential molecular mechanisms of those effects were investigated. In mouse proximal tubular cells (mPTCs), albumin treatment resulted in a significant loss of the cellular tight junction proteins zonula occludens-1 (ZO-1) and claudin-1 in a time- and dose-dependent manner, indicating a severe impairment of the tight junctions. On the basis of our previous study showing that albumin stimulated NLRP3 [neuronal apoptosis inhibitor protein, major histocompatibility complex class 2 transcription activator, incompatibility locus protein from Podospora anserina, and telomerase-associated protein (NACHT); leucine-rich repeat (LRR); and pyrin domain (PYD) domains-containing protein 3] inflammasome activation in mPTCs, we pretreated mPTCs with NLRP3 siRNA (siNLRP3) and found that NLRP3 knockdown significantly blocked the downregulation of ZO-1 and claudin-1 induced by albumin. Similarly, in albumin-overloaded wild-type mice, both ZO-1 and claudin-1 were downregulated at the protein and mRNA levels in parallel with the impaired formation of the tight junctions on transmission electron microscopy and the abnormal renal tubular morphology on periodic acid-Schiff staining, which contrasted with the stimulation of NLRP3 in the renal tubules. In contrast, NLRP3 knockout (NLRP3(-/-)) mice preserved normal ZO-1 and claudin-1 expression as well as largely normal tight junctions and tubular morphology. More importantly, deletion of the NLRP3 pathway downstream component caspase-1 similarly blocked the albumin overload-induced downregulation of ZO-1 and claudin-1. Taken together, these findings demonstrated an important role of the albumin-NLRP3 inflammasome axis in mediating the impairment of renal tubular tight junctions and integrity. PMID:25715986

  11. Distribution and evolution of het gene homologs in the basidiomycota.

    PubMed

    Van der Nest, M A; Olson, A; Lind, M; Vélëz, H; Dalman, K; Brandström Durling, M; Karlsson, M; Stenlid, J

    2014-03-01

    In filamentous fungi a system known as somatic incompatibility (SI) governs self/non-self recognition. SI is controlled by a regulatory signaling network involving proteins encoded at the het (heterokaryon incompatible) loci. Despite the wide occurrence of SI, the molecular identity and structure of only a small number of het genes and their products have been characterized in the model fungi Neurospora crassa and Podospora anserina. Our aim was to identify and study the distribution and evolution of putative het gene homologs in the Basidiomycota. For this purpose we used the information available for the model fungi to identify homologs of het genes in other fungi, especially the Basidiomycota. Putative het-c, het-c2 and un-24 homologs, as well as sequences containing the NACHT, HET or WD40 domains present in the het-e, het-r, het-6 and het-d genes were identified in certain members of the Ascomycota and Basidiomycota. The widespread phylogenetic distribution of certain het genes may reflect the fact that the encoded proteins are involved in fundamental cellular processes other than SI. Although homologs of het-S were previously known only from the Sordariomycetes (Ascomycota), we also identified a putative homolog of this gene in Gymnopus luxurians (Basidiomycota, class Agaricomycetes). Furthermore, with the exception of un-24, all of the putative het genes identified occurred mostly in a multi-copy fashion, some with lineage and species-specific expansions. Overall our results indicated that gene duplication followed by gene loss and/or gene family expansion, as well as multiple events of domain fusion and shuffling played an important role in the evolution of het gene homologs of Basidiomycota and other filamentous fungi. PMID:24380733

  12. STITCHER: Dynamic assembly of likely amyloid and prion β-structures from secondary structure predictions.

    PubMed

    Bryan, Allen W; O'Donnell, Charles W; Menke, Matthew; Cowen, Lenore J; Lindquist, Susan; Berger, Bonnie

    2012-02-01

    The supersecondary structure of amyloids and prions, proteins of intense clinical and biological interest, are difficult to determine by standard experimental or computational means. In addition, significant conformational heterogeneity is known or suspected to exist in many amyloid fibrils. Previous work has demonstrated that probability-based prediction of discrete β-strand pairs can offer insight into these structures. Here, we devise a system of energetic rules that can be used to dynamically assemble these discrete β-strand pairs into complete amyloid β-structures. The STITCHER algorithm progressively 'stitches' strand-pairs into full β-sheets based on a novel free-energy model, incorporating experimentally observed amino-acid side-chain stacking contributions, entropic estimates, and steric restrictions for amyloidal parallel β-sheet construction. A dynamic program computes the top 50 structures and returns both the highest scoring structure and a consensus structure taken by polling this list for common discrete elements. Putative structural heterogeneity can be inferred from sequence regions that compose poorly. Predictions show agreement with experimental models of Alzheimer's amyloid beta peptide and the Podospora anserina Het-s prion. Predictions of the HET-s homolog HET-S also reflect experimental observations of poor amyloid formation. We put forward predicted structures for the yeast prion Sup35, suggesting N-terminal structural stability enabled by tyrosine ladders, and C-terminal heterogeneity. Predictions for the Rnq1 prion and alpha-synuclein are also given, identifying a similar mix of homogenous and heterogeneous secondary structure elements. STITCHER provides novel insight into the energetic basis of amyloid structure, provides accurate structure predictions, and can help guide future experimental studies. PMID:22095906

  13. High natural prevalence of a fungal prion.

    PubMed

    Debets, Alfons J M; Dalstra, Henk J P; Slakhorst, Marijke; Koopmanschap, Bertha; Hoekstra, Rolf F; Saupe, Sven J

    2012-06-26

    Prions are infectious proteins that cause fatal diseases in mammals. Prions have also been found in fungi, but studies on their role in nature are scarce. The proposed biological function of fungal prions is debated and varies from detrimental to benign or even beneficial. [Het-s] is a prion of the fungus Podospora anserina. The het-s locus exists as two antagonistic alleles that constitute an allorecognition system: the het-s allele encoding the protein variant capable of prion formation and the het-S allele encoding a protein variant that cannot form a prion. We document here that het-s alleles, capable of prion formation, are nearly twice as frequent as het-S alleles in a natural population of 112 individuals. Then, we report a 92% prevalence of [Het-s] prion infection among the het-s isolates and find evidence of the role of the [Het-s]/het-S allorecognition system on the incidence of infection by a deleterious senescence plasmid. We explain the het-s/het-S allele ratios by the existence of two selective forces operating at different levels. We propose that during the somatic stage, the role of [Het-s]/HET-S in allorecognition leads to frequency-dependent selection for which an equilibrated frequency would be optimal. However, in the sexual cycle, the [Het-s] prion causes meiotic drive favoring the het-s allele. Our findings indicate that [Het-s] is a selected and, therefore, widespread prion whose activity as selfish genetic element is counteracted by balancing selection for allorecognition polymorphism. PMID:22691498

  14. Metabolic engineering of muconic acid production in Saccharomyces cerevisiae.

    PubMed

    Curran, Kathleen A; Leavitt, John M; Karim, Ashty S; Alper, Hal S

    2013-01-01

    The dicarboxylic acid muconic acid has garnered significant interest due to its potential use as a platform chemical for the production of several valuable consumer bio-plastics including nylon-6,6 and polyurethane (via an adipic acid intermediate) and polyethylene terephthalate (PET) (via a terephthalic acid intermediate). Many process advantages (including lower pH levels) support the production of this molecule in yeast. Here, we present the first heterologous production of muconic acid in the yeast Saccharomyces cerevisiae. A three-step synthetic, composite pathway comprised of the enzymes dehydroshikimate dehydratase from Podospora anserina, protocatechuic acid decarboxylase from Enterobacter cloacae, and catechol 1,2-dioxygenase from Candida albicans was imported into yeast. Further genetic modifications guided by metabolic modeling and feedback inhibition mitigation were introduced to increase precursor availability. Specifically, the knockout of ARO3 and overexpression of a feedback-resistant mutant of aro4 reduced feedback inhibition in the shikimate pathway, and the zwf1 deletion and over-expression of TKL1 increased flux of necessary precursors into the pathway. Further balancing of the heterologous enzyme levels led to a final titer of nearly 141mg/L muconic acid in a shake-flask culture, a value nearly 24-fold higher than the initial strain. Moreover, this strain has the highest titer and second highest yield of any reported shikimate and aromatic amino acid-based molecule in yeast in a simple batch condition. This work collectively demonstrates that yeast has the potential to be a platform for the bioproduction of muconic acid and suggests an area that is ripe for future metabolic engineering efforts. PMID:23164574

  15. Efficacy of entomopathogenic fungi (Ascomycetes: Hypocreales) against adult Haematobia irritans (Diptera: Muscidae) under stable conditions in the Mexican dry tropics.

    PubMed

    Galindo-Velasco, E; Lezama-Gutiérrez, R; Cruz-Vázquez, C; Pescador-Rubio, A; Angel-Sahagún, C A; Ojeda-Chi, M M; Rodríguez-Vivas, R I; Contreras-Lara, D

    2015-04-30

    The purpose of this paper is to evaluate the effect of five strains of Metarhizium anisopliae (Ma) and three strains of Isaria fumosorosea (Ifr) at a concentration of 1×10(8)colony-forming units/ml applied by spraying onto bovines with controlled infestation of Haematobia irritans under stable conditions in the Mexican dry tropics. Four experiments were performed, in each of which three treatments (two fungal strains and one control) were evaluated with eight repetitions for each one, by carrying out a single application of the aqueous suspension of each strain. The animals were isolated in individual cages and direct counts of the infestation were carried out for 13 days. It was observed that strains Ma2, Ma6, Ma10, Ma14, and Ma34 caused 94-100% reduction in infestation between days 12 and 13 post-treatment, while strains Ifr19, Ifr11, and Ifr12 reduced infestation from 90% to 98% up to day 13 post-application. There was an effect in the generation of horn flies from the excrement of bovines that were treated with different strains, reducing the reproduction of subsequent generations. It was concluded that the strains of M. anisopliae and I. fumosorosea evaluated in this study can be used as biocontrol agents in infestations of H. irritans in stabled bovines. PMID:25771932

  16. A transcriptomic study of grapevine (Vitis vinifera cv. Cabernet-Sauvignon) interaction with the vascular ascomycete fungus Eutypa lata

    PubMed Central

    Camps, Céline; Kappel, Christian; Lecomte, Pascal; Léon, Céline; Gomès, Eric; Coutos-Thévenot, Pierre; Delrot, Serge

    2010-01-01

    Eutypa dieback is a vascular disease that may severely affect vineyards throughout the world. In the present work, microarrays were made in order (i) to improve our knowledge of grapevine (Vitis vinifera cv. Cabernet-Sauvignon) responses to Eutypa lata, the causal agent of Eutypa dieback; and (ii) to identify genes that may prevent symptom development. Qiagen/Operon grapevine microarrays comprising 14 500 probes were used to compare, under three experimental conditions (in vitro, in the greenhouse, and in the vineyard), foliar material of infected symptomatic plants (S+R+), infected asymptomatic plants (S–R+), and healthy plants (S–R–). These plants were characterized by symptom notation after natural (vineyard) or experimental (in vitro and greenhouse) infection, re-isolation of the fungus located in the lignified parts, and the formal identification of E. lata mycelium by PCR. Semi-quantitative real-time PCR experiments were run to confirm the expression of some genes of interest in response to E. lata. Their expression profiles were also studied in response to other grapevine pathogens (Erysiphe necator, Plasmopara viticola, and Botrytis cinerea). (i) Five functional categories of genes, that is those involved in metabolism, defence reactions, interaction with the environment, transport, and transcription, were up-regulated in S+R+ plants compared with S–R– plants. These genes, which cannot prevent infection and symptom development, are not specific since they were also up-regulated after infection by powdery mildew, downy mildew, and black rot. (ii) Most of the genes that may prevent symptom development are associated with the light phase of photosynthesis. This finding is discussed in the context of previous data on the mode of action of eutypin and the polypeptide fraction secreted by Eutypa. PMID:20190040

  17. Genome and physiology of the ascomycete filamentous fungus Xeromyces bisporus, the most xerophilic organism isolated to date.

    PubMed

    Leong, Su-Lin L; Lantz, Henrik; Pettersson, Olga V; Frisvad, Jens C; Thrane, Ulf; Heipieper, Hermann J; Dijksterhuis, Jan; Grabherr, Manfred; Pettersson, Mats; Tellgren-Roth, Christian; Schnrer, Johan

    2015-02-01

    Xeromyces bisporus can grow on sugary substrates down to 0.61, an extremely low water activity. Its genome size is approximately 22?Mb. Gene clusters encoding for secondary metabolites were conspicuously absent; secondary metabolites were not detected experimentally. Thus, in its 'dry' but nutrient-rich environment, X.?bisporus appears to have relinquished abilities for combative interactions. Elements to sense/signal osmotic stress, e.g. HogA pathway, were present in X.?bisporus. However, transcriptomes at optimal (??0.89) versus low aw (0.68) revealed differential expression of only a few stress-related genes; among these, certain (not all) steps for glycerol synthesis were upregulated. Xeromyces bisporus increased glycerol production during hypo- and hyper-osmotic stress, and much of its wet weight comprised water and rinsable solutes; leaked solutes may form a protective slime. Xeromyces bisporus and other food-borne moulds increased membrane fatty acid saturation as water activity decreased. Such modifications did not appear to be transcriptionally regulated in X.?bisporus; however, genes modulating sterols, phospholipids and the cell wall were differentially expressed. Xeromyces bisporus was previously proposed to be a 'chaophile', preferring solutes that disorder biomolecular structures. Both X.?bisporus and the closely related xerophile, Xerochrysium xerophilum, with low membrane unsaturation indices, could represent a phylogenetic cluster of 'chaophiles'. PMID:25142400

  18. Cell polarity and hyphal morphogenesis are controlled by multiple rho-protein modules in the filamentous ascomycete Ashbya gossypii.

    PubMed Central

    Wendland, J; Philippsen, P

    2001-01-01

    Polarized cell growth requires a polarized organization of the actin cytoskeleton. Small GTP-binding proteins of the Rho-family have been shown to be involved in the regulation of actin polarization as well as other processes. Hyphal growth in filamentous fungi represents an ideal model to investigate mechanisms involved in generating cell polarity and establishing polarized cell growth. Since a potential role of Rho-proteins has not been studied so far in filamentous fungi we isolated and characterized the Ashbya gossypii homologs of the Saccharomyces cerevisiae CDC42, CDC24, RHO1, and RHO3 genes. The AgCDC42 and AgCDC24 genes can both complement conditional mutations in the S. cerevisiae CDC42 and CDC24 genes and both proteins are required for the establishment of actin polarization in A. gossypii germ cells. Agrho1 mutants show a cell lysis phenotype. Null mutant strains of Agrho3 show periodic swelling of hyphal tips that is overcome by repolarization and polar hyphal growth in a manner resembling the germination pattern of spores. Thus different Rho-protein modules are required for distinct steps during polarized hyphal growth of A. gossypii. PMID:11156982

  19. Antioxidant and antiradical properties of methanolic extracts from algerian wild edible desert truffles (terfezia and tirmania, ascomycetes).

    PubMed

    Gouzi, Hicham; Leboukh, Mourad; Bouchouka, Elmouloud

    2013-01-01

    Wild edible truffles (namely, Terfezia leonis, Tirmania pinoyi, and T. nivea) were commercially obtained from Southern Algeria and methanolic extracts were prepared from these truffles. Their antioxidant and antiradical properties were studied by using five analytical methods: scavenging capacity on 1,1-diphenyl-2-picrylhydrazyl (DPPH·), 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS·+), superoxide anion (O2·-) radicals, ferric reducing antioxidant power (FRAP), and ferricyanide/Prussian blue assay. Chemical constituents contributing to these activities were also investigated. T. leonis had the highest total phenolics, total carotenoids, and anthocyanin contents. At 2.6 mg/mL, scavenging effects on the DPPH· radical were 92.47%, 53.06%, and 41.34% for T. leonis, T. pinoyi, and T. nivea, respectively. T. leonis showed the most potent radical scavenging activities on DPPH·, ABTS·+, and O2·- radicals, with EC50 values of 1.08, 1.35, and 7.27 mg/mL, respectively. On the other hand, T. leonis exhibited the highest reductive capabilities. On the basis of the EC50 values, T. leonis had good antioxidant and antiradical properties. These results showed that methanolic extracts from these three truffles species had effective antioxidant and antiradical properties. Therefore, wild edible desert truffles could serve as an easily accessible item of food rich in natural antioxidants, as a possible food supplement, or even as a pharmaceutical agent. PMID:24266372

  20. PHYLOGENETIC ANALYSIS IDENTIFIES THE "MEGABACTERIUM" OF BIRDS AS A NOVEL ANAMORPHIC ASCOMYCETOUS YEAST, MACRORHABDUS ORNITHOGASTER GEN. NOV, SP. NOV.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Organisms commonly referred to as megabacteria colonize the gastric isthmus of many species of birds. They are weakly Gram-positive, PAS-positive, and stain with silver stains. Previous studies show that they have a nucleus and a cell wall similar to that seen in fungi. Calcofluor white M2R stain...

  1. On the reclassification of species assigned to Candida and other anamorphic ascomycetous yeast genera based on phylogenetic circumscription

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multigene phylogenies have been instrumental in revising the classification of ascosporic (teleomorph) yeasts in a natural system based on lines of decent. Although many taxonomic changes have already been implemented for teleomorph taxa, this is not yet the case for the large genus Candida and smal...

  2. Kazachstania yasuniensis sp. nov., an ascomycetous yeast species found in mainland Ecuador and on the Galápagos.

    PubMed

    James, Stephen A; Carvajal Barriga, Enrique Javier; Portero Barahona, Patricia; Nueno-Palop, Carmen; Cross, Kathryn; Bond, Christopher J; Roberts, Ian N

    2015-04-01

    Seven strains representing a novel yeast species belonging to the genus Kazachstania were found at several collection sites on both mainland Ecuador (Yasuní National Park) and the Galápagos (Santa Cruz Island). Two strains (CLQCA 20-132(T) and CLQCA 24SC-045) were isolated from rotten wood samples, two further strains (CLQCA 20-280 and CLQCA 20-348) were isolated from soil samples, and three strains (CLQCA 20-198, CLQCA 20-374 and CLQCA 20-431) were isolated from decaying fruits. Sequence analyses of the D1/D2 domains of the LSU rRNA gene and ribosomal internal transcribed spacer (ITS) region indicated that the novel species is most closely related to Kazachstania servazzii and Kazachstania unispora. Although the strains could not be distinguished from one another based upon their differing geographical origins, they could be differentiated according to their isolation source (fruit, soil or wood) by ITS sequencing. The species name Kazachstania yasuniensis sp. nov. is proposed to accommodate these strains, with CLQCA 20-132(T) ( = CBS 13946(T) = NCYC 4008(T)) designated the type strain. PMID:25644482

  3. Cordyceps s.l. (Ascomycetes) Species Used as Medicinal Mushrooms Are Closely Related with Higher Ability to Produce Cordycepin.

    PubMed

    Kuo, Hsiao-Che; Huang, I-Ching; Chen, Tzong-Yueh

    2015-01-01

    Cordyceps s.l. (sensu lato) species have been used as herbal medicines; one of their main constituents is cordycepin. As genome sequencing techniques have become more cost-effective and more popular, more entomogenous fungal genomes have been sequenced and published. Here, we constructed a phylogenetic tree based on 18S rRNA sequences from Cordyceps species and analyzed the copy number of the key enzymes involved in biosynthesis of cordycepin from related fungal genomes that have been published. The sequences of the 18S rRNA gene were examined, and seven single nucleotides were found that could represent the evolutionary history of Cordyceps s.l. and which perfectly fit the phylogenetic tree. Their evolution was influenced mainly by host factors, rather than geographical location. The Cordyceps s.l. species that are used as herbal medicines are closely related in the phylogenetic tree. The major species for Chinese pharmaceutical markets, such as C. militaris and C. sinensis, have higher copy numbers of 5'-nucleotidase and adenylate kinase, and ribonucleotide reductases (RNRs), respectively. Moreover, absence of an RNR inhibitor may cause cordycepin accumulation. Presence of an RNR inhibitor may lead to lower cordycepin levels in fungal species in which no medicinal applications have been described. Cordycepin is not only an important secondary metabolite that is used as an herbal medicine, but it also has significance for understanding the evolution of these entomogenous species. PMID:26853963

  4. Review on Natural Enemies and Diseases in the Artificial Cultivation of Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes).

    PubMed

    Lu, Zenghui; Shi, Ping; He, Yuanchuan; Zhang, Deli; He, Zongyi; Chen, Shijiang; Tu, Yongqin; Li, Li; Liu, Fei; Zeng, Wei

    2015-01-01

    Ophiocordyceps sinensis (syn. Cordyceps sinensis), well known as DongChongXiaCao (DCXC), is one of the most valuable traditional Chinese medicinal species. In this article, we provide a systematic review of natural enemies and diseases encountered in artificial cultivation of DCXC. Unfortunately, DCXC has been endangered over the past decades due to overharvesting and a worsening ecological environment. Therefore, the artificial cultivation of DCXC has been extensively investigated in recent years. Complete indoor artificial cultivation and semi-field cultivation are the two most common strategies used to cultivate DCXC. However, cultured DCXCs are often attacked by various natural enemies and diseases, which have resulted in substantial loss of the valuable medicinal resource. In this study, we have summarized the species of natural enemies and types of diseases confronted by DCXC. Twenty reported natural enemy species are categorized into four classes, one of which is reported for the first time in this study. Moreover, six microbial pathogens are also discussed. The recapitulation of the natural enemies and diseases in DCXC artificial cultivation not only promote the development of integrated pest management of DCXC cultivation but also provide important information to help preserve and develop this valuable resource. PMID:26559703

  5. Soudanones A-G: Antifungal Isochromanones from the Ascomycetous Fungus Cadophora sp. Isolated from an Iron Mine.

    PubMed

    Rusman, Yudi; Held, Benjamin W; Blanchette, Robert A; Wittlin, Sergio; Salomon, Christine E

    2015-06-26

    One new isochromane (pseudoanguillosporin C, 2), seven isochromanones (soudanones A-G, 3-9), and six known analogues including 10 and 11 were isolated from a culture of the fungus Cadophora sp. 10-5-2 M, collected from the subterranean 10th level of the Soudan Underground Iron Mine in Minnesota. All of the compounds were tested against a panel of microbial pathogens, and 2, 3, 10, and 11 were found to have activity against Cryptococcus neoformans (MIC = 35, 40, 20, and 30 μg/mL, respectively). Compound 11 was also active against Candida albicans, with an MIC of 40 μg/mL. PMID:26035018

  6. Wickerhamomyces arborarius f.a., sp. nov., an ascomycetous yeast species found in arboreal habitats on three different continents.

    PubMed

    James, Stephen A; Barriga, Enrique Javier Carvajal; Barahona, Patricia Portero; Harrington, Thomas C; Lee, Ching-Fu; Bond, Christopher J; Roberts, Ian N

    2014-03-01

    Five strains representing a novel yeast species belonging to the genus Wickerhamomyces were independently isolated from Ecuador, Taiwan and the USA. One strain (CLQCA 10-161(T)) was isolated from the white flower of an unidentified plant species collected in the Maquipucuna cloud forest reserve, near Quito, in Ecuador. A second strain (GY7L12) was isolated from the leaf of a Chinese sumac or nutgall tree (Rhus chinensis 'roxburghiana') collected in the Taoyuan mountain area, Kachsiung, in Taiwan. Three additional strains (A543, A546 and A563) were isolated from two species of wood-boring beetle (Xyleborus glabratus and Xyleborinus saxeseni) collected near Clyo, Georgia, USA. Analysis of the D1/D2 domains of the LSU rRNA gene indicated that the novel species belongs to the genus Wickerhamomyces, and is most closely related to Wickerhamomyces sydowiorum, an insect-associated species predominantly found in South Africa. The North American and Taiwanese strains have identical internal transcribed spacer (ITS) sequences and can be distinguished from the Ecuadorian strain based on a single nucleotide substitution in the ITS1 region. The species name of Wickerhamomyces arborarius f.a., sp. nov. is proposed to accommodate these strains, with CLQCA 10-161(T) (?=?CBS 12941(T)?=?NCYC 3743(T)) designated the type strain. PMID:24453230

  7. Production of the alkaloid swainsonine by a fungal endosymbiont of the ascomycete order Chaetothyriales in the host Ipomoea carnea.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Some plant species within the Convolvulaceae (morning glory family) from South America, Africa, and Australia cause a neurologic disease in grazing livestock caused by swainsonine. These convolvulaceous species including Ipomoea carnea contain the indolizidine alkaloid swainsonine, an inhibitor of ...

  8. GzSNF1 Is Required for Normal Sexual and Asexual Development in the Ascomycete Gibberella zeae▿ †

    PubMed Central

    Lee, Seung-Ho; Lee, Jungkwan; Lee, Seunghoon; Park, Eun-Hee; Kim, Ki-Woo; Kim, Myoung-Dong; Yun, Sung-Hwan; Lee, Yin-Won

    2009-01-01

    The sucrose nonfermenting 1 (SNF1) protein kinase of yeast plays a central role in the transcription of glucose-repressible genes in response to glucose starvation. In this study, we deleted an ortholog of SNF1 from Gibberella zeae to characterize its functions by using a gene replacement strategy. The mycelial growth of deletion mutants (ΔGzSNF1) was reduced by 21 to 74% on diverse carbon sources. The virulence of ΔGzSNF1 mutants on barley decreased, and the expression of genes encoding cell-wall-degrading enzymes was reduced. The most distinct phenotypic changes were in sexual and asexual development. ΔGzSNF1 mutants produced 30% fewer perithecia, which matured more slowly, and asci that contained one to eight abnormally shaped ascospores. Mutants in which only the GzSNF1 catalytic domain was deleted had the same phenotype changes as the ΔGzSNF1 strains, but the phenotype was less extreme in the mutants with the regulatory domain deleted. In outcrosses between the ΔGzSNF1 mutants, each perithecium contained ∼70% of the abnormal ascospores, and ∼50% of the asci showed unexpected segregation patterns in a single locus tested. The asexual spores of the ΔGzSNF1 mutants were shorter and had fewer septa than those of the wild-type strain. The germination and nucleation of both ascospores and conidia were delayed in ΔGzSNF1 mutants in comparison with those of the wild-type strain. GzSNF1 expression and localization depended on the developmental stage of the fungus. These results suggest that GzSNF1 is critical for normal sexual and asexual development in addition to virulence and the utilization of alternative carbon sources. PMID:19028993

  9. Candida heliconiae sp. nov., Candida picinguabensis sp. nov. and Candida saopaulonensis sp. nov., three ascomycetous yeasts from Heliconia velloziana (Heliconiaceae).

    PubMed

    Ruivo, Carla C C; Lachance, Marc-André; Rosa, Carlos A; Bacci, Maurício; Pagnocca, Fernando C

    2006-05-01

    Strains belonging to three novel yeast species, Candida heliconiae (four isolates), Candida picinguabensis (three isolates) and Candida saopaulonensis (two isolates), were recovered in the year 2000 from water of flower bracts of Heliconia velloziana L. Emigd. (Heliconiaceae) found in a forest ecosystem site in an Atlantic rainforest of south-eastern Brazil. C. picinguabensis and C. saopaulonensis were nearly identical in morphology and physiology, but sequence divergence in the D1/D2 domain of the large-subunit rDNA indicated that they should be regarded as different species. They belong to the Metschnikowiaceae clade. C. heliconiae had affinities to Pichia mexicana and related species, but was genetically isolated from all currently accepted species in that group. The type strains are C. heliconiae UNESP 00-91C1T (=CBS 10000T=NRRL Y-27813T), C. picinguabensis UNESP 00-89T (=CBS 9999T=NRRL Y-27814T) and C. saopaulonensis UNESP 00-99T (=CBS 10001T=NRRL Y-27815T). PMID:16627669

  10. Wickerhamiella pagnoccae sp. nov. and Candida tocantinsensis sp. nov., two ascomycetous yeasts from flower bracts of Heliconia psittacorum (Heliconiaceae).

    PubMed

    Barbosa, Anne C; Morais, Camila G; Morais, Paula B; Rosa, Luiz H; Pimenta, Raphael S; Lachance, Marc-André; Rosa, Carlos A

    2012-02-01

    Two novel yeast species were isolated from nectar of flower bracts of Heliconia psittacorum (Heliconiaceae) collected in a Cerrado ecosystem in the state of Tocantins, northern Brazil. Wickerhamiella pagnoccae sp. nov., which is closely related to Candida jalapaonensis, is heterothallic and produces one spheroid ascospore per ascus. Candida tocantinsensis sp. nov. belongs to the Metschnikowiaceae clade and its nearest relative is Candida ubatubensis, but the sequence identity (%) in the D1/D2 domains of the rRNA gene is low. The type strain of W. pagnoccae is UFMG-F18C1(T) ( = CBS 12178(T) = NRRL Y-48735(T)) and the type strain of C. tocantinsensis is UFMG-F16D1(T) ( = CBS 12177(T) = NRRL Y-48734(T)). PMID:21478396

  11. Various grain substrates for the production of fruiting bodies and bioactive compounds of the medicinal caterpillar mushroom, Cordyceps militaris (Ascomycetes).

    PubMed

    Liang, Zeng-Chin; Liang, Chih-Hung; Wu, Chiu-Yeh

    2014-01-01

    In this study, several grains such as brown rice (Br), plumule rice (Pr), wheat (W) and pearl barley (Pb) supplemented with 1% (w/w) peptone (P), yeast extract (Ye), ammonia sulfate (As), and monosodium glutamate (Mg) as a nitrogen source, respectively, were used to produce fruiting bodies and bioactive compounds of two strains of Cordyceps militaris. Among these grain substrates, the substrate most suitable to mycelial growth was Pb+Ye for C. militaris H and L. The mushroom strains colonized this substrate in 12.8 and 12.6 days, respectively. For C. militaris L, the fewest days were required for primordial initiation on Br+Ye and Pr+P substrates. The highest yield and biological efficiency was observed with Pb substrate (25.16 g/bottle and 87.36%) and Br+P substrate (21.84 g/bottle and 75.83%) for C. militaris H and L, respectively. In the fruiting bodies of C. militaris H, the highest cordycepin content was cultivated on W+Mg substrate (25.07 mg/g), the highest mannitol content was cultivated with Pr+Mg (153.21 mg/g) and Pr (151.65 mg/g) substrates, and the highest adenosine content was cultivated with Pr+Ye (0.94 mg/g) and Pb+Ye (0.90 mg/g) substrates. In the fruiting bodies of C. militaris L, the highest cordycepin content was cultivated with W+Mg substrate (22.14 mg/g); the highest mannitol content was cultivated with Pb substrate (189.33 mg/g); and the highest adenosine content was cultivated with Pb+Ye substrate (0.71 mg/g). PMID:25404221

  12. An Aspergillus nidulans GH26 endo-β-mannanase with a novel degradation pattern on highly substituted galactomannans.

    PubMed

    von Freiesleben, Pernille; Spodsberg, Nikolaj; Blicher, Thomas Holberg; Anderson, Lars; Jørgensen, Henning; Stålbrand, Henrik; Meyer, Anne S; Krogh, Kristian B R M

    2016-02-01

    The activity and substrate degradation pattern of a novel Aspergillus nidulans GH26 endo-β-mannanase (AnMan26A) was investigated using two galactomannan substrates with varying amounts of galactopyranosyl residues. The AnMan26A was characterized in parallel with the GH26 endomannanase from Podospora anserina (PaMan26A) and three GH5 endomannanases from A. nidulans and Trichoderma reesei (AnMan5A, AnMan5C and TrMan5A). The initial rates and the maximal degree of enzymatically catalyzed conversion of locust bean gum and guar gum galactomannans were determined. The hydrolysis product profile at maximal degree of conversion was determined using DNA sequencer-Assisted Saccharide analysis in High throughput (DASH). This is the first reported use of this method for analyzing galactomannooligosaccharides. AnMan26A and PaMan26A were found to have a novel substrate degradation pattern on the two galactomannan substrates. On the highly substituted guar gum AnMan26A and PaMan26A reached 35-40% as their maximal degree of conversion whereas the three tested GH5 endomannanases only reached 8-10% as their maximal degree of conversion. α-Galactosyl-mannose was identified as the dominant degradation product resulting from AnMan26A and PaMan26A action on guar gum, strongly indicating that these two enzymes can accommodate galactopyranosyl residues in the -1 and in the +1 subsite. The degradation of α-6(4)-6(3)-di-galactosyl-mannopentaose by AnMan26A revealed accommodation of galactopyranosyl residues in the -2, -1 and +1 subsite of the enzyme. Accommodation of galactopyranosyl residues in subsites -2 and +1 has not been observed for other characterized endomannanases to date. Docking analysis of galactomannooligosaccharides in available crystal structures and homology models supported the conclusions drawn from the experimental results. This newly discovered diversity of substrate degradation patterns demonstrates an expanded functionality of fungal endomannanases, than hitherto reported. PMID:26777252

  13. Toll-like receptor 4-interacting SPA4 peptide suppresses the NLRP3 inflammasome in response to LPS and ATP stimuli.

    PubMed

    Ramani, Vijay; Awasthi, Shanjana

    2015-12-01

    Inflammation is induced because of interplay among multiple signaling pathways and molecules during infectious and noninfectious tissue injuries. Crosstalk between Toll-like receptor-4 signaling and the neuronal apoptosis inhibitor protein, major histocompatibility class 2 transcription activator, incompatibility locus protein from Podospora anserina, and telomerase-associated protein (NACHT), leucine-rich repeat (LRR), and pyrin domain-containing protein 3 (NLRP3) inflammasome against pathogen- or damage-associated molecular patterns can cause exaggerated inflammation. We previously established that the Toll-like receptor-4-interacting SPA4 peptide suppresses gram-negative bacterial lipopolysaccharide (Toll-like receptor-4 ligand)-induced nuclear factor-κB and inflammatory response. In the present study, we hypothesized that the SPA4 peptide exerts its anti-inflammatory effects by suppressing the crosstalk between Toll-like receptor-4 signaling and the NLRP3 inflammasome. We evaluated binding of the lipopolysaccharide-ligand to cell-surface Toll-like receptor-4 in the presence or absence of adenosine triphosphate (an NLRP3 inflammasome inducer) by flow cytometry. The expression and activity of NLRP3 inflammasome-related parameters were studied in cells challenged with lipopolysaccharide and adenosine triphosphate using molecular and immunologic methods. The cells were challenged with lipopolysaccharide and treated with SPA4 peptide before (pre-adenosine triphosphate) or after (post-adenosine triphosphate) secondary challenge with adenosine triphosphate. Our data demonstrate that the Toll-like receptor-4-interacting SPA4 peptide does not affect the binding of lipopolysaccharide to Toll-like receptor-4 in the presence or absence of adenosine triphosphate. We also found that the SPA4 peptide inhibits mRNA and cellular protein levels of pro-interleukin-1β and NLRP3, formation of the NLRP3 inflammasome, caspase activity, and release of interleukin-1β. Furthermore, the SPA4 peptide treatment reduced the secreted levels of interleukin-1β from cells overexpressing Toll-like receptor-4 compared with cells expressing the dominant-negative form of Toll-like receptor-4. Together our results suggest that the SPA4 peptide exerts its anti-inflammatory activity by suppressing Toll-like receptor-4-priming of the NLRP3 inflammasome. PMID:26254306

  14. Genomics reveals traces of fungal phenylpropanoid-flavonoid metabolic pathway in the f ilamentous fungus Aspergillus oryzae.

    PubMed

    Juvvadi, Praveen Rao; Seshime, Yasuyo; Kitamoto, Katsuhiko

    2005-12-01

    Fungal secondary metabolites constitute a wide variety of compounds which either play a vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to play a vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites. PMID:16410762

  15. 50 CFR 10.13 - List of Migratory Birds.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ..., American, Recurvirostra americana BEAN-GOOSE, Taiga, Anser fabalis Tundra, Anser serrirostris BEARDLESS..., Fulvous Whistling-Duck Subfamily ANSERINAE Anser fabalis, Taiga Bean-Goose Anser serrirostris, Tundra...

  16. 50 CFR 10.13 - List of Migratory Birds.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ..., American, Recurvirostra americana BEAN-GOOSE, Taiga, Anser fabalis Tundra, Anser serrirostris BEARDLESS..., Fulvous Whistling-Duck Subfamily ANSERINAE Anser fabalis, Taiga Bean-Goose Anser serrirostris, Tundra...

  17. 50 CFR 10.13 - List of Migratory Birds.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., American, Recurvirostra americana BEAN-GOOSE, Taiga, Anser fabalis Tundra, Anser serrirostris BEARDLESS..., Fulvous Whistling-Duck Subfamily ANSERINAE Anser fabalis, Taiga Bean-Goose Anser serrirostris, Tundra...

  18. 50 CFR 10.13 - List of Migratory Birds.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ..., American, Recurvirostra americana BEAN-GOOSE, Taiga, Anser fabalis Tundra, Anser serrirostris BEARDLESS..., Fulvous Whistling-Duck Subfamily ANSERINAE Anser fabalis, Taiga Bean-Goose Anser serrirostris, Tundra...

  19. Mycelial fermentation characteristics and anti-fatigue activities of a Chinese caterpillar fungus, Ophiocordyceps sinensis strain Cs-HK1 (Ascomycetes).

    PubMed

    Wu, Jian-Yong; Leung, Hong-Po; Wang, Wen-Qiang; Xu, Chunping

    2014-01-01

    Mycelial fermentation of an Ophiocordyceps sinensis strain Cs-HK1 was carried out in various volumes of stirred-tank fermenters from 1.6-L and 15-L laboratory scale to 2000-L industrial scale. The mycelial growth in most fermenters had a higher rate, due probably to more efficient oxygen supply, than in shake-flasks. The mycelial fermentation was successfully scaled up to 2000-L industrial fermenters, achieving 30 g/L maximum biomass in 5 days. The Cs-HK1 mycelia formed hairy and fluffy pellets in the fermentation medium and the mycelial broth exhibited pseudoplastic rheology following the power law, with the flow behavior index n decreasing from 0.5 to 0.3, and the flow consistency K and the apparent viscosity µα increasing with time and biomass concentration. The mycelial broth containing biomass and extracellular products harvested from 2000-L fermenters was tested for anti-fatigue activities in forced animal swimming experiments. The mycelium hot water extract showed the most significant effects, increasing the swimming endurance of mice up to 100%, and also increasing the glycogen levels and reducing the lactic acid and blood urea nitrogen levels significantly. The results demonstrated the feasibility of Cs-HK1 mycelial fermentation for large-scale production of bioactive and medicinal materials. PMID:24941032

  20. Cch1 and Mid1 Are Functionally Required for Vegetative Growth under Low-Calcium Conditions in the Phytopathogenic Ascomycete Botrytis cinerea

    PubMed Central

    Harren, Karin

    2013-01-01

    In the filamentous phytopathogen Botrytis cinerea, the Ca2+/calcineurin signaling cascade has been shown to play an important role in fungal growth, differentiation, and virulence. This study deals with the functional characterization of two components of this pathway, the putative calcium channel proteins Cch1 and Mid1. The cch1 and mid1 genes were deleted, and single and double knockout mutants were analyzed during different stages of the fungal life cycle. Our data indicate that Cch1 and Mid1 are functionally required for vegetative growth under conditions of low extracellular calcium, since the growth of both deletion mutants is strongly impaired when they are exposed to the Ca2+-chelating agents EGTA and 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA). The impact of external Ca2+ was investigated by supplementing with CaCl2 and the ionophore A23187, both of which resulted in elevated growth for all mutants. However, deletion of either gene had no impact on germination, sporulation, hyphal morphology, or virulence. By use of the aequorin reporter system to measure intracellular calcium levels, no differences between the mutant strains and the wild type were obtained. Localization studies revealed a subcellular distribution of the Mid1–green fluorescent protein (GFP) fusion protein in network-like filaments, probably the endoplasmic reticulum (ER) membranes, indicating that Mid1 is not a plasma membrane-located calcium channel in B. cinerea. PMID:23475703

  1. The phylogenetic analysis of fungi associated with lichenized ascomycete genus Bryoria reveals new lineages in the Tremellales including a new species Tremella huuskonenii hyperparasitic on Phacopsis huuskonenii.

    PubMed

    Lindgren, Hanna; Diederich, Paul; Goward, Trevor; Myllys, Leena

    2015-09-01

    The basidiomycete order Tremellales includes many species parasitic on or cohabiting with lichen-forming fungi. In this study, we examined the phylogenetic position of Tremellales obtained from Bryoria thalli using nSSU, 5.8S, and partial nLSU sequence data. Both Bayesian and maximum likelihood analyses revealed the presence of basidiomycetous fungi in three separate clades within Tremellales. Tremellales sp. A and Tremella sp. B exist asymptomatically in Bryoria thalli and should thus be regarded as endolichenic rather than lichenicolous fungi. The third lineage represents a new species and is described here as Tremella huuskonenii. It is hyperparasitic over galls induced by Phacopsis huuskonenii, a lichenicolous fungus growing in Bryoria thalli. We also examined the genetic diversity of Tremella sp. B and Tremella huuskonenii with an extended taxon sampling using ITS and partial nLSU sequence data. For comparison, ITS, GAPDH, and Mcm7 regions were used for phylogenetic analyses of the host lichen specimens. According to our results, phylogenetic structure within the two Tremella species does not appear to correlate with the geographic distribution nor with the phylogeny or the secondary chemistry of the host lichen. However, ITS haplotype analysis of T. huuskonenii revealed some genetic differences between European and North American populations as some haplotypes were more common in Europe than in North America and vice versa. PMID:26321732

  2. Starmerella aceti f.a., sp. nov., an ascomycetous yeast species isolated from fungus garden of the leafcutter ant Acromyrmex balzani.

    PubMed

    Melo, Weilan G P; Arcuri, Silvio L; Rodrigues, Andre; Morais, Paula B; Meirelles, Lucas A; Pagnocca, Fernando C

    2014-04-01

    A novel yeast species was recovered from the fungus garden of the leaf-cutting ant Acromyrmex balzani (Hymenoptera: Formicidae). The growth of the novel yeast species is limited by its ability to metabolize only a few carbon and nitrogenous compounds. A remarkable characteristic of this strain is the vigorous growth in 1 % acetic acid. Sequence analysis of the D1/D2 domains of the LSU rRNA gene showed that the novel species belongs to the Starmerella clade and is phenotypically and genetically divergent from currently recognized species in this clade. Described here as Starmerella aceti f.a., sp. nov., it differs by 37 nucleotide substitutions in the D1/D2 region from Starmerella jinningensis CBS 11864(T), the most closely related species. The type strain of Starmerella aceti sp. nov. is TO 125(T) ( = CBMAI 1594(T) = CBS 13086(T)). PMID:24566828

  3. In vitro antibacterial activities of aqueous extracts from Algerian desert truffles (Terfezia and Tirmania, Ascomycetes) against Pseudomonas aeruginosa and Staphylococcus aureus.

    PubMed

    Gouzi, Hicham; Belyagoubi, Larbi; Abdelali, Khadidja Nesrine; Khelifi, Adila

    2011-01-01

    The aim of this study is to evaluate the in vitro effects of aqueous extracts of three species of Algerian desert truffles (Terfezia claveryi, T. leonis, and Tirmania nivea) on the growth of Pseudomonas aeruginosa and Staphylococcus aureus. The antimicrobial activity of the three aqueous extracts was tested using the agar well diffusion and kinetic bacterial growth curves methods. The aqueous extracts of Terfezia claveryi and Tirmania nivea were found to possess a very powerful antibacterial activity against both S. aureus and P. aeruginosa using agar well diffusion. Using 4% and 11% of the aqueous extracts of T. claveryi and T. nivea in the growth medium of S. aureus caused a significant inhibition of S. aureus growth by 86.48% and 99.09%, respectively. The aqueous extracts of T. claveryi and T. nivea were found to cause a significant inhibition of the growth of P. aeruginosa by 71.11% and 100%, respectively. However, the aqueous extract of Terfecia leonis did not show any antibacterial activity. Therefore, T. claveryi and Tirmania nivea can be considered a source of natural therapeutic agents that can be used to treat eye infections caused by resistant bacteria such as P. aeruginosa and S. aureus. PMID:22181843

  4. Cch1 and Mid1 are functionally required for vegetative growth under low-calcium conditions in the phytopathogenic ascomycete Botrytis cinerea.

    PubMed

    Harren, Karin; Tudzynski, Bettina

    2013-05-01

    In the filamentous phytopathogen Botrytis cinerea, the Ca(2+)/calcineurin signaling cascade has been shown to play an important role in fungal growth, differentiation, and virulence. This study deals with the functional characterization of two components of this pathway, the putative calcium channel proteins Cch1 and Mid1. The cch1 and mid1 genes were deleted, and single and double knockout mutants were analyzed during different stages of the fungal life cycle. Our data indicate that Cch1 and Mid1 are functionally required for vegetative growth under conditions of low extracellular calcium, since the growth of both deletion mutants is strongly impaired when they are exposed to the Ca(2+)-chelating agents EGTA and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). The impact of external Ca(2+) was investigated by supplementing with CaCl(2) and the ionophore A23187, both of which resulted in elevated growth for all mutants. However, deletion of either gene had no impact on germination, sporulation, hyphal morphology, or virulence. By use of the aequorin reporter system to measure intracellular calcium levels, no differences between the mutant strains and the wild type were obtained. Localization studies revealed a subcellular distribution of the Mid1-green fluorescent protein (GFP) fusion protein in network-like filaments, probably the endoplasmic reticulum (ER) membranes, indicating that Mid1 is not a plasma membrane-located calcium channel in B. cinerea. PMID:23475703

  5. Mitochondrial Carnitine-Dependent Acetyl Coenzyme A Transport Is Required for Normal Sexual and Asexual Development of the Ascomycete Gibberella zeae

    PubMed Central

    Son, Hokyoung; Min, Kyunghun; Lee, Jungkwan; Choi, Gyung Ja; Kim, Jin-Cheol

    2012-01-01

    Fungi have evolved efficient metabolic mechanisms for the exact temporal (developmental stages) and spatial (organelles) production of acetyl coenzyme A (acetyl-CoA). We previously demonstrated mechanistic roles of several acetyl-CoA synthetic enzymes, namely, ATP citrate lyase and acetyl-CoA synthetases (ACSs), in the plant-pathogenic fungus Gibberella zeae. In this study, we characterized two carnitine acetyltransferases (CATs; CAT1 and CAT2) to obtain a better understanding of the metabolic processes occurring in G. zeae. We found that CAT1 functioned as an alternative source of acetyl-CoA required for lipid accumulation in an ACS1 deletion mutant. Moreover, deletion of CAT1 and/or CAT2 resulted in various defects, including changes to vegetative growth, asexual/sexual development, trichothecene production, and virulence. Although CAT1 is associated primarily with peroxisomal CAT function, mislocalization experiments showed that the role of CAT1 in acetyl-CoA transport between the mitochondria and cytosol is important for sexual and asexual development in G. zeae. Taking these data together, we concluded that G. zeae CATs are responsible for facilitating the exchange of acetyl-CoA across intracellular membranes, particularly between the mitochondria and the cytosol, during various developmental stages. PMID:22798392

  6. Isolation and purification of a polysaccharide from the caterpillar medicinal mushroom Cordyceps militaris (Ascomycetes) fruit bodies and its immunomodulation of RAW 264.7 macrophages.

    PubMed

    Zhu, Lina; Tang, Qingjiu; Zhou, Shuai; Liu, Yanfang; Zhang, Zhong; Gao, Xinhua; Wang, Shiping; Wang, Zhaolong

    2014-01-01

    A novel polysaccharide (CP2-S) was purified from Cordyceps militaris fruit bodies by hot water extraction, ethanol precipitation, DEAE-Sepharose Fast Flow and Sephacryl S-400 high-resolution chromatography. The polysaccharide had a molecular weight of 5.938 × 10(6) g/mol and was mainly composed of glucose. CP2-S had carbohydrate content estimated to be 100% using the phenol-sulfuric acid method. Immunostimulating experiments in vitro indicated that CP2-S could stimulate nitric oxide production, phagocytosis, respiratory burst activity, and secretion of interleukin-1β and interleukin-2 of macrophages, suggesting that this water-soluble polysaccharide from the fruit body of C. militaris is a natural immunostimulating polysaccharide with potential for further application. PMID:24941166

  7. Molecular Markers to Detect the Formation of Heterokaryon and Homokaryon from Asexual Spores of the Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes).

    PubMed

    Wang, Hong; Cai, Tao; Wei, Jing; Feng, Aiping; Lin, Nan; Bao, Dapeng

    2015-01-01

    Cordyceps militaris is widely cultivated on artificial media in China; however, the cultures often are afflicted with the degeneration of nonfruiting strains. To understand the mechanism of degeneration of C. militaris, from the heterokaryotic strain into the homokaryotic strain, we examined the mating-type genes present in individual asexual spores. Further, we determined the distribution ratio of the different mating-type genes among a sample of asexual spores and the growth rate of heterokaryotic and homokaryotic strains of C. militaris. The distribution ratio of 3 groups of asexual spores from C. militaris heterokaryotic strains was determined as 1:1:1 by statistical analysis, whereas that of the two types of nuclei among asexual spores was 1:1. Nearly two-thirds of the asexual spore isolates were homokaryon, which showed a growth speed similar to the heterokaryon. However, the homokaryon (bearing mating-type MAT-HMG) grew significantly faster at times compared with the heterokaryon. Therefore, the purity of the spawn was difficult to establish. C. militaris heterokaryotic strains can transform into a homokaryotic strain following continued subculture. PMID:26756296

  8. Chemical Composition and Medicinal Value of Fruiting Bodies and Submerged Cultured Mycelia of Caterpillar Medicinal Fungus Cordyceps militaris CBS-132098 (Ascomycetes).

    PubMed

    Chan, Jannie Siew Lee; Barseghyan, Gayane S; Asatiani, Mikheil D; Wasser, Solomon P

    2015-01-01

    In this paper, we report the results of a proximate analysis (i.e., moisture, ash, protein, fat, carbohydrates, and energy); a bioactive compounds analysis (i.e., cordycepin and ergothioneine); fatty and amino acid analysis; and analyses of vitamin content, macro- and microelement composition of fruiting body (FB), and mycelial biomass (MB) of medicinal caterpillar fungus Cordyceps militaris strain CBS-132098. These results demonstrate that the FB and MB of C. militaris are good sources of proteins: 59.8% protein content in the FB and 39.5% in the MB. The MB was distinguished by its carbohydrate content (39.6%), which was higher than that of the FB (29.1% carbohydrate). In the FB of C. militaris, the total amino acid content was 57.39 mg/g and in the MB it was 24.98 mg/g. The quantification of the identified fatty acids indicated that palmitic acid, oleic acid, linoleic acid, and linolenic acid were the major fatty acids. The micro- and macroelement compositions were studied. The highest results were calcium (797 mg/kg FB; 11 mg/kg MB); potassium (15,938 mg/kg FB 12,183 mg/kg MB); magnesium (4,227 mg/kg FB; 3,414 mg/kg MB); sodium (171 mg/kg FB; 1,567 mg/kg MB); phosphorus (7,196 mg/kg FB; 14,293 mg/kg MB); and sulfur (5,088 mg/kg FB; 2,558 mg/kg MB). The vitamin composition was studied, and the most abundant vitamins were vitamin A, vitamin B3, and vitamin E. The bioactive components were cordycepin, cordycepic acid (D-mannitol), and ergothioneine. There were differences in cordycepin and ergothioneine contents between the FB and the MB. The cordycepin concentration was 0.11% in the FB and 0.182% in the MB, the cordycepic acid was 4.7 mg/100g in the FB and 5.2 mg/100 g in the MB, and the ergothioneine content was 782.37 mg/kg in the FB and 130.65 mg/kg in the MB. The nutritional values of the FB and the MB of C. militaris detected indicate its potential use in well-balanced diets and sources of bioactive compounds. PMID:26559699

  9. Molecular Cloning and Characterization of a Subtilisin-Like Serine Protease Gene (Pr1) from the Medicinal Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes).

    PubMed

    Shi, Ping; Lu, Zenghui; He, Yuanchuan; Chen, Shijiang; Yan, Jun; Li, Junhong; Zhang, Xiaobing

    2015-01-01

    In the Chinese caterpillar mushroom Ophiocordyceps sinensis, a subtislin-like serine protease (Pr1) is one of the most important enzymes for its infection activity against insect cuticles. The Pr1 gene was isolated from the valuable Chinese medicinal fungus O. sinensis using rapid amplification of 5' and 3' complementary DNA ends. The 2079-bp full-length complementary DNA sequence containing the 1605-bp predicted open reading frame of the Pr1 gene was obtained (GenBank accession no. KF836756). The open reading frame encodes a protein comprising 534 amino acids. Protein sequence multiple alignment analysis revealed high homology with 16 other subtilisin serine proteases and exhibited the highly conserved catalytic domain (D195, H227, and S393). We also constructed a phylogenetic tree in this study. Further molecular studies are needed to elucidate the mechanisms of fungal infection. PMID:26853964

  10. Novel Bioactive Wild Medicinal Mushroom-Xylaria sp. R006 (Ascomycetes) against Multidrug Resistant Human Bacterial Pathogens and Human Cancer Cell Lines.

    PubMed

    Ramesh, Veluchamy; Santosh, Karnewar; Anand, Thangarajan Durai; Shanmugaiah, Vellasamy; Kotamraju, Srigiridhar; Karunakaran, Chandran; Rajendran, Ayyappan

    2015-01-01

    In the present study, the fruiting body extracts of Xylaria sp. strain R006 were obtained from hexane, ethyl acetate and methanol. Among them, the ethyl acetate extract exhibited significant antimicrobial activities against bacterial and fungal pathogens. Based on the effective antimicrobial activity, the crude ethyl acetate extract was fractionized by two-step siliga gel column chromatography. All the fractions were tested for antibacterial activity against drug resistant Staphylococcus aureus strains (1-10) and Pseudomonas aeruginosa strains (1-8). The fraction E showed a maximum inhibition zone of 27.9 mm against drug resistant S. aureus strain 3 and 29.4 mm against drug resistant P. aeruginosa strain 4. Minimal inhibitory concentration of fraction E showed potential result against all the drug resistant strains however, the lowest concentration of 75 µg/mL-1 was observed against S. aureus strains 1 and 6 and P. aeruginosa strain 3. Further, 60 µg/mL of fraction E had significant cytotoxic activity of 54.9, 55.1 and 54.9% against MDA-MB-231 (breast carcinoma cells), A-549 (lung carcinoma cells) and MCF-7 (breast carcinoma cells) human cancer cell lines, respectively. The spectral data revealed that the fraction E has chromophoric groups in it and had the C = O stretching, C-C = C asymmetric stretch, N-H stretch and C-O stretch as functional groups. The results indicate that the metabolites of fruiting bodies of Xylaria sp. R006 are the potential natural source for the development of new anticancer agents. PMID:26756192

  11. SPORE-EXPULSION RATES AND EXTENTS OF BLADE OCCUPATION BY ASCOMYCETES OF THE SMOOTH-CORDGRASS STANDING-DECAY SYSTEM. (R825147)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  12. Structural and functional characterization of the GalNAc/Gal-specific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary.

    PubMed

    Candy, Laure; Van Damme, Els J M; Peumans, Willy J; Menu-Bouaouiche, Laurence; Erard, Monique; Rougé, Pierre

    2003-08-22

    The lectin found in mycelium and sclerotes of the phytopathogenic fungus Sclerotinia sclerotiorum is a homodimer consisting of two identical non-covalently bound subunits of 16,000 Da. CD spectra analysis revealed that the S. sclerotiorum agglutinin (SSA) contains predominantly beta-sheet structures. SSA exhibits specificity towards GalNAc whereby the hydroxyls at positions 4 and 6 of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of SSA can also accommodate disaccharides. The N-terminal sequence of SSA shares no significant similarity with any other protein except a lectin from the Sclerotiniaceae species Ciborinia camelliae. A comparison of SSA and the lectins from C. camelliae and some previously characterized lectins indicates that the Sclerotiniaceae lectins form a homogeneous family of fungal lectins. This newly identified lectin family, which is structurally unrelated to any other family of fungal lectins, is most probably confined to the Ascomycota. PMID:12901882

  13. Phylogenetic analysis of ascomycete yeasts that form coenzyme Q-9 and the proposal of the new genera Babjeviella, Meyerozyma, Millerozyma, Priceomyces and Scheffersomyces

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Species assigned to the genera Debaryomyces, Lodderomyces, Spathaspora and Yamadazyma, as well as selected species of Pichia and Candida that also form coenzyme Q-9, were phylogenetically analyzed from the combined sequences of the D1/D2 domains of the large subunit and the small subunit rRNA genes....

  14. NPS6, Encoding a Non-Ribosomal Peptide Synthetase Involved in Siderophore-Mediated Iron Metabolism, is a Conserved Virulence Determinant of Plant Pathogenic Ascomycetes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    NPS6, encoding a non-ribosomal peptide synthetase, is a virulence determinant in the corn pathogen Cochliobolus heterostrophus and is also involved in resistance to oxidative stress, generated by hydrogen peroxide. Deletion of NPS6 orthologs in the rice pathogen, Cochliobolus miyabeanus, the cereal...

  15. Aging as Evolution-Facilitating Program and a Biochemical Approach to Switch It Off

    NASA Astrophysics Data System (ADS)

    Skulachev, Vladimir P.

    A concept is presented considering aging of living organisms as a final step of their ontogenetic program. It is assumed that such an aging program was invented by biological evolution to facilitate the evolutionary process. Indications are summarized suggesting that controlled production of toxic forms of oxygen (so called reactive oxygen species) by respiring intracellular organelles (mitochondria) is an obligatory component of the aging program. First results of a research project devoted to an attempt to interrupt aging program by antioxidants specifically addressed to mitochondria have been described. Within the framework of the project, antioxidants of a new type (SkQ) were synthesized. SkQs are composed of (i) plastoquinone (an antioxidant moiety), (ii) a penetrating cation, and (iii) a decane or pentane linker. Using planar bilayer phospholipid membranes, we selected SkQ derivatives of the highest penetrability, namely plastoquinonyl decyl triphenylphosphonium (SkQ1), plastoquinonyl decyl rhodamine 19 (SkQR1), and methylplastoquinonyl decyl triphenylphosphonium (SkQ3). Anti- and prooxidant properties of these substances and also of ubiquinonyl-decyl-triphenylphosphonium (MitoQ) were tested in isolated mitochondria. Micromolar concentrations of cationic quinones are found to be very strong prooxidants, but in the lower (sub-micromolar) concentrations they display antioxidant activity which decreases in the series SkQ1 = SkQR1 > SkQ3 > MitoQ. Thus, the window between the anti- and prooxidant effects is the smallest for MitoQ and the largest for SkQ1 and SkQR1. SkQ1 is rapidly reduced by complex III of the mitochondrial respiratory chain, i.e. it is a rechargeable antioxidant. Extremely low concentrations of SkQ1 and SkQR1 completely arrest the H2O2-induced apoptosis in human fibroblasts and HeLa cells (for SkQ1, C 1/2 = 8 · 10-9M). Higher concentrations of SkQ1 are required to block necrosis initiated by reactive oxygen species (ROS). In mice, SkQ1 decelerates the development of three types of accelerated aging (progeria) and also of normal aging, and this effect is especially demonstrative at early stages of aging. The same pattern is shown in invertebrates (Drosophila and Daphnia), and fungus (Podospora anserina). In mammals, the effect of SkQs on aging is accompanied by inhibition of development of such age-related diseases as osteoporosis, involution of thymus, cataract, retinopathy, etc. SkQ1 manifests a strong therapeutic action on some already pronounced retinopathies, in particular, congenital retinal dysplasia. With drops containing 250 nM SkQ1, vision is recovered in 66 of 96 animals (dogs, cats and horses) who became blind because of retinopathy. SkQ1-containing drops instilled into eyes prevent the loss of sight in rabbits suffering from experimental uveitis and restore vision to animals that had already become blind due to this pathology. A favorable effect is also achieved in experimental glaucoma in rabbits. Moreover, the pretreatment of rats with 0.2 nM SkQ1 significantly decreases the H2O2-induced arrhythmia of the isolated heart. SkQ1 strongly reduces the damaged area in myocardial infarction or stroke and prevents the death of animals from kidney infarction. In p53-/- mice, SkQ1 decreases the ROS level in the spleen cells and inhibits appearance of lymphomas which are the main cause of death of such animals. As a result, the lifespan increases. SkQs look like promising drugs to treat aging and age-related diseases.

  16. An attempt to prevent senescence: a mitochondrial approach.

    PubMed

    Skulachev, Vladimir P; Anisimov, Vladimir N; Antonenko, Yuri N; Bakeeva, Lora E; Chernyak, Boris V; Erichev, Valery P; Filenko, Oleg F; Kalinina, Natalya I; Kapelko, Valery I; Kolosova, Natalya G; Kopnin, Boris P; Korshunova, Galina A; Lichinitser, Mikhail R; Obukhova, Lidia A; Pasyukova, Elena G; Pisarenko, Oleg I; Roginsky, Vitaly A; Ruuge, Enno K; Senin, Ivan I; Severina, Inna I; Skulachev, Maxim V; Spivak, Irina M; Tashlitsky, Vadim N; Tkachuk, Vsevolod A; Vyssokikh, Mikhail Yu; Yaguzhinsky, Lev S; Zorov, Dmitry B

    2009-05-01

    Antioxidants specifically addressed to mitochondria have been studied to determine if they can decelerate senescence of organisms. For this purpose, a project has been established with participation of several research groups from Russia and some other countries. This paper summarizes the first results of the project. A new type of compounds (SkQs) comprising plastoquinone (an antioxidant moiety), a penetrating cation, and a decane or pentane linker has been synthesized. Using planar bilayer phospholipid membrane (BLM), we selected SkQ derivatives with the highest permeability, namely plastoquinonyl-decyl-triphenylphosphonium (SkQ1), plastoquinonyl-decyl-rhodamine 19 (SkQR1), and methylplastoquinonyldecyltriphenylphosphonium (SkQ3). Anti- and prooxidant properties of these substances and also of ubiquinonyl-decyl-triphenylphosphonium (MitoQ) were tested in aqueous solution, detergent micelles, liposomes, BLM, isolated mitochondria, and cell cultures. In mitochondria, micromolar cationic quinone derivatives were found to be prooxidants, but at lower (sub-micromolar) concentrations they displayed antioxidant activity that decreases in the series SkQ1=SkQR1>SkQ3>MitoQ. SkQ1 was reduced by mitochondrial respiratory chain, i.e. it is a rechargeable antioxidant. Nanomolar SkQ1 specifically prevented oxidation of mitochondrial cardiolipin. In cell cultures, SkQR1, a fluorescent SkQ derivative, stained only one type of organelles, namely mitochondria. Extremely low concentrations of SkQ1 or SkQR1 arrested H(2)O(2)-induced apoptosis in human fibroblasts and HeLa cells. Higher concentrations of SkQ are required to block necrosis initiated by reactive oxygen species (ROS). In the fungus Podospora anserina, the crustacean Ceriodaphnia affinis, Drosophila, and mice, SkQ1 prolonged lifespan, being especially effective at early and middle stages of aging. In mammals, the effect of SkQs on aging was accompanied by inhibition of development of such age-related diseases and traits as cataract, retinopathy, glaucoma, balding, canities, osteoporosis, involution of the thymus, hypothermia, torpor, peroxidation of lipids and proteins, etc. SkQ1 manifested a strong therapeutic action on some already pronounced retinopathies, in particular, congenital retinal dysplasia. With drops containing 250 nM SkQ1, vision was restored to 67 of 89 animals (dogs, cats, and horses) that became blind because of a retinopathy. Instillation of SkQ1-containing drops prevented the loss of sight in rabbits with experimental uveitis and restored vision to animals that had already become blind. A favorable effect of the same drops was also achieved in experimental glaucoma in rabbits. Moreover, the SkQ1 pretreatment of rats significantly decreased the H(2)O(2) or ischemia-induced arrhythmia of the isolated heart. SkQs strongly reduced the damaged area in myocardial infarction or stroke and prevented the death of animals from kidney ischemia. In p53(-/-) mice, 5 nmol/kgxday SkQ1 decreased the ROS level in the spleen and inhibited appearance of lymphomas to the same degree as million-fold higher concentration of conventional antioxidant NAC. Thus, SkQs look promising as potential tools for treatment of senescence and age-related diseases. PMID:19159610

  17. 50 CFR 10.13 - List of Migratory Birds.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ..., Aethia pygmaea AVOCET, American, Recurvirostra americana BEAN-GOOSE, Taiga, Anser fabalis Tundra, Anser... Taiga, Ficedula albicilla Tufted, Mitrephanes phaeocercus Variegated, Empidonomus varius Vermilion... Subfamily ANSERINAE Anser fabalis, Taiga Bean-Goose Anser serrirostris, Tundra Bean-Goose Anser...

  18. The Wood Rot Ascomycete Xylaria polymorpha Produces a Novel GH78 Glycoside Hydrolase That Exhibits α-l-Rhamnosidase and Feruloyl Esterase Activities and Releases Hydroxycinnamic Acids from Lignocelluloses

    PubMed Central

    Nghi, Do Huu; Bittner, Britta; Kellner, Harald; Jehmlich, Nico; Ullrich, René; Pecyna, Marek J.; Nousiainen, Paula; Sipilä, Jussi; Huong, Le Mai; Hofrichter, Martin

    2012-01-01

    Soft rot (type II) fungi belonging to the family Xylariaceae are known to substantially degrade hardwood by means of their poorly understood lignocellulolytic system, which comprises various hydrolases, including feruloyl esterases and laccase. In the present study, several members of the Xylariaceae were found to exhibit high feruloyl esterase activity during growth on lignocellulosic materials such as wheat straw (up to 1,675 mU g−1) or beech wood (up to 80 mU g−1). Following the ester-cleaving activity toward methyl ferulate, a hydrolase of Xylaria polymorpha was produced in solid-state culture on wheat straw and purified by different steps of anion-exchange and size-exclusion chromatography to apparent homogeneity (specific activity, 2.2 U mg−1). The peptide sequence of the purified protein deduced from the gene sequence and verified by de novo peptide sequencing shows high similarity to putative α-l-rhamnosidase sequences belonging to the glycoside hydrolase family 78 (GH78; classified under EC 3.2.1.40). The purified enzyme (98 kDa by SDS-PAGE, 103 kDa by size-exclusion chromatography; pI 3.7) converted diverse glycosides (e.g., α-l-rhamnopyranoside and α-l-arabinofuranoside) but also natural and synthetic esters (e.g., chlorogenic acid, hydroxycinnamic acid glycoside esters, veratric acid esters, or p-nitrophenyl acetate) and released free hydroxycinnamic acids (ferulic and coumaric acid) from arabinoxylan and milled wheat straw. These catalytic properties strongly suggest that X. polymorpha GH78 is a multifunctional enzyme. It is the first fungal enzyme that combines glycosyl hydrolase with esterase activities and may help this soft rot fungus to degrade lignocelluloses. PMID:22544251

  19. Laccase production by the aquatic ascomycete Phoma sp. UHH 5-1-03 and the white rot basidiomycete Pleurotus ostreatus DSM 1833 during submerged cultivation on banana peels and enzyme applicability for the removal of endocrine-disrupting chemicals.

    PubMed

    Libardi, Nelson; Gern, Regina Maria Miranda; Furlan, Sandra Aparecida; Schlosser, Dietmar

    2012-07-01

    This work aimed to study the production of laccase from Pleurotus ostreatus DSM 1833 and Phoma sp. UHH 5-1-03 using banana peels as alternative carbon source, the subsequent partial purification and characterization of the enzyme, as well the applicability to degrade endocrine disruptors. The laccase stability with pH and temperature, the optimum pH, the K (m) and V(max) parameters, and the molar mass were determined. Tests were conducted for assessing the ability of degradation of the endocrine disruptors t-nonylphenol, bisphenol A, and 17?-ethinylestradiol. Laccase production of 752 and 1,117 U?L? was obtained for Phoma sp. and P. ostreatus, respectively. Phoma sp. laccase showed higher stability with temperature and pH. The laccase from both species showed higher affinity by syringaldazine. The culture broth with banana peels induced the production of two isoforms of P. ostreatus (58.7 and 21 kDa) and one of Phoma sp. laccase (72 kDa). In the first day of incubation, the concentrations of bisphenol A and 17?-ethinylestradiol were reduced to values close to zero and after 3 days the concentration of t-nonylphenol was reduced in 90% by the P. ostreatus laccase, but there was no reduction in its concentration by the Phoma sp. laccase. PMID:22371062

  20. Optimization of solid state culture conditions for the production of adenosine, cordycepin, and D-mannitol in fruiting bodies of medicinal caterpillar fungus Cordyceps militaris (L.:Fr.) Link (Ascomycetes).

    PubMed

    Lim, LekTeng; Lee, ChiaYen; Chang, EngThuan

    2012-01-01

    In general, Cordyceps sinensis is much more popular than C. militaris, though both species contain quite similar bioactive ingredients and exhibit medicinal activities. Many bioactive ingredients have been isolated from C. militaris, such as adenosine, cordycepin, D-mannitol, and exopolysaccharides. C. militaris is claimed to have extensive pharmacological properties, such as: anti-inflammatory; anti-fatigue; anti-bacterial; anti-diabetic; improve lung, liver, and kidney functions; to be beneficial for treating cancer as well as male and female sexual dysfunctions. C. militaris is fast gaining momentum for its so-called health benefits, and it is often used as a substitute for C. sinensis. In view of the growing popularity of C. militaris, nowadays C. militaris cultivation for stroma is also done. There is a great diversity of compounds from different strains of Cordyceps and different artificially cultivated products. This study is to determine the optimum culture parameters integrated with substrate of choice to bring the indoor-cultivated C. militaris to a higher and more consistent level of quality. To achieve the above objective, the resultant products after growth were analyzed for adenosine, cordycepin, and D-mannitol using the high-performance liquid chromatography method. The optimum culture condition to produce a high level of adenosine is by using millet as solid substrate. It must be cultivated in the dark for the first 7 days and harvested on day 40. The optimum culture condition to produce a high level of cordycepin is by using soybean as solid substrate. It must be cultivated in the dark for the first 14 days and harvested on day 50. While a high level of D-mannitol is achieved with millet as the solid substrate. It must be kept in the dark for the first 7 days and harvested on day 50. The adenosine level decreased and cordycepin increased from day 40 of culture to day 50 generally. PMID:22506578

  1. A new species of Jahnulales from Las Ilusiones Lagoon, Tabasco, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During a study on biodiversity of freshwater ascomycetes from an urban tropical lagoon, an ascomycete with similar morphology to species of Jahnulales was obtained. Smooth surface test blocks of Pinus sp., Bucida sp., Cedrela sp. and Tabebuia sp. were submerged in pairs close to a private house whar...

  2. Yippie Yi Yo Mycota Ki Yay! A mycologist’s fervently biased account of how the American western frontier was molded by spores and mycelium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Discussed are white pine blister rust (Cronartium ribicola), cereal rusts (Puccinia spp.), smuts (Tilletia spp.), fungi as agents of recycling in grasslands (e.g., Sporormiella and Podospora spp.), fungal symbionts of bark beetles (e.g., Ophiostoma spp.), impacts of clinical fungi (e.g., Valley Feve...

  3. Relationships among genera of the Saccharomycotina from multigene sequence analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most known species of the subphylum Saccharomycotina (budding ascomycetous yeasts) have now been placed in phylogenetically defined clades following multigene sequence analysis. Terminal clades, which are usually well supported from bootstrap analysis, are viewed as phylogenetically circumscribed ge...

  4. Microsatellite markers for Sclerotinia subarctica nom. prov., a new vegetable pathogen of the High North

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eight polymorphic microsatellite loci were isolated from the ascomycete fungus Sclerotinia subarctica nom. prov. In Alaska, this pathogen causes white mold vegetable diseases sympatrically with the cosmopolitan and closely related Sclerotinia sclerotiorum. Eighteen alleles were observed across the 4...

  5. Genetic structure of the fungal grapevine pathogen Eutypa lata from four continents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The generalist ascomycete fungus Eutypa lata causes Eutypa dieback of grapevine (Vitis vinifera) worldwide. To decipher the cosmopolitan distribution of this fungus, the population genetic structure of 17 geographic samples was investigated from four continental regions (Australia, California, Europ...

  6. Genetic structure of the fungal grapevine pathogen Eutypa lata from four continents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Deciphering the geographic origins of pathogens and elucidating the population biology of these microscopic organisms are necessary steps to establish effective disease-control strategies. The generalist ascomycete fungus Eutypa lata causes Eutypa dieback of grapevine (Vitis vinifera) worldwide. To ...

  7. PHYLOGENETIC DIVERSITY OF MEDICALLY IMPORTANT FUSARIA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to the paucity of phenotypic traits, morphological species recognition greatly underestimates the number of clinically important species of the opportunistic filamentous ascomycete Fusarium. To address this problem, species limits are being investigated using multilocus DNA sequence data, using...

  8. Complex patterns of speciation in cosmopolitan "rock posy" lichens - an integrative approach to discovering and delimiting fungal species in the lichen-forming rhizoplaca melanophthalma speciescomplex

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A growing body of evidence indicates that morphology-based species circumspection of lichenized ascomycetes greatly misrepresents the number of existing species. Recently it has been demonstrated that population-level processes operating within diverging populations can facilitate the identification...

  9. Cytochrome P450 Monooxygenase CYP53 Family in Fungi: Comparative Structural and Evolutionary Analysis and Its Role as a Common Alternative Anti-Fungal Drug Target

    PubMed Central

    Jawallapersand, Poojah; Mashele, Samson Sitheni; Kovačič, Lidija; Stojan, Jure; Komel, Radovan; Pakala, Suresh Babu; Kraševec, Nada; Syed, Khajamohiddin

    2014-01-01

    Cytochrome P450 monooxygenases (CYPs/P450s) are heme-thiolate proteins whose role as a drug target against pathogenic microbes has been explored because of their stereo- and regio-specific oxidation activity. We aimed to assess the CYP53 family's role as a common alternative drug target against animal (including human) and plant pathogenic fungi and its role in fungal-mediated wood degradation. Genome-wide analysis of fungal species revealed the presence of CYP53 members in ascomycetes and basidiomycetes. Basidiomycetes had a higher number of CYP53 members in their genomes than ascomycetes. Only two CYP53 subfamilies were found in ascomycetes and six subfamilies in basidiomycetes, suggesting that during the divergence of phyla ascomycetes lost CYP53 P450s. According to phylogenetic and gene-structure analysis, enrichment of CYP53 P450s in basidiomycetes occurred due to the extensive duplication of CYP53 P450s in their genomes. Numerous amino acids (103) were found to be conserved in the ascomycetes CYP53 P450s, against only seven in basidiomycetes CYP53 P450s. 3D-modelling and active-site cavity mapping data revealed that the ascomycetes CYP53 P450s have a highly conserved protein structure whereby 78% amino acids in the active-site cavity were found to be conserved. Because of this rigid nature of ascomycetes CYP53 P450s' active site cavity, any inhibitor directed against this P450 family can serve as a common anti-fungal drug target, particularly toward pathogenic ascomycetes. The dynamic nature of basidiomycetes CYP53 P450s at a gene and protein level indicates that these P450s are destined to acquire novel functions. Functional analysis of CYP53 P450s strongly supported our hypothesis that the ascomycetes CYP53 P450s ability is limited for detoxification of toxic molecules, whereas basidiomycetes CYP53 P450s play an additional role, i.e. involvement in degradation of wood and its derived components. This study is the first report on genome-wide comparative structural (gene and protein structure-level) and evolutionary analysis of a fungal P450 family. PMID:25222113

  10. Phi Class of Glutathione S-transferase Gene Superfamily Widely Exists in Nonplant Taxonomic Groups

    PubMed Central

    Munyampundu, Jean-Pierre; Xu, You-Ping; Cai, Xin-Zhong

    2016-01-01

    Glutathione S-transferases (GSTs) constitute a superfamily of enzymes involved in detoxification of noxious compounds and protection against oxidative damage. GST class Phi (GSTF), one of the important classes of plant GSTs, has long been considered as plant specific but was recently found in basidiomycete fungi. However, the range of nonplant taxonomic groups containing GSTFs remains unknown. In this study, the distribution and phylogenetic relationships of nonplant GSTFs were investigated. We identified GSTFs in ascomycete fungi, myxobacteria, and protists Naegleria gruberi and Aureococcus anophagefferens. GSTF occurrence in these bacteria and protists correlated with their genome sizes and habitats. While this link was missing across ascomycetes, the distribution and abundance of GSTFs among ascomycete genomes could be associated with their lifestyles to some extent. Sequence comparison, gene structure, and phylogenetic analyses indicated divergence among nonplant GSTFs, suggesting polyphyletic origins during evolution. Furthermore, in silico prediction of functional partners suggested functional diversification among nonplant GSTFs. PMID:26884677

  11. Phi Class of Glutathione S-transferase Gene Superfamily Widely Exists in Nonplant Taxonomic Groups.

    PubMed

    Munyampundu, Jean-Pierre; Xu, You-Ping; Cai, Xin-Zhong

    2016-01-01

    Glutathione S-transferases (GSTs) constitute a superfamily of enzymes involved in detoxification of noxious compounds and protection against oxidative damage. GST class Phi (GSTF), one of the important classes of plant GSTs, has long been considered as plant specific but was recently found in basidiomycete fungi. However, the range of nonplant taxonomic groups containing GSTFs remains unknown. In this study, the distribution and phylogenetic relationships of nonplant GSTFs were investigated. We identified GSTFs in ascomycete fungi, myxobacteria, and protists Naegleria gruberi and Aureococcus anophagefferens. GSTF occurrence in these bacteria and protists correlated with their genome sizes and habitats. While this link was missing across ascomycetes, the distribution and abundance of GSTFs among ascomycete genomes could be associated with their lifestyles to some extent. Sequence comparison, gene structure, and phylogenetic analyses indicated divergence among nonplant GSTFs, suggesting polyphyletic origins during evolution. Furthermore, in silico prediction of functional partners suggested functional diversification among nonplant GSTFs. PMID:26884677

  12. The Genome and Development-Dependent Transcriptomes of Pyronema confluens: A Window into Fungal Evolution

    PubMed Central

    Traeger, Stefanie; Altegoer, Florian; Freitag, Michael; Gabaldon, Toni; Kempken, Frank; Kumar, Abhishek; Marcet-Houben, Marina; Pöggeler, Stefanie; Stajich, Jason E.; Nowrousian, Minou

    2013-01-01

    Fungi are a large group of eukaryotes found in nearly all ecosystems. More than 250 fungal genomes have already been sequenced, greatly improving our understanding of fungal evolution, physiology, and development. However, for the Pezizomycetes, an early-diverging lineage of filamentous ascomycetes, there is so far only one genome available, namely that of the black truffle, Tuber melanosporum, a mycorrhizal species with unusual subterranean fruiting bodies. To help close the sequence gap among basal filamentous ascomycetes, and to allow conclusions about the evolution of fungal development, we sequenced the genome and assayed transcriptomes during development of Pyronema confluens, a saprobic Pezizomycete with a typical apothecium as fruiting body. With a size of 50 Mb and ∼13,400 protein-coding genes, the genome is more characteristic of higher filamentous ascomycetes than the large, repeat-rich truffle genome; however, some typical features are different in the P. confluens lineage, e.g. the genomic environment of the mating type genes that is conserved in higher filamentous ascomycetes, but only partly conserved in P. confluens. On the other hand, P. confluens has a full complement of fungal photoreceptors, and expression studies indicate that light perception might be similar to distantly related ascomycetes and, thus, represent a basic feature of filamentous ascomycetes. Analysis of spliced RNA-seq sequence reads allowed the detection of natural antisense transcripts for 281 genes. The P. confluens genome contains an unusually high number of predicted orphan genes, many of which are upregulated during sexual development, consistent with the idea of rapid evolution of sex-associated genes. Comparative transcriptomics identified the transcription factor gene pro44 that is upregulated during development in P. confluens and the Sordariomycete Sordaria macrospora. The P. confluens pro44 gene (PCON_06721) was used to complement the S. macrospora pro44 deletion mutant, showing functional conservation of this developmental regulator. PMID:24068976

  13. The Complete Mitochondrial Genome of Aix galericulata and Tadorna ferruginea: Bearings on Their Phylogenetic Position in the Anseriformes

    PubMed Central

    Liu, Gang; Zhou, Lizhi; Li, Bo; Zhang, Lili

    2014-01-01

    Aix galericulata and Tadorna ferruginea are two Anatidae species representing different taxonomic groups of Anseriformes. We used a PCR-based method to determine the complete mtDNAs of both species, and estimated phylogenetic trees based on the complete mtDNA alignment of these and 14 other Anseriforme species, to clarify Anseriform phylogenetics. Phylogenetic trees were also estimated using a multiple sequence alignment of three mitochondrial genes (Cyt b, ND2, and COI) from 68 typical species in GenBank, to further clarify the phylogenetic relationships of several groups among the Anseriformes. The new mtDNAs are circular molecules, 16,651 bp (Aix galericulata) and 16,639 bp (Tadorna ferruginea) in length, containing the 37 typical genes, with an identical gene order and arrangement as those of other Anseriformes. Comparing the protein-coding genes among the mtDNAs of 16 Anseriforme species, ATG is generally the start codon, TAA is the most frequent stop codon, one of three, TAA, TAG, and T-, commonly observed. All tRNAs could be folded into canonical cloverleaf secondary structures except for tRNASer (AGY) and tRNALeu (CUN), which are missing the "DHU" arm.Phylogenetic relationships demonstrate that Aix galericula and Tadorna ferruginea are in the same group, the Tadorninae lineage, based on our analyses of complete mtDNAs and combined gene data. Molecular phylogenetic analysis suggests the 68 species of Anseriform birds be divided into three families: Anhimidae, Anatidae, and Anseranatidae. The results suggest Anatidae birds be divided into five subfamilies: Anatinae, Tadorninae, Anserinae, Oxyurinae, and Dendrocygninae. Oxyurinae and Dendrocygninae should not belong to Anserinae, but rather represent independent subfamilies. The Anatinae includes species from the tribes Mergini, Somaterini, Anatini, and Aythyini. The Anserinae includes species from the tribes Anserini and Cygnini. PMID:25375111

  14. The phylogeny of yeasts—A cladistic analysis

    NASA Astrophysics Data System (ADS)

    Sun, Xiuqin; Wu, Baoling

    1992-12-01

    Cladistic analysis was used to clarify the phylogeny of 16 genera of yeasts whose great morphological differences and inclusion in different classification systems resulted in controversies over the taxonomy of seven genera such as Crypeococcus. etc. Some scholars suggest that they belong to Ascomycetes, but others think they belong to fungi imperfecti. After comprehensive cladistic analysis of many genetic characters, the authors consider that the above-mentioned seven genera of yeasts developed in parallel with Ascomycetes so that they should belong to one and the same developmental system.

  15. Altering sexual reproductive mode by interspecific exchange of MAT loci

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sexual fungi can be self-sterile (heterothallic, requiring genetically distinct partners) or selffertile (homothallic, no partner required). In most ascomycetes, a single mating type locus (MAT) controls the ability to reproduce sexually. In the genus Cochliobolus, all heterothallic species have eit...

  16. Influence of host and geographic locale on the distribution of Colletotrichum cereale lineages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Colletotrichum cereale is an ascomycete inhabitant of cool-season grasses of the Pooideae subfamily. The fungus has increased in frequency over the past decade as a destructive pathogen of Poa annua and Agrostis stolonifera cultivated as turfgrass. DNA fingerprinting has revealed two distinct C. c...

  17. Complete Mitochondrial Genome Sequence of the Pezizomycete Pyronema confluens

    PubMed Central

    2016-01-01

    The complete mitochondrial genome of the ascomycete Pyronema confluens has been sequenced. The circular genome has a size of 191 kb and contains 48 protein-coding genes, 26 tRNA genes, and two rRNA genes. Of the protein-coding genes, 14 encode conserved mitochondrial proteins, and 31 encode predicted homing endonuclease genes. PMID:27174271

  18. Entomopathogenic fungi in cornfields and their potential to manage larval western corn rootworm Diabrotica virgifera virgifera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Entomopathogenic Ascomycete fungi are ubiquitous in soil and on phylloplanes, and are important natural enemies of many arthropods, including larval western corn rootworm, Diabrotica virgifera virgifera, which is a major pest of corn. We measured the prevalence of Beauveria bassiana and Metarhizium...

  19. A Review of the Phylogeny and Biology of the Diaporthales

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete order Diaporthales is reviewed based on recent phylogenetic data that outline the families and integrate related asexual fungi. The order is now considered to consist of nine families one of which is newly recognized as the Schizoparmeaceae fam. nov. and two families are recircumscri...

  20. New Phomopsis species identified from wood cankers in eastern North American vineyards.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phomopsis cane and leaf spot, caused by the Ascomycete fungus Phomopsis viticola, is a destructive fruit and foliar disease in eastern North American vineyards. The pathogen typically attacks green tissues, but can also cause wood cankers, presumably due to infection of pruning wounds, as is the cas...

  1. A spatial model for predicting effects of climate change on Swiss needle cast disease severity in the Pacific Northwest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Swiss needle cast disease of Douglas-fir is caused by the ascomycete fungus Phaeocryptopus gaeumannii. Symptoms of the disease are foliage chlorosis and premature needle abscission due to occlusion of stomata by the ascocarps of the pathogen, resulting in impaired needle gas exchange. Severe defol...

  2. Protomyces Unger (1833)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycetous fungal genus Protomyces and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." Species of the genus Protomyces are plant pathogens that attack asters, wild celery, coriander and certain other plants. Symptoms include disruption of stems, lea...

  3. Genetic Diversity of Polyketide Synthase/Nonribosomal Peptide Synthetase Genes in Isolates of the Barley Net Blotch Fungus Pyrenophora teres f. teres

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) are multifunctional enzymes responsible for biosynthesis of diverse small molecules (e.g., mycotoxins and phytotoxins) in filamentous ascomycetes. Both PKS and NRPS genes are present in fungal genomes as large gene families but...

  4. Random T-DNA mutagenesis identifies a Cu-Zn-superoxide dismutase gene as a virulence factor of Sclerotinia sclerotiorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Ascomycetous fungus Sclerotinia sclerotiorum is a devastating pathogen capable of infecting more than 400 plant species including many economically important crops. In order to gain a better mechanistic understanding of its non-specific host-pathogen interactions, random mutagenesis through Agro...

  5. FAMILIES IN THE DIAPORTHALES: A NEW LOOK BASED ON LSU NUCLEAR RDNA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete order Diaporthales includes a number of plant pathogenic fungi, the most notorious of which is Cryphonectria parasitica, the chestnut blight fungus. Relationships among genera in the Diaporthales were evaluated as a basis for the recognition of families and to provide a taxonomic fra...

  6. The potential role of water in spread of conidia of the Neotyphodium endophyte of Poa ampla

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Endophytes of the genus Neotyphodium are mutualistic fungi that colonize many cool season grasses. Neotyphodium endophytes are asexual but related to the ascomycete genus Epichloe. They do not produce obvious structures external to the host and for most of the life cycle are asymptomatic and system...

  7. Intercontinental genetic structure of the fungal grapevine pathogen Eutypa lata

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete fungus Eutypa lata, causal agent of Eutypa dieback of grapevine (Vitis vinifera), impacts all vineyard production systems worldwide. Our objectives were to characterize the population structure of E. lata at different geographical scales to identify migration patterns through ascospor...

  8. Wickerhamiella van der Walt (1973)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycetous yeast genus Wickerhamiella, which has five described species and has been defined from multigene deoxyribonucleic acid (DNA) sequence analysis. The species reproduce by multilateral budding but do not form hyphae or pseudohyphae. Asci typically form a single a...

  9. Wickerhamia Soneda (1960)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Wickerhamia and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus Wickerhamia is known from one species, W. fluorescens. Based on multigene phylogenetic analysis, Wickerhamia appears most closely related to the genera Lodd...

  10. Wickerhamomyces Kurtzman, Robnett & Basehoar-Powers (2008)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Wickerhamomyces and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Wickerhamomyces is newly described and was derived from the genus Pichia following a multigene phylogenetic analysis. At present, there are 17 species...

  11. Back to basics and beyond: increasing the level of resistance to Septoria tritici blotch in wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Septoria tritici blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph: Septoria tritici), is one of the most ubiquitous and important diseases of wheat worldwide. Losses to STB can range from 30 to 50% in disease-conducive climates. Little progress was made in increasing the...

  12. Basis for inhibition of Pyrenophora teres by Laetisaria arvalis, a scanning and transmission electron microscopic study

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The broadly occurring foliar disease of barley, net blotch is caused by Pyrenophora teres, an ascomycete and could result in significant yield loss under heavy disease pressure. The basidiomycete, Laetisaria arvalis has been reported to have biological control activity over some plant pathogens. In ...

  13. Understanding the coevolution of rice blast resistance gene Pi-ta and Magnaporthe oryzae avirulence gene AVR-Pita

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice blast disease caused by the filamentous ascomycetous fungus Magnaporthe oryzae remains to be one of the most serious threats for food security globally. Using resistance (R) genes in integrated cultural practices has been the most powerful practice for rice crop protection. Genetic analysis s...

  14. A ROLE FOR ASCOSPORES IN WHEAT HEAD BLIGHT EPIDEMICS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Ascomycete Gibberella zeae (asexual state Fusarium graminearum) causes serious epidemics of wheat head blight worldwide and contaminates grain with trichothecene mycotoxins that are harmful to human and animal health. Anecdotal evidence dating back to the 19th century indicates that G. zeae asc...

  15. Cryptic Sexuality in Aspergillus parasiticus and A. flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascomycetous fungi of the genus Aspergillus comprise a wide variety of species of biotechnological importance (e.g. A. sojae, A. oryzae, A. niger) as well as pathogens and toxin producers (e.g. A. flavus, A. parasiticus, A. fumigatus, A. nidulans). With the exception of A. nidulans, which is a homot...

  16. Candida Berkhout (1923)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the 314 species assigned to the genus Candida and is to be published in the Yeasts, A Taxonomic Study, 5th edition. Ascomycete yeasts that do not form ascosporic state are assigned to the genus Candida, which is a highly polyphyletic taxonomic form genus. Assigned species in...

  17. Scheffersomyces Kurtzman & M. Suzuki (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Scheffersomyces and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Scheffersomyces has three described species that were earlier assigned to the genus Pichia, but gene sequence analysis showed that the species, now rec...

  18. Meyerozyma Kurtzman & M. Suzuki (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Meyerozyma and is to be published in “The Yeasts, A Taxonomic Study,” 5th edition. The genus Meyerozyma is newly described and was derived from the genus Pichia following a multigene phylogenetic analysis. At present, there are two species assigned...

  19. Millerozyma Kurtzman & M. Suzuki (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Millerozyma and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Millerozyma has two described species that were earlier assigned to the genus Pichia, but gene sequence analysis showed that the species, now reclassified ...

  20. Priceomyuces M. Suzuki & Kurtzman (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Priceomyces and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Priceomyces has five described species that were earlier assigned to the genus Pichia, but gene sequence analysis showed that the species, now reclassified...

  1. Babjeviella Kurtzman & M. Suzuki (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Babjeviella and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus Babjeviella has one known species, B. inositovora, which is represented by three strains, one each from the U.S., Canada and Russia. The genus is phylogenet...

  2. Application of a new approach for characterization and denomination of races of cucurbit powdery mildews – a case study on the Czech pathogen population

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Golovinomyces cichoracearum (Gc) and Podosphaera xanthii (Px) (Ascomycetes, Erysiphaceae) are the most important fungal species causing cucurbit powdery mildew (CPM), a serious disease of field and greenhouse cucurbits. Both species are highly variable, as indicated by the existence of large number ...

  3. Yarrowia van der Walt & von Arx (1980)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This book chapter describes the ascomycete yeast genus Yarrowia and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus has just one described species, Y. lipolytica, and is commonly known by its asexual name Candida lipolytica. The species is widely distributed in name and...

  4. Molecular cloning and characterization of a ToxA-like gene from the maize pathogen Cochliobolus heterostrophus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ToxA, the first discovered fungal proteinaceous host-selective toxin, was originally identified from the tan spot fungus Pyrenophora tritici-repentis (Ptr). Homologues of the PtrToxA gene have not been identified from any other ascomycetes except the leaf/glume blotch fungus Stagonospora nodorum, w...

  5. Effect of levels of wheat residue on the severity of stagonospora nodorum blotch in winter wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stagonospora nodorum blotch (SNB), caused by the ascomycete fungus Stagonospora nodorum, is a major disease of wheat. Wheat residue can be an important source of inoculum, but the effect of different densities of infected debris on disease severity has not been previously determined. Experiments wer...

  6. Description of Groenewaldozyma gen. nov. for placement of Candida auringiensis, Candida salmanticensis and Candida tartarivorans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    DNA sequence analyses have demonstrated that species of the polyphyletic anamorphic ascomycete genus Candida may be members of described teleomorphic genera, members of the Candida tropicalis clade upon which the genus Candida is circumscribed, or members of isolated clades that represent undescribe...

  7. Relationships among genera of the Saccharomycotina (Ascomycota) from multigene phylogenetic analysis of type species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phylogenetic relatedness among ascomycetous yeast genera (subphylum Saccharomycotina, phylum Ascomycota) has been uncertain. In the present study, type species of 70 currently recognized genera are compared from divergence in the nearly entire nuclear gene sequences for large subunit rRNA, small sub...

  8. Characterization and complementation of a fumonisin biosynthetic gene cluster deletion in banana isolates of Fusarium verticillioides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The maize pathogen Fusarium verticillioides produces the fumonisin mycotoxins, which are of significant concern for their animal toxicity. This ascomycete is also reported from banana, but such strains do not produce fumonisins and are not pathogenic on maize seedlings. Southern analysis of several ...

  9. First report of powdery mildew caused by Podosphaera leucotricha on Callery pear in North America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Podosphaera leucotricha (Ellis & Everh.) E.S. Salmon (Ascomycetes, Erysiphales) is the etiological agent of a powdery mildew disease that occurs on rosaceous plants, primarily Malus and Pyrus. This fungus is nearly circumglobal. In May 2009, leaves of Bradford pear (Pyrus calleryana Decne.), some di...

  10. Peterozyma Kurtzman & Robnett (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The chapter describes the ascomycete yeast genus Peterozyma and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Peterozyma is newly described and was derived from the genus Pichia following a multigene phylogenetic analysis. At present, there are two known species, bo...

  11. Zygotorulaspora Kurtzman (2003)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Zygotorulaspora and is to be published in “The Yeasts, A Taxonomic Study, 5th edition.” The two species in this genus were at different times placed in the genera Torulaspora, Zygosaccharomyces and Saccharomyces. Multigene phylogenetic analysis sho...

  12. Blastobotrys von Klopotek (1967)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the anamorphic ascomycete genus Blastobotrys and is to be published in The Yeasts, a Taxonomic Study, 5th edition. The genus Blastobotrys, which represents the asexual state of the genus Trichomonascus, has been phylogenetically defined and has 21 assigned species. Blastobot...

  13. Debaryomyces Lodder & Kreger-van Rij (1952)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Debaryomyces and is to be published in The Yeasts, a Taxonomic Study, 5th edition. The genus Debaryomyces has nine described species, many of which are worldwide in distribution. Most notable species is D. hansenii, which is found in soil, plant de...

  14. CONSERVED REQUIREMENT FOR A PLANT HOST CELL PROTEIN IN POWDERY MILDEW PATHOGENESIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the fungal phylum Ascomycota, the ability to cause disease in plants and animals was gained and lost repeatedly during phylogenesis. In monocotyledonous barley, loss-of-function mlo alleles result in effective immunity against the Ascomycete, Blumeria graminis f. sp. hordei, the causal agent of t...

  15. Saccharomycopsis Schionning (1903)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Saccharomycopsis and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus Saccharomycopsis has 11 assigned species, many worldwide in distribution. Certain species, such as S. fibuligera, are commonly found in fermented, cere...

  16. POPULATION STRUCTURE AND DIVERSITY OF EUTYPA LATA FROM MEDITERRANEAN GRAPE-GROWING REGIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eutypa lata is an ascomycete fungus causing dieback of grape (Vitis vinifera). We examined the genetic structure of eight vineyard collections using nine polymorphic microsatellite loci. In California, isolates were collected from four vineyards (CS1, CS2, M1 and M2) separated by distances of 50 m t...

  17. Cephaloascus Hanawa (1920)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Cephaloascus and is to be published in “The Yeasts, A Taxonomic Study, 5th edition.” The genus Cephaloascus has two species. One, C. albidus, has been isolated from spoiled cranberry pumace, and the second, C. fragrans, is predominantly isolated fr...

  18. Torulaspora Lindner (1904)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Torulaspora and is to be published in "The Yeasts, A Taxonomic Study," 5th edition. The genus Torulaspora is closely related to Zygosaccharomyces and Zygotorulaspora and has six described species. Many of the species are worldwide in distribution. ...

  19. Trigonopsis Schachner emend. Kurtzman & Robnett (2007)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The chapter describes the asexual ascomycete yeast genus Trigonopsis and is to be published in “The Yeasts, A Taxonomic Study, 5th edition.” The genus Trigonopsis has four known species and T. variabilis is famous for producing triangular cells, whereas the other described species do not. Multigen...

  20. Diversity, Pathogenicity And Control of Verticillium Species.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genus Verticillium is a cosmopolitan group of ascomycetous fungi, encompassing phytopathogenic species that cause vascular wilts of plants. Two of these species, V. dahliae and V. albo-atrum, cause billions of dollars in annual crop losses worldwide. The soil habitat of these species, the exte...

  1. The transcription factor FgStuAp influences spore development, pathogenicity, and secondary metabolism in Fusarium graminearum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Members of the APSES family of fungal proteins regulate morphogenesis and virulence in ascomycetes. We deleted the FgStuA gene in Fusarium graminearum and demonstrate its involvement in several different processes. FgStuA is closely related to FoStuA in F. oxysporum and StuA in Aspergillus. Unlike F...

  2. Structural analysis of the inhibition of Pyrenophora teres by Laetisaria arvalis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The basidiomycete fungus, Laetisaria arvalis has been reported to have biological control activity over some plant pathogens [1]. This soil-inhabiting Basidiomycete strain was isolated in a Nebraska sugar beet field [3] . Net blotch, a foliar disease of barley, is caused by the ascomycete fungus Py...

  3. Detection and Identification of Decay Fungi in Spruce Wood by Restriction Fragment Length Polymorphism Analysis of Amplified Genes Encoding rRNA†

    PubMed Central

    Jasalavich, Claudia A.; Ostrofsky, Andrea; Jellison, Jody

    2000-01-01

    We have developed a DNA-based assay to reliably detect brown rot and white rot fungi in wood at different stages of decay. DNA, isolated by a series of CTAB (cetyltrimethylammonium bromide) and organic extractions, was amplified by the PCR using published universal primers and basidiomycete-specific primers derived from ribosomal DNA sequences. We surveyed 14 species of wood-decaying basidiomycetes (brown-rot and white-rot fungi), as well as 25 species of wood-inhabiting ascomycetes (pathogens, endophytes, and saprophytes). DNA was isolated from pure cultures of these fungi and also from spruce wood blocks colonized by individual isolates of wood decay basidiomycetes or wood-inhabiting ascomycetes. The primer pair ITS1-F (specific for higher fungi) and ITS4 (universal primer) amplified the internal transcribed spacer region from both ascomycetes and basidiomycetes from both pure culture and wood, as expected. The primer pair ITS1-F (specific for higher fungi) and ITS4-B (specific for basidiomycetes) was shown to reliably detect the presence of wood decay basidiomycetes in both pure culture and wood; ascomycetes were not detected by this primer pair. We detected the presence of decay fungi in wood by PCR before measurable weight loss had occurred to the wood. Basidiomycetes were identified to the species level by restriction fragment length polymorphisms of the internal transcribed spacer region. PMID:11055916

  4. Mycotoxin production and prevention of aflatoxin contamination in food and feed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aflatoxins are the most prominent group of mycotoxins. They are known to be the most toxic and potent carcinogens naturally produced. They are mainly produced by the ascomycetous fungi Aspergillus flavus and A. parasiticus. Over 40 years of research and investigation has generated a wealth of pub...

  5. Three new anascosporic genera of the Saccharomycotina: Danielozyma gen. nov., Deakozyma gen. nov. and Middelhovenomyces gen. nov.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three new non-ascosporic, ascomycetous yeast genera are proposed based on their isolation from currently described species and genera. Phylogenetic placement of the genera was determined from analysis of nuclear gene sequences for D1/D2 large subunit rRNA, small subunit rRNA, translation elongation...

  6. Occultocarpon, a new monotypic genus of Gnomoniaceae on Alnus nepalensis from China

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new monotypic genus Occultocarpon and its species, O. ailaoshanense, was discovered on the bark of branches of Alnus nepalensis (Betulaceae) in Yunnan, China. A phylogeny based on three genes (LSU, rpb2, tef1-a) reveals that O. ailaoshanense belongs to the Gnomoniaceae (Diaporthales, Ascomycetes) ...

  7. High-Quality Draft Genome Sequence of Candida apicola NRRL Y-50540

    PubMed Central

    Vega-Alvarado, Leticia; Gómez-Angulo, Jorge; Escalante-García, Zazil; Grande, Ricardo; Gschaedler-Mathis, Anne; Amaya-Delgado, Lorena

    2015-01-01

    Candida apicola, a highly osmotolerant ascomycetes yeast, produces sophorolipids (biosurfactants), membrane fatty acids, and enzymes of biotechnological interest. The genome obtained has a high-quality draft for this species and can be used as a reference to perform further analyses, such as differential gene expression in yeast from Candida genera. PMID:26067948

  8. Immune Response of Mormon Crickets that Survived Infection by Beauveria Bassiana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Beauveria bassiana is an entomopathogenic Ascomycete fungus that serves as a biological control agent of Mormon crickets (Anabrus simplex Haldeman) and other grasshopper pests. To measure the dose dependent response of Mormon crickets to fungal attack, we applied B. bassiana strain GHA topically to...

  9. AFLP ANALYSIS OF A WORLDWIDE COLLECTION OF DIDYMELLA BRYONIAE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Didymella bryoniae (anamorph Phoma cucurbitacearum) is an Ascomycete fungus that causes gummy stem blight, a foliar disease that occurs on cucurbits in greenhouses and fields throughout the world. In a previous study using RAPD analysis, little genetic diversity was found among isolates of D. bryon...

  10. MVE1, encoding the velvet gene product homolog in Mycosphaerella graminicola, is associated with aerial mycelium formation, melanin biosynthesis, hyphal swelling, and light signaling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete fungus Mycosphaerella graminicola is an important pathogen of wheat causing economically significant losses. The primary nutritional mode of this fungus is thought to be hemibiotrophic. This pathogenic lifestyle is associated with an early biotrophic stage of nutrient uptake followed ...

  11. MVE1 Encoding the velvet gene product homolog in Mycosphaerella graminicola is associated with aerial mycelium formation, melanin biosynthesis, hyphal swelling, and light signaling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete fungus Mycosphaerella graminicola is an important pathogen of wheat that causes the disease septoria tritici blotch. Despite the serious impact of M. graminicola on wheat production worldwide, knowledge about its molecular biology is limited. The velvet gene, veA, is one of the key re...

  12. Dermatolactone, a cytotoxic fungal sesquiterpene with a novel skeleton.

    PubMed

    Mayer, A; Kpke, B; Anke, H; Sterner, O

    1996-09-01

    The nematicidal 5-pentyl-2-furaldehyde and the cytotoxic sesquiterpene dermatolactone were isolated from the extracts of an Ascomycete belonging to the Dermateaceae. The furan has previously been reported from the Basidiomycete Irpex lacteus, while dermatolactone is a new compound the structure of which was determined by spectrosocpic methods. PMID:8862030

  13. Phylogeny and redescription of Dolabra nepheliae on rambutan and litchi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rambutan (Nephelium lappaceum L.) and lychee (Litchi chinensis Sonn.) are tropical trees in the Sapindaceae that produce delicious edible fruits and are increasingly cultivated in tropical areas. Recently these trees have been afflicted with a stem canker disease caused by the ascomycete Dolabra nep...

  14. Hyphopichia von Arx & van der Walt (1976)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Hyphopichia and is to be published in “The Yeasts, A Taxonomic Study, 5th edition.” The genus Hyphopichia was derived from the genus Pichia and accepted as valid following a multigene phylogenetic analysis. At present, there are two species assigne...

  15. Lachancea Kurtzman (2003)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Lachancea and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Lachancea was recently described following a multigene phylogenetic study and includes species previously assigned to the genera Kluyveromyces, Saccharomyces...

  16. Ogataea Y. Yamada, K. Maeda & Mikata (1994)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The chapter describes the ascomycete yeast genus Ogataea and is to be published in TheYeasts, A Taxonomic Study, 5th edition. The genus Ogataea includes 31 species, many described in the last few years as a result of the availability of species-specific gene sequence databases. All but one of the ...

  17. AGONOMYCETES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Agonomycetes are fungi which usually produce neither sexual (meiotic) nor asexual (mitotic) spores. Some members of this artificial (non-phylogenetic) group are related to ascomycetes, while others are related to basidiomycetes. Many members form spore-like propagules called chlamydospores, papulos...

  18. NRPS4 is responsible for the biosynthesis of destruxins in Metarhizium robertsii ARSEF 2575

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Destruxins (DTXs) are a family of cyclic depsipeptides that include > 35 members produced by Ascomycetous fungi belonging to several different taxa. These metabolites display a plethora of biological activities including toxicity against insects, depolarization of Ca2+ gradient across the plasma mem...

  19. Colletotrichum fungal pathogens and symbionts of ornamental nursery and landscape plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungi in the ascomycete genus Colletotrichum are ranked by the plant pathology community as one of the ten most economically and scientifically important fungal phytopathogens. Major losses due to Colletotrichum are experienced in almost every crop worldwide, including nursery and landscape plants ...

  20. Cryptic Sexuality Influences Aflatoxigenicity in Aspergillus parasiticus and A. flavus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascomycetous fungi of the genus Aspergillus comprise a wide variety of species of biotechnological importance as well as pathogens and toxin producers. Recent studies report A. fumigatus to be heterothallic and possibly undergoing sexual reproduction. We therefore investigated whether compatible mat...

  1. A ToxA-like protein from Cochliobolus heterostrophus induces light-dependent leaf necrosis with host selectivity on maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ToxA, the first discovered fungal proteinaceous host-selective toxin (HST), was originally identified from Pyrenophora tritici-repentis (Ptr), and its homologues have not been identified from any other ascomycetes except Parastagonospora nodorum. Here we report the identification of a ToxA-like pro...

  2. Schwanniomyces Klocker emend. M. Suzuki & Kurtzman (2010)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Schwanniomyces and is to be published in The Yeasts, A Taxonomic Study, 5th edn. The genus Schwanniomyces has seven assigned species, many worldwide in distribution. Schwanniomyces is one of the few yeast genera with species that seem to live in th...

  3. Development of molecular markers for breeding for disease resistant crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice blast disease caused by the filamentous ascomycetes fungus Magnaporthe oryzae and sheath blight disease caused by the soil borne fungus Rhizocotonia solani are the two major rice diseases that threaten stable rice production in the USA and worldwide. These two diseases have been managed with a ...

  4. Sex-linked phenotypic divergence in the hermaphrodite fungus Neurospora tetrasperma

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Here we present a study of the molecular phenotype linked to a large region of suppressed recombination (extending over ~ 7 Mbp and >1,500 genes) surrounding the mating-type (mat) locus of the filamentous ascomycete Neurospora tetrasperma. While the remainder of the genome is largely homoallelic, th...

  5. Species identification of the causal agent of Eutypa dieback of grapevine in northeastern American and southeastern Canadian vineyards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eutypa dieback of Vitis (grape) is caused by the Ascomycete fungus Eutypa lata. The pathogen infects grapevine through wounds, and cause wood canker and dieback symptoms. E. lata has been identified in all major grape production areas in the world. The first report of Eutypa dieback from northeaster...

  6. Ascosphaera subglobosa, a new species from North America associated with the solitary bee Megachile rotundata

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascosphaera is a widespread ascomycetous genus of mostly obligate associates of bees. These fungi have diversified to exploit seemingly every possible substrate available in their bee-associated habitat, occurring as pathogens of the bees, or as saprotrophs on honey, cocoons, nesting materials, poll...

  7. Characterization and complementation of an apparent FUM gene cluster deletion in Fusarium verticillioides.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The filamentous ascomycete Fusarium verticillioides is a worldwide pathogen of maize and produces the fumonisin mycotoxins. Contamination of maize kernels with fumonisin B1 (FB1) is of significant concern because of its causal role in equine leukoencephalomalacia, porcine pulmonary edema, liver and...

  8. A haemagglutinin from the medicinal fungus Cordyceps militaris.

    PubMed

    Wong, Jack H; Wang, Hexiang; Ng, Tzi B

    2009-10-01

    There are only a few reports on agglutinins from ascomycete and medicinal fungi. An HA (haemagglutinin), with an N-terminal amino acid sequence different from those of known lectins, was isolated in the present study from dried fruiting bodies of the medicinal ascomycete fungus Cordyceps militaris. The purification protocol consisted of affinity chromatography, ion-exchange chromatography and gel filtration. The haemagglutinating activity of the HA could not be inhibited by simple sugars or heparin, and was stable over the pH range 2-13 and up to 60 degrees C. Chemical modification of tryptophan and tyrosine residues had no effect. The HA exhibited some antiproliferative activity towards hepatoma (HepG2) cells and inhibited HIV-1 reverse transcriptase (IC50=10 microM). However, it did not exhibit antifungal activity, mitogenic activity towards splenocytes, nitric oxide-inducing activity towards macrophages or RNase activity. The results of the present study add to the meagre information pertaining to agglutinins from ascomycete and medicinal mushrooms. It is revealed in this study that C. militaris HA differs from other ascomycete mushroom HAs in a variety of biochemical characteristics. PMID:19093913

  9. New broad-spectrum resistance to septoria tritici blotch derived from synthetic hexaploid wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Septoria tritici blotch (STB), caused by the ascomycete Mycosphaerella graminicola, is one of the most devastating foliar diseases of wheat. We screened five synthetic hexaploid wheats (SHs), 13 wheat varieties that represent the differential set of cultivars and two susceptible checks with a global...

  10. Kazachstania Zubkova (1971)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Kazachstania and is to be published in "The Yeasts, A Taxonomic Study, 5th edition." The genus Kazachstania is newly described and was constructed from certain species previously assigned to the genera Saccharomyces, Kluyveromyces and Arxozyma follo...

  11. Naumovozyma Kurtzman (2008)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycetous yeast genus Naumovozyma, which was recognized from multigene deoxyribonucleic acid (DNA) sequence analysis. The genus has two describes species, which were formerly classified in the genus Saccharomyces. The species reproduce by multilateral budding but do not...

  12. MOLECULAR MAPPING OF SEPTORIA TRITICI LEAF BLOTCH RESISTANCE IN WHEAT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Septoria tritici leaf blotch (STB), caused by the ascomycete fungus Mycosphaerella graminicola (anamorph Septoria tritici), is one of the most destructive diseases of wheat worldwide. Host plant resistance is the preferred method of disease control. The identification and mapping of STB resistance...

  13. The finished genomic sequence of the Septoria tritici blotch pathogen Mycosphaerella graminicola

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycosphaerella graminicola is the haploid ascomycete that causes Septoria tritici blotch, one of the most important diseases of wheat worldwide. This pathogen is phylogenetically distinct from other fungi that have been sequenced and is hemibiotrophic; early infection is biotrophic, followed by a s...

  14. Mla- and Rom1-mediated control of microRNA398 and chloroplast copper/zinc superoxide dismutase regulates cell death in response to the barley powdery mildew fungus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Barley Mla (Mildew resistance locus a) confers allele-specific interactions with natural variants of the ascomycete fungus, Blumeria graminis f. sp. hordei (Bgh), causal agent of powdery mildew disease. Significant reprogramming of host gene expression occurs upon infection by this obligate biotrop...

  15. Lipomyces Lodder & Kreger-van Rij (1952)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The chapter describes the ascomycete yeast genus Lipomyces and is to be published in “The Yeasts, A Taxonomic Study, 5th edition.” The genus Lipomyces has 16 known species, most of which are isolated exclusively from soil. Cultures of Lipomyces are generally slimy because of the copious production...

  16. Biological and Chemical Complexity of Fusarium proliferatum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The heterothallic ascomycete Fusarium proliferatum (teleomorph Gibberella intermedia) is a genetically diverse biological and phylogenetic species with a worldwide distribution and an unusually broad host range. F. proliferatum is a frequent component of the Fusarium ear rot complexes of maize and ...

  17. Taiwanascus samuelsii sp. nov., an addition to Niessliaceae from the Western Ghats, Kerala, India

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species of Taiwanascus, T. samuelsii, was collected from southern parts of Western Ghats on dead branches of Anacardium occidentale and is described. The new cleistothecial ascomycete is different from the type and only species in Taiwanascus, T. tetrasporus, in cleistothecial size, setae, and...

  18. Molecular phylogeny of Sydowiellaceae, resolving the position of Cainiella

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cainiella is an ascomycete genus associated with arctic alpine plants. The systematic position of Cainiella has been unclear for a long time with current classifications placing the genus in either Sordariales or Xylariales. Our molecular results, based on mtSSU, ITS and nLSU rDNA data, clearly show...

  19. High-Quality Draft Genome Sequence of Candida apicola NRRL Y-50540.

    PubMed

    Vega-Alvarado, Leticia; Gmez-Angulo, Jorge; Escalante-Garca, Zazil; Grande, Ricardo; Gschaedler-Mathis, Anne; Amaya-Delgado, Lorena; Sanchez-Flores, Alejandro; Arrizon, Javier

    2015-01-01

    Candida apicola, a highly osmotolerant ascomycetes yeast, produces sophorolipids (biosurfactants), membrane fatty acids, and enzymes of biotechnological interest. The genome obtained has a high-quality draft for this species and can be used as a reference to perform further analyses, such as differential gene expression in yeast from Candida genera. PMID:26067948

  20. Nakazawaea Y. Yamada, Maeda & Mikata (1994)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the ascomycete yeast genus Nakazawaea and is to be published in The Yeasts, A Taxonomic Study, 5th edition. The genus Nakazawaea was derived from the genus Pichia and its validity was established from a multigene phylogenetic analysis. The genus contains a single species, ...

  1. Real-time PCR detection of the boxwood blight pathogen Calonectria pseudonaviculata

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boxwood blight is a newly emergent, destructive disease of boxwood (genus Buxus), caused by the ascomycete fungus Calonectria pseudonaviculata. Initially identified in Europe in the mid-1990s, the disease was first reported in the U.S. in CT, NC and VA during October 2011. In less than four months, ...

  2. Global genetic structure of the fungal grapevine pathogen Eutypa lata

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ascomycete fungus Eutypa lata is a trunk pathogen of cultivated grapevine (Vitis vinifera) in all major grape-growing regions of the world. Throughout its geographic range, it is considered a generalist pathogen that can complete its life cycle on a broad range of hosts. To decipher the cosmopol...

  3. Plant Clonal Integration Mediates the Horizontal Redistribution of Soil Resources, Benefiting Neighboring Plants.

    PubMed

    Ye, Xue-Hua; Zhang, Ya-Lin; Liu, Zhi-Lan; Gao, Shu-Qin; Song, Yao-Bin; Liu, Feng-Hong; Dong, Ming

    2016-01-01

    Resources such as water taken up by plants can be released into soils through hydraulic redistribution and can also be translocated by clonal integration within a plant clonal network. We hypothesized that the resources from one (donor) microsite could be translocated within a clonal network, released into different (recipient) microsites and subsequently used by neighbor plants in the recipient microsite. To test these hypotheses, we conducted two experiments in which connected and disconnected ramet pairs of Potentilla anserina were grown under both homogeneous and heterogeneous water regimes, with seedlings of Artemisia ordosica as neighbors. The isotopes [(15)N] and deuterium were used to trace the translocation of nitrogen and water, respectively, within the clonal network. The water and nitrogen taken up by P. anserina ramets in the donor microsite were translocated into the connected ramets in the recipient microsites. Most notably, portions of the translocated water and nitrogen were released into the recipient microsite and were used by the neighboring A. ordosica, which increased growth of the neighboring A. ordosica significantly. Therefore, our hypotheses were supported, and plant clonal integration mediated the horizontal hydraulic redistribution of resources, thus benefiting neighboring plants. Such a plant clonal integration-mediated resource redistribution in horizontal space may have substantial effects on the interspecific relations and composition of the community and consequently on ecosystem processes. PMID:26904051

  4. Maintaining heterokaryosis in pseudo-homothallic fungi

    PubMed Central

    Grognet, Pierre; Silar, Philippe

    2015-01-01

    Among all the strategies displayed by fungi to reproduce and propagate, some species have adopted a peculiar behavior called pseudo-homothallism. Pseudo-homothallic fungi are true heterothallics, i.e., they need 2 genetically-compatible partners to mate, but they produce self-fertile mycelium in which the 2 different nuclei carrying the compatible mating types are present. This lifestyle not only enables the fungus to reproduce without finding a compatible partner, but also to cross with any mate it may encounter. However, to be fully functional, pseudo-homothallism requires maintaining heterokaryosis at every stage of the life cycle. We recently showed that neither the structure of the mating-type locus nor hybrid-enhancing effect due to the presence of the 2 mating types accounts for the maintenance of heterokaryosis in the pseudo-homothallic fungus P. anserina. In this addendum, we summarize the mechanisms creating heterokaryosis in P. anserina and 2 other well-known pseudo-homothallic fungi, Neurospora tetrasperma and Agaricus bisporus. We also discuss mechanisms potentially involved in maintaining heterokaryosis in these 3 species. PMID:26479494

  5. Plant Clonal Integration Mediates the Horizontal Redistribution of Soil Resources, Benefiting Neighboring Plants

    PubMed Central

    Ye, Xue-Hua; Zhang, Ya-Lin; Liu, Zhi-Lan; Gao, Shu-Qin; Song, Yao-Bin; Liu, Feng-Hong; Dong, Ming

    2016-01-01

    Resources such as water taken up by plants can be released into soils through hydraulic redistribution and can also be translocated by clonal integration within a plant clonal network. We hypothesized that the resources from one (donor) microsite could be translocated within a clonal network, released into different (recipient) microsites and subsequently used by neighbor plants in the recipient microsite. To test these hypotheses, we conducted two experiments in which connected and disconnected ramet pairs of Potentilla anserina were grown under both homogeneous and heterogeneous water regimes, with seedlings of Artemisia ordosica as neighbors. The isotopes [15N] and deuterium were used to trace the translocation of nitrogen and water, respectively, within the clonal network. The water and nitrogen taken up by P. anserina ramets in the donor microsite were translocated into the connected ramets in the recipient microsites. Most notably, portions of the translocated water and nitrogen were released into the recipient microsite and were used by the neighboring A. ordosica, which increased growth of the neighboring A. ordosica significantly. Therefore, our hypotheses were supported, and plant clonal integration mediated the horizontal hydraulic redistribution of resources, thus benefiting neighboring plants. Such a plant clonal integration-mediated resource redistribution in horizontal space may have substantial effects on the interspecific relations and composition of the community and consequently on ecosystem processes. PMID:26904051

  6. Screening of ecologically diverse fungi for their potential to pretreat lignocellulosic bioenergy feedstock.

    PubMed

    Singh, S; Harms, H; Schlosser, D

    2014-04-01

    A widespread and hitherto by far underexploited potential among ecologically diverse fungi to pretreat wheat straw and digestate from maize silage in the future perspective of using such lignocellulosic feedstock for fermentative bioenergy production was inferred from a screening of nine freshwater ascomycetes, 76 isolates from constructed wetlands, nine peatland isolates and ten basidiomycetes. Wheat straw pretreatment was most efficient with three ascomycetes belonging to the genera Acephala (peatland isolate) and Stachybotrys (constructed wetland isolates) and two white-rot fungi (Hypholoma fasciculare and Stropharia rugosoannulata) as it increased the amounts of water-extractable total sugars by more than 50 % and sometimes up to 150 % above the untreated control. The ascomycetes delignified wheat straw at rates (lignin losses between about 31 and 40 % of the initial content) coming close to those observed with white-rot fungi (about 40 to 57 % lignin removal). Overall, fungal delignification was indicated as a major process facilitating the digestibility of wheat straw. Digestate was generally more resistant to fungal decomposition than wheat straw. Nevertheless, certain ascomycetes delignified this substrate to extents sometimes even exceeding delignification by basidiomycetes. Total sugar amounts of about 20 to 60 % above the control value were obtained with the most efficient fungi (one ascomycete of the genus Phoma, the unspecific wood-rot basidiomycete Agrocybe aegerita and one unidentified constructed wetland isolate). This was accompanied by lignin losses of about 47 to 56 % of the initial content. Overall, digestate delignification was implied to be less decisive for high yields of fermentable sugars than wheat straw delignification. PMID:24504460

  7. Identification and manipulation of the pleuromutilin gene cluster from Clitopilus passeckerianus for increased rapid antibiotic production

    PubMed Central

    Bailey, Andy M.; Alberti, Fabrizio; Kilaru, Sreedhar; Collins, Catherine M.; de Mattos-Shipley, Kate; Hartley, Amanda J.; Hayes, Patrick; Griffin, Alison; Lazarus, Colin M.; Cox, Russell J.; Willis, Christine L.; O’Dwyer, Karen; Spence, David W.; Foster, Gary D.

    2016-01-01

    Semi-synthetic derivatives of the tricyclic diterpene antibiotic pleuromutilin from the basidiomycete Clitopilus passeckerianus are important in combatting bacterial infections in human and veterinary medicine. These compounds belong to the only new class of antibiotics for human applications, with novel mode of action and lack of cross-resistance, representing a class with great potential. Basidiomycete fungi, being dikaryotic, are not generally amenable to strain improvement. We report identification of the seven-gene pleuromutilin gene cluster and verify that using various targeted approaches aimed at increasing antibiotic production in C. passeckerianus, no improvement in yield was achieved. The seven-gene pleuromutilin cluster was reconstructed within Aspergillus oryzae giving production of pleuromutilin in an ascomycete, with a significant increase (2106%) in production. This is the first gene cluster from a basidiomycete to be successfully expressed in an ascomycete, and paves the way for the exploitation of a metabolically rich but traditionally overlooked group of fungi. PMID:27143514

  8. Transport assays in filamentous fungi: kinetic characterization of the UapC purine transporter of Aspergillus nidulans.

    PubMed

    Krypotou, Emilia; Diallinas, George

    2014-02-01

    Transport assays allow the direct kinetic analysis of a specific transporter by measuring apparent Km and Vmax values, and permit the characterization of substrate specificity profiles through competition assays. In this protocol we describe a rapid and easy method for performing uptake assays in the model filamentous ascomycete Aspergillus nidulans. Our method makes use of A. nidulans germinating conidiospores at a defined morphological stage in which most transporters show maximal expression, avoiding technical difficulties associated with the use of mycelia. In combination with the ease of construction of genetic null mutants in A. nidulans, our method allows the rigorous characterization of any transporter in genetic backgrounds that are devoid of other transporters of similar specificity. Here, we use this method to characterize the kinetic parameters and the specificity profile of UapC, a uric acid-xanthine transporter present in all ascomycetes and member of the ubiquitous Nucleobase-Ascorbate Transporter family, in specific genetic backgrounds lacking other relevant transporters. PMID:24355588

  9. A year-round study on functional relationships of airborne fungi with meteorological factors

    NASA Astrophysics Data System (ADS)

    Li, De-Wei; Kendrick, Bryce

    1995-06-01

    Air sampling was conducted in Waterloo, Canada throughout 1992. Functional relationships between aeromycota and meteorological factors were analysed. The meteorological factors were, in descending order of importance: mean temperature, minimum temperature, maximum temperature, mean wind speed, relative humidity (RH), rain, maximum wind speed and snow. The most important airborne fungal propagules in descending order were: total fungal spores, unidentified Ascomycetes, Cladosporium, Coprinus, unidentified Basidiomycetes, Alternaria and unidentified fungi. Most airborne fungal taxa had highly significant relationship with temperature, but Aspergillus/Penicillium, hyphal fragments and Epicoccum did not. Epicoccum and hyphal fragments were positively associated with wind speed. In comparison with other airborne fungal taxa, Leptosphaeria and unidentified Ascomycetes were more closely correlated with rain and RH during the growing season.

  10. Genomic perspectives on the evolution of fungal entomopathogenicity in Beauveria bassiana

    PubMed Central

    Xiao, Guohua; Ying, Sheng-Hua; Zheng, Peng; Wang, Zheng-Liang; Zhang, Siwei; Xie, Xue-Qin; Shang, Yanfang; St. Leger, Raymond J.; Zhao, Guo-Ping; Wang, Chengshu; Feng, Ming-Guang

    2012-01-01

    The ascomycete fungus Beauveria bassiana is a pathogen of hundreds of insect species and is commercially produced as an environmentally friendly mycoinsecticide. We sequenced the genome of B. bassiana and a phylogenomic analysis confirmed that ascomycete entomopathogenicity is polyphyletic, but also revealed convergent evolution to insect pathogenicity. We also found many species-specific virulence genes and gene family expansions and contractions that correlate with host ranges and pathogenic strategies. These include B. bassiana having many more bacterial-like toxins (suggesting an unsuspected potential for oral toxicity) and effector-type proteins. The genome also revealed that B. bassiana resembles the closely related Cordyceps militaris in being heterothallic, although its sexual stage is rarely observed. A high throughput RNA-seq transcriptomic analysis revealed that B. bassiana could sense and adapt to different environmental niches by activating well-defined gene sets. The information from this study will facilitate further development of B. bassiana as a cost-effective mycoinsecticide. PMID:22761991

  11. Dolabra nepheliae on rambutan and lychee represents a novel lineage of phytopathogenic Eurotiomycetes

    PubMed Central

    Schoch, Conrad L.; Farr, David F.; Nishijima, Kate; Keith, Lisa; Goenaga, Ricardo

    2010-01-01

    Rambutan (Nephelium lappaceum) and lychee (Litchi chinensis) are tropical trees in the Sapindaceae that produce delicious edible fruits and are increasingly cultivated in tropical regions. These trees are afflicted with a stem canker disease associated with the ascomycete Dolabra nepheliae. Previously known from Asia and Australia, this fungus was recently reported from Hawaii and Puerto Rico. The sexual and asexual states of Dolabra nepheliae are redescribed and illustrated. In addition, the ITS and large subunit of the nuclear ribosomal DNA plus fragments from the genes RPB2, TEF1, and the mitochondrial small ribosomal subunit were sequenced for three isolates of D. nepheliae and compared with other sequences of ascomycetes. It was determined that D. nepheliae represents a new lineage within the Eurotiomycetes allied with Phaeomoniella chlamydospora, the causal agent of Petri grapevine decline. PMID:20802819

  12. Identification and manipulation of the pleuromutilin gene cluster from Clitopilus passeckerianus for increased rapid antibiotic production.

    PubMed

    Bailey, Andy M; Alberti, Fabrizio; Kilaru, Sreedhar; Collins, Catherine M; de Mattos-Shipley, Kate; Hartley, Amanda J; Hayes, Patrick; Griffin, Alison; Lazarus, Colin M; Cox, Russell J; Willis, Christine L; O'Dwyer, Karen; Spence, David W; Foster, Gary D

    2016-01-01

    Semi-synthetic derivatives of the tricyclic diterpene antibiotic pleuromutilin from the basidiomycete Clitopilus passeckerianus are important in combatting bacterial infections in human and veterinary medicine. These compounds belong to the only new class of antibiotics for human applications, with novel mode of action and lack of cross-resistance, representing a class with great potential. Basidiomycete fungi, being dikaryotic, are not generally amenable to strain improvement. We report identification of the seven-gene pleuromutilin gene cluster and verify that using various targeted approaches aimed at increasing antibiotic production in C. passeckerianus, no improvement in yield was achieved. The seven-gene pleuromutilin cluster was reconstructed within Aspergillus oryzae giving production of pleuromutilin in an ascomycete, with a significant increase (2106%) in production. This is the first gene cluster from a basidiomycete to be successfully expressed in an ascomycete, and paves the way for the exploitation of a metabolically rich but traditionally overlooked group of fungi. PMID:27143514

  13. Molecular characterisation of fungal endophytic morphospecies associated with the indigenous forest tree, Theobroma gileri, in Ecuador.

    PubMed

    Thomas, Sarah E; Crozier, Jayne; Catherine Aime, M; Evans, Harry C; Holmes, Keith A

    2008-07-01

    Fungal endophytes were isolated from healthy stems and pods of Theobroma gileri, an alternative host of the frosty pod rot pathogen of cacao. Non-sporulating isolates were grouped into 46 different morphological species according to their colony morphology. Many of these morphospecies were assumed to be basidiomycetes and, therefore, were of particular interest. Basidiomycetous endophytes have received far less attention than ascomycetes and also have potential as biological control agents of the basidiomycetous pathogens of T. cacao: Moniliophthora roreri (frosty pod rot pathogen) and M. perniciosa (witches' broom disease). The morphospecies were further characterised by molecular analyses. Amplification of the nuLSU was undertaken for phylogenetic placement of these non-sporulating cultures and revealed a total of 31 different taxa of which 15 were basidiomycetes belonging to the class Agaricomycetes, and 16 ascomycetes primarily belonging to the Sordariomycetes. PMID:18534836

  14. Assimilation of Unusual Carbon Compounds

    NASA Astrophysics Data System (ADS)

    Middelhoven, Wouter J.

    Yeast taxa traditionally are distinguished by growth tests on several sugars and organic acids. During the last decades it became apparent that many yeast species assimilate a much greater variety of naturally occurring carbon compounds as sole source of carbon and energy. These abilities are indicative of a greater role of yeasts in the carbon cycle than previously assumed. Especially in acidic soils and other habitats, yeasts may play a role in the degradation of carbon compounds. Such compounds include purines like uric acid and adenine, aliphatic amines, diamines and hydroxyamines, phenolics and other benzene compounds and polysaccharides. Assimilation of purines and amines is a feature of many ascomycetes and basidiomycetes. However, benzene compounds are degraded by only a few ascomycetous yeasts (e.g. the Stephanoascus/ Blastobotrys clade and black yeastlike fungi) but by many basidiomycetes, e.g. Filobasidiales, Trichosporonales, red yeasts producing ballistoconidia and related species, but not by Tremellales. Assimilation of polysaccharides is wide-spread among basidiomycetes

  15. Nucleotide Composition of Nucleic Acids of Fungi II. Deoxyribonucleic Acids

    PubMed Central

    Storck, Roger

    1966-01-01

    Storck, Roger (The University of Texas, Austin). Nucleotide composition of nucleic acids from fungi. II. Deoxyribonucleic acids. J. Bacteriol. 91:227–230. 1966.—The nucleotide composition of the deoxyribonucleic acids (DNA) present in extracts of 30 species of fungi was determined. The results were analyzed, together with those in the literature. It was found that the content, in moles per cent of guanine plus cytosine (GC content), varied from 38 to 63% in a distribution composed of 9 species of zygomycetes, 14 of ascomycetes, and 9 each of deuteromycetes and basidiomycetes. The GC content ranges were: 38 to 48% for the zygomycetes, 38 to 54% for the ascomycetes, 47 to 62% for the deuteromycetes, and 44 to 63% for the basidiomycetes. The GC content ranged from 38 to 40% for four Mucor species. The base composition of fungal DNA appears, therefore, to have a taxonomic and phylogenetic significance. PMID:5948191

  16. Dolabra nepheliae on rambutan and lychee represents a novel lineage of phytopathogenic Eurotiomycetes.

    PubMed

    Rossman, Amy Y; Schoch, Conrad L; Farr, David F; Nishijima, Kate; Keith, Lisa; Goenaga, Ricardo

    2010-07-01

    Rambutan (Nephelium lappaceum) and lychee (Litchi chinensis) are tropical trees in the Sapindaceae that produce delicious edible fruits and are increasingly cultivated in tropical regions. These trees are afflicted with a stem canker disease associated with the ascomycete Dolabra nepheliae. Previously known from Asia and Australia, this fungus was recently reported from Hawaii and Puerto Rico. The sexual and asexual states of Dolabra nepheliae are redescribed and illustrated. In addition, the ITS and large subunit of the nuclear ribosomal DNA plus fragments from the genes RPB2, TEF1, and the mitochondrial small ribosomal subunit were sequenced for three isolates of D. nepheliae and compared with other sequences of ascomycetes. It was determined that D. nepheliae represents a new lineage within the Eurotiomycetes allied with Phaeomoniella chlamydospora, the causal agent of Petri grapevine decline. PMID:20802819

  17. NADH dehydrogenase subunit genes in the mitochondrial DNA of yeasts.

    PubMed Central

    Nosek, J; Fukuhara, H

    1994-01-01

    The genes encoding the NADH dehydrogenase subunits of respiratory complex I have not been identified so far in the mitochondrial DNA (mtDNA) of yeasts. In the linear mtDNA of Candida parapsilosis, we found six new open reading frames whose sequences were unambiguously homologous to those of the genes known to code for NADH dehydrogenase subunit proteins of different organisms, i.e., ND1, ND2, ND3, ND4L, ND5, and ND6. The gene for ND4 also appears to be present, as judged from hybridization experiments with a Podospora gene probe. Specific transcripts from these open reading frames (ND genes) could be detected in the mitochondria. Hybridization experiments using C. parapsilosis genes as probes suggested that ND genes are present in the mtDNAs of a wide range of yeast species including Candida catenulata, Pichia guilliermondii, Clavispora lusitaniae, Debaryomyces hansenii, Hansenula polymorpha, and others. Images PMID:7521869

  18. Phylogenetic studies of two Anas platyrhynchos (Anatini: Anatinae) in Hunan province of China based on complete mitochondrial DNA sequences.

    PubMed

    He, Xi; Lin, Qian; Cao, Rong; Yuan, Ya-Ting; Pan, Di-Zi; Yun, Long; Zhang, Shi-Rui; Hou, De-Xing

    2016-07-01

    In this study, we cloned and sequenced the complete mitochondrial DNAs of Chinese duck, Anas platyrhynchos, population from two different areas of Hunan province in China. The Anas platyrhynchos breed Linwu duck (LW) sample was taken from the Linwu county of Chenzhou city, and the Anas platyrhynchos breed Youxian duck (YX) sample was taken from the Youxian county of Zhuzhou city. The lengths of their complete mitochondrial genome were 16,604 bp (LW) and 16,606 bp (YX), respectively. The organization of the two Anas platyrhynchos breed mitochondrial genomes was similar to those reported from other duck mitochondrial genomes. Phylogenetic analyses using N-J computational algorithms showed that the analyzed species are divided into four major clades: Anatinae, Anserinae, Dendrocygninae and Anseranatidae. Also, the Linwu duck and Youxian duck have highly similar phylogenetic relationship. PMID:26057008

  19. Isolation of Hanseniaspora uvarum (Kloeckera apiculata) in humans.

    PubMed

    García-Martos, P; Hernández-Molina, J M; Galán, F; Ruiz-Henestrosa, J R; García-Agudo, R; Palomo, M J; Mira, J

    Isolation of Hanseniaspora uvarum, a yeast of the ascomycetes group, whose anamorph corresponds to Kloeckera apiculata, obtained from stool and two ungual specimens from three patients, is reported. This yeast has been found in soil, water, various fruits, bivalve molluscs, crabs, prawns and fruit flies; in Spain, it has been described in the fermentation processes of some wines. In our region, it has also been found in the intestine of mackerel (Scomber scombrus). Its finding in humans constitutes a clinical rarity. PMID:10481287

  20. Release of Dimethylsulfide from Dimethylsulfoniopropionate by Plant-Associated Salt Marsh Fungi

    PubMed Central

    Bacic, M. K.; Newell, S. Y.; Yoch, D. C.

    1998-01-01

    The range of types of microbes with dimethylsulfoniopropionate (DMSP) lyase capability (enzymatic release of dimethylsulfide [DMS] from DMSP) has recently been expanded from bacteria and eukaryotic algae to include fungi (a species of the genus Fusarium [M. K. Bacic and D. C. Yoch, Appl. Environ. Microbiol. 64:106–111, 1998]). Fungi (especially ascomycetes) are the predominant decomposers of shoots of smooth cordgrass, the principal grass of Atlantic salt marshes of the United States. Since the high rates of release of DMS from smooth cordgrass marshes have a temporal peak that coincides with peak shoot death, we hypothesized that cordgrass fungi were involved in this DMS release. We tested seven species of the known smooth cordgrass ascomycetes and discovered that six of them exhibited DMSP lyase activity. We also tested two species of ascomycetes from other DMSP-containing plants, and both were DMSP lyase competent. For comparison, we tested 11 species of ascomycetes and mitosporic fungi from halophytes that do not contain DMSP; of these 11, only 3 were positive for DMSP lyase. A third group tested, marine oomycotes (four species of the genera Halophytophthora and Pythium, mostly from mangroves), showed no DMSP lyase activity. Two of the strains of fungi found to be positive for DMSP lyase also exhibited uptake of DMS, an apparently rare combination of capabilities. In conclusion, a strong correlation exists between a fungal decomposer’s ability to catabolize DMSP via the DMSP lyase pathway and the host plant’s production of DMSP as a secondary product. PMID:16349548

  1. Peritonitis Due to Thermoascus taitungiacus (Anamorph Paecilomyces taitungiacus)

    PubMed Central

    Korzets, Asher; Weinberger, Miriam; Chagnac, Avry; Goldschmied-Reouven, Anna; Rinaldi, Michael G.; Sutton, Deanna A.

    2001-01-01

    The first case of human disease due to the thermophilic ascomycete Thermoascus taitungiacus (the teleomorph of Paecilomyces taitungiacus) is presented. T. taitungiacus was recovered from four dialysate fluid specimens of a 57-year-old patient undergoing chronic peritoneal dialysis. Identification was based upon cylindrical conidia, reddish orange nonostiolate ascomata, lack of growth at 20°C, thermotolerance, and ascospores that appeared pale yellow, elliptical, thick walled, and predominately echinulate by light microscopy but irregularly verrucose by scanning electron microscopy. PMID:11158134

  2. Structure Analysis Uncovers a Highly Diverse but Structurally Conserved Effector Family in Phytopathogenic Fungi.

    PubMed

    de Guillen, Karine; Ortiz-Vallejo, Diana; Gracy, Jérome; Fournier, Elisabeth; Kroj, Thomas; Padilla, André

    2015-10-01

    Phytopathogenic ascomycete fungi possess huge effector repertoires that are dominated by hundreds of sequence-unrelated small secreted proteins. The molecular function of these effectors and the evolutionary mechanisms that generate this tremendous number of singleton genes are largely unknown. To get a deeper understanding of fungal effectors, we determined by NMR spectroscopy the 3-dimensional structures of the Magnaporthe oryzae effectors AVR1-CO39 and AVR-Pia. Despite a lack of sequence similarity, both proteins have very similar 6 β-sandwich structures that are stabilized in both cases by a disulfide bridge between 2 conserved cysteins located in similar positions of the proteins. Structural similarity searches revealed that AvrPiz-t, another effector from M. oryzae, and ToxB, an effector of the wheat tan spot pathogen Pyrenophora tritici-repentis have the same structures suggesting the existence of a family of sequence-unrelated but structurally conserved fungal effectors that we named MAX-effectors (Magnaporthe Avrs and ToxB like). Structure-informed pattern searches strengthened this hypothesis by identifying MAX-effector candidates in a broad range of ascomycete phytopathogens. Strong expansion of the MAX-effector family was detected in M. oryzae and M. grisea where they seem to be particularly important since they account for 5-10% of the effector repertoire and 50% of the cloned avirulence effectors. Expression analysis indicated that the majority of M. oryzae MAX-effectors are expressed specifically during early infection suggesting important functions during biotrophic host colonization. We hypothesize that the scenario observed for MAX-effectors can serve as a paradigm for ascomycete effector diversity and that the enormous number of sequence-unrelated ascomycete effectors may in fact belong to a restricted set of structurally conserved effector families. PMID:26506000

  3. Camarosporium arezzoensis on Cytisus sp., an addition to sexual state of Camarosporium sensu stricto

    PubMed Central

    Tibpromma, Saowaluck; Wijayawardene, Nalin N.; Manamgoda, Dimuthu S.; Boonmee, Saranyaphat; Wanasinghe, Dhanushka N.; Camporesi, Erio; Yang, Jun-Bo; Hyde, Kevin D.

    2015-01-01

    During a study of saprobic fungi from Bagno di Cetica Province, Italy, we collected a pleosporoid ascomycete on stems of Cytisus sp. In morphology, our collection is similar to Cucurbitaria species, but molecular analysis of SSU, LSU and ITS genes reveals it can be referred to Camarosporium. In this study we compare all other Cucurbitaria species from Cytisus sp. and based on both morphology and molecular data, we introduce our collection as a new species in Camarosporium viz. C. arezzoensis. PMID:26858532

  4. Medicinal mushrooms: Towards a new horizon

    PubMed Central

    Ganeshpurkar, A.; Rai, G.; Jain, A. P.

    2010-01-01

    The arising awareness about functional food has created a boom in this new millennium. Mushrooms are widely consumed by the people due to their nutritive and medicinal properties. Belonging to taxonomic category of basidiomycetes or ascomycetes, these mushrooms possess antioxidant and antimicrobial properties. They are also one of the richest source of anticancer and immunomodulating agents. Thus these novel myochemicals from these mushrooms are the wave of future. PMID:22228952

  5. Slippery Scar: A New Mushroom Disease in Auricularia polytricha

    PubMed Central

    Sun, Jie

    2012-01-01

    A new disease, the slippery scar, was investigated in cultivated bags of Auricularia polytricha. This fungus was isolated from the infected mycelia of cultivated bags. Based on morphological observation, rDNA-internal transcribed spacer and 18S sequence analysis, this pathogen was identified as the Ascomycete Scytalidium lignicola. According to Koch's Postulation, the pathogenicity of S. lignicola to the mycelia of A. polytricha was confirmed. The parasitism of this fungus on mushroom mycelia in China has not been reported before. PMID:22870056

  6. Structure Analysis Uncovers a Highly Diverse but Structurally Conserved Effector Family in Phytopathogenic Fungi

    PubMed Central

    Gracy, Jérome; Fournier, Elisabeth; Kroj, Thomas; Padilla, André

    2015-01-01

    Phytopathogenic ascomycete fungi possess huge effector repertoires that are dominated by hundreds of sequence-unrelated small secreted proteins. The molecular function of these effectors and the evolutionary mechanisms that generate this tremendous number of singleton genes are largely unknown. To get a deeper understanding of fungal effectors, we determined by NMR spectroscopy the 3-dimensional structures of the Magnaporthe oryzae effectors AVR1-CO39 and AVR-Pia. Despite a lack of sequence similarity, both proteins have very similar 6 β-sandwich structures that are stabilized in both cases by a disulfide bridge between 2 conserved cysteins located in similar positions of the proteins. Structural similarity searches revealed that AvrPiz-t, another effector from M. oryzae, and ToxB, an effector of the wheat tan spot pathogen Pyrenophora tritici-repentis have the same structures suggesting the existence of a family of sequence-unrelated but structurally conserved fungal effectors that we named MAX-effectors (Magnaporthe Avrs and ToxB like). Structure-informed pattern searches strengthened this hypothesis by identifying MAX-effector candidates in a broad range of ascomycete phytopathogens. Strong expansion of the MAX-effector family was detected in M. oryzae and M. grisea where they seem to be particularly important since they account for 5–10% of the effector repertoire and 50% of the cloned avirulence effectors. Expression analysis indicated that the majority of M. oryzae MAX-effectors are expressed specifically during early infection suggesting important functions during biotrophic host colonization. We hypothesize that the scenario observed for MAX-effectors can serve as a paradigm for ascomycete effector diversity and that the enormous number of sequence-unrelated ascomycete effectors may in fact belong to a restricted set of structurally conserved effector families. PMID:26506000

  7. Tubulin modulating antifungal and antiproliferative pyrazinone derivatives.

    PubMed

    Taggi, Andrew E; Stevenson, Thomas M; Bereznak, James F; Sharpe, Paula L; Gutteridge, Steven; Forman, Robert; Bisaha, John J; Cordova, Daniel; Crompton, Martina; Geist, Lora; Kovacs, Patrick; Marshall, Eric; Sheth, Ritesh; Stavis, Courtney; Tseng, Chi-Ping

    2016-02-01

    A novel class of synthetic tubulin polymerization disruptors, based on a substituted pyrazin-2-one core, has been discovered. These molecules have proven to be potent broad spectrum fungicides, with activity on agriculturally important ascomycete and basidiomycete pathogens. They have also been found to be particularly potent against human rhabdomyosarcoma cells. Using an efficient synthetic route, the agricultural and medicinal activity was explored. PMID:26386818

  8. Genome Sequence of the Food Spoilage Yeast Zygosaccharomyces bailii CLIB 213T

    PubMed Central

    Bigey, Frédéric; Devillers, Hugo; Neuvéglise, Cécile; Dequin, Sylvie

    2013-01-01

    The ascomycetous yeast Zygosaccharomyces bailii is one of the most problematic spoilage yeasts in food and beverage industries, due to its exceptional resistance to various stresses. A better understanding of the molecular mechanisms underlying these stress resistance phenotypes might help develop strategies to improve food quality. Thus, we determined and annotated the genome sequence of the strain Z. bailii CLIB 213T (= CBS 680). PMID:23969048

  9. Spider, bacterial and fungal phospholipase D toxins make cyclic phosphate products.

    PubMed

    Lajoie, Daniel M; Cordes, Matthew H J

    2015-12-15

    Phospholipase D (PLD) toxins from sicariid spiders, which cause disease in mammals, were recently found to convert their primary substrates, sphingomyelin and lysophosphatidylcholine, to cyclic phospholipids. Here we show that two PLD toxins from pathogenic actinobacteria and ascomycete fungi, which share distant homology with the spider toxins, also generate cyclic phospholipids. This shared function supports divergent evolution of the PLD toxins from a common ancestor and suggests the importance of cyclic phospholipids in pathogenicity. PMID:26482933

  10. Draft genome sequence of the dimorphic yeast Yarrowia lipolytica, strain W29

    SciTech Connect

    Pomraning, Kyle R.; Baker, Scott E.

    2015-11-25

    Here we present the draft genome sequence of the dimorphic ascomycete yeast Yarrowia lipolytica strain W29 (ATCC20460TM). Y. lipolytica is a commonly employed model for industrial production of lipases, small molecules, and more recently for its ability to accumulate lipids. It has also been used to study genome evolution in yeast and filamentous fungi due to its position as an early diverging branch of the subphylum Sacchromycotina.

  11. Germinal Center Kinases SmKIN3 and SmKIN24 Are Associated with the Sordaria macrospora Striatin-Interacting Phosphatase and Kinase (STRIPAK) Complex

    PubMed Central

    Frey, Stefan; Reschka, Eva J.; Pöggeler, Stefanie

    2015-01-01

    The striatin-interacting phosphatase and kinase (STRIPAK) complex is composed of striatin, protein phosphatase PP2A and protein kinases that regulate development in animals and fungi. In the filamentous ascomycete Sordaria macrospora, it is required for fruiting-body development and cell fusion. Here, we report on the presence and function of STRIPAK-associated kinases in ascomycetes. Using the mammalian germinal center kinases (GCKs) MST4, STK24, STK25 and MINK1 as query, we identified the two putative homologs SmKIN3 and SmKIN24 in S. macrospora. A BLASTP search revealed that both kinases are conserved among filamentous ascomycetes. The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP). In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution. While deletion of Smkin3 caused larger distances between adjacent septa and increased aerial hyphae, deletion of Smkin24 led to closer spacing of septa and to sterility. Although phenotypically distinct, both kinases appear to function independently because the double-knockout strain ΔSmkin3/ΔSmkin24 displayed the combined phenotypes of each single-deletion strain. PMID:26418262

  12. Estimating the Phanerozoic history of the Ascomycota lineages: combining fossil and molecular data.

    PubMed

    Beimforde, Christina; Feldberg, Kathrin; Nylinder, Stephan; Rikkinen, Jouko; Tuovila, Hanna; Drfelt, Heinrich; Gube, Matthias; Jackson, Daniel J; Reitner, Joachim; Seyfullah, Leyla J; Schmidt, Alexander R

    2014-09-01

    The phylum Ascomycota is by far the largest group in the fungal kingdom. Ecologically important mutualistic associations such as mycorrhizae and lichens have evolved in this group, which are regarded as key innovations that supported the evolution of land plants. Only a few attempts have been made to date the origin of Ascomycota lineages by using molecular clock methods, which is primarily due to the lack of satisfactory fossil calibration data. For this reason we have evaluated all of the oldest available ascomycete fossils from amber (Albian to Miocene) and chert (Devonian and Maastrichtian). The fossils represent five major ascomycete classes (Coniocybomycetes, Dothideomycetes, Eurotiomycetes, Laboulbeniomycetes, and Lecanoromycetes). We have assembled a multi-gene data set (18SrDNA, 28SrDNA, RPB1 and RPB2) from a total of 145 taxa representing most groups of the Ascomycota and utilized fossil calibration points solely from within the ascomycetes to estimate divergence times of Ascomycota lineages with a Bayesian approach. Our results suggest an initial diversification of the Pezizomycotina in the Ordovician, followed by repeated splits of lineages throughout the Phanerozoic, and indicate that this continuous diversification was unaffected by mass extinctions. We suggest that the ecological diversity within each lineage ensured that at least some taxa of each group were able to survive global crises and rapidly recovered. PMID:24792086

  13. Diversity of fungi in creosote-treated crosstie wastes and their resistance to polycyclic aromatic hydrocarbons.

    PubMed

    Kim, Min-Ji; Lee, Hwanhwi; Choi, Yong-Seok; Kim, Gyu-Hyeok; Huh, Na-Yoon; Lee, Sangjoon; Lim, Young Woon; Lee, Sung-Suk; Kim, Jae-Jin

    2010-05-01

    This study was conducted to generate information regarding the diversity of fungi inhabiting creosote-treated wood in a storage yard for crosstie wastes in Gwangmyeong, Korea. Additionally, the resistance to polycyclic aromatic hydrocarbons (PAHs) of indigenous fungi that mainly occupy creosote-treated wood was evaluated. We isolated fungi from the surface and inner area of crosstie wastes and identified them using a combination of traditional methods and molecular techniques. Overall, 179 isolates including 47 different species were isolated from 240 sampling sites. The identified fungal species included 23 ascomycetes, 19 basidiomycetes, and 5 zygomycetes. Three species, Alternaria alternata, Irpex lacteus, and Rhizomucor variabilis, were the most frequently isolated ascomycetes, basidiomycetes, and zygomycetes, respectively. The results of this study showed that there was a large difference in the fungal diversity between the surface and the inner area. Additionally, zygomycetes and ascomycetes were found to have a greater tolerance to PAHs than basidiomycetes. However, two basidiomycetes, Heterobasidion annosum and Schizophyllum commune, showed very high resistance to PAHs, even in response to the highest concentration (1,000 ppm), which indicates that these species may play a role in the degradation of PAHs. PMID:20127413

  14. Evolutionary Divergence of an Elongation Factor 3 from Cryptococcus neoformans

    PubMed Central

    Blakely, Greg; Hekman, James; Chakraburtty, Kalpana; Williamson, Peter R.

    2001-01-01

    Elongation factor 3 (EF3) is considered a promising drug target for the control of fungal diseases because of its requirement for protein synthesis and survival of fungi and a lack of EF3 in the mammalian host. However, EF3 has been characterized only in ascomycete yeast. In order to understand the role of EF3 in a basidiomycete yeast, we cloned the gene encoding EF3 from Cryptococcus neoformans (CnEF3), an important fungal pathogen in immunocompromised patients, including those infected with human immunodeficiency virus. CnEF3 was found to encode a 1,055-amino-acid protein and has 44% identity with EF3 from Saccharomyces cerevisiae (YEF3). Expressed CnEF3 exhibited ATPase activity that was only modestly stimulated by ribosomes from S. cerevisiae. In contrast, CnEF3 showed tight binding to cryptococcal ribosomes, as shown by an inability to be removed under conditions which successfully remove Saccharomyces EF3 from ribosomes (0.5 M KCl or 2 M LiCl). CnEF3 also poorly complemented a YEF3 defect in a diploid null mutant and two temperature-sensitive mutants which have been shown previously to be complemented well by EF3 from other ascomycetes, such as Candida albicans. These data clearly identify the presence of a functioning EF3 in the basidiomycete yeast C. neoformans, which demonstrates an evolutionary divergence from EF3 of ascomycete yeast. PMID:11244063

  15. Soil-inhabiting fungal community composition as qualitative indicator of C metabolism processes

    NASA Astrophysics Data System (ADS)

    Manici, L.; Ciavatta, C.; Caputo, F.

    2009-04-01

    Although fungi represent the greater part of soil microbial biomass, they play an important role in macro-aggregate formation and their carbon (C) assimilation efficiency is markedly higher than that of bacteria (Bailey et al., 2002), they have not yet been widely used as soil biological indicators. The reason is mainly due to the difficulties in application of molecular analysis tools due to limited availability of reference sequence of fungal strains in DNA database and to the low concentration of fungal DNA in soil and in isolating, enumerating and identifying groups of fungi differing for their functioning in soil and for biological characteristics. The presence of Ascomycetes and Basidiomycetes as the two main groups of soil-inhabiting fungi were investigated in four different cropping systems. The soil DNA of soil samples coming from twenty sites (five sites for each system), collected in two cropping systems in northern (soil organic matter - SOM content varying from 0.8 to 1.4 %) and two in southern Italy (SOM from 1.4 to 2.3%), was amplified using Ascomycete-specific primer ITS1F - ITS4A (Larena et al., 1999) and Basidiomycete-specific primer ITS1F -ITS4B (Gardes and Bruns, 1993). On the basis of soil DNA amplified with specific primers, Ascomycetes were much more represented than Basidiomycetes in the cultivated top soil. Basidiomycetes are usually reported to account for more than half of the fungal biomass in undisturbed soils. However the low ratio of Basidiomycete DNA to soil fungal DNA observed in this study could be a feature of soil fungal communities in arable soil affected by desertification problems as those of some Italian cropping systems mainly in Mediterranean area. This phenomenon could be due to soil tillage, which is well known to deeply reduce fungal biomass and to continuous incorporation into the soil of herbaceous crop residues. In fact, Ascomycetes decompose holocellulose in preference to lignin (Oslko & Takeda, 2002) and their growth may depend on readily available energy sources, such as soluble carbohydrates (Hudson, 1968). The high ratio of Ascomycetes in the top layer where crop residues of the recurrent had represented the main substrate for saprophytic fungi could explain these results. On the contrary, Basidiomycetes are the most important synthesizing biomass organisms in forest soils as well as the most effective organisms in lignin decomposition with an important role in humic substances processes (Hurst et al., 1963; Cook and Rayner, 1986). Preliminary results of this study suggest that the composition of soil-inhabiting fungal communities, which are the organisms most involved in C metabolism processes, could represents an useful indicator in programs aimed to increase the quality of organic matter in arable soils. Bailey V., Smith L., Bolton Jr K. 2002. Fungal-to-bacteria ratio investigated for enhanced C sequestration. Soil Biol. Biochem. 34, 997-1007. Cook R., Rayner A.D.M. 1984. Ecology of Saprotrophic Fungi. Longman, London, New York, 415 pp. Gardes M., Bruns T.D. 1993. ITS primers with enhanced specificity for Basidiomycetes: application to the identification of mycorrhizae and rusts. Molec. Ecol. 2, 113-118. Hudson H.J. 1968. The ecology of fungi on plant remains above the soil. New Phytol. 67, 837-874. Hurst H.M., A. Burges, P. Latter. 1963. Some aspects of the biochemistry of humic acid decomposition by fungi. Phytochem. 1, 227-231. Larena I., Salazar O., González V, Julián M.C., Rubio V. 1999. Design of a primer for ribosomal DNA internal transcribed spacer with enhanced specificity for Ascomycetes. J. Biotech. 75, 187-194. Osolko T., Fukasawa Y., Takeda H. 2003. Roles of diverse fungi in Larch neerle-litter decomposition. Mycologia 95, 820-826.

  16. Module evolution and substrate specificity of fungal nonribosomal peptide synthetases involved in siderophore biosynthesis

    PubMed Central

    2008-01-01

    Background Most filamentous ascomycete fungi produce high affinity iron chelators called siderophores, biosynthesized nonribosomally by multimodular adenylating enzymes called nonribosomal peptide synthetases (NRPSs). While genes encoding the majority of NRPSs are intermittently distributed across the fungal kingdom, those encoding ferrichrome synthetase NRPSs, responsible for biosynthesis of ferrichrome siderophores, are conserved, which offers an opportunity to trace their evolution and the genesis of their multimodular domain architecture. Furthermore, since the chemistry of many ferrichromes is known, the biochemical and structural 'rules' guiding NRPS substrate choice can be addressed using protein structural modeling and evolutionary approaches. Results A search of forty-nine complete fungal genome sequences revealed that, with the exception of Schizosaccharomyces pombe, none of the yeast, chytrid, or zygomycete genomes contained a candidate ferrichrome synthetase. In contrast, all filamentous ascomycetes queried contained at least one, while presence and numbers in basidiomycetes varied. Genes encoding ferrichrome synthetases were monophyletic when analyzed with other NRPSs. Phylogenetic analyses provided support for an ancestral duplication event resulting in two main lineages. They also supported the proposed hypothesis that ferrichrome synthetases derive from an ancestral hexamodular gene, likely created by tandem duplication of complete NRPS modules. Recurrent losses of individual domains or complete modules from this ancestral gene best explain the diversity of extant domain architectures observed. Key residues and regions in the adenylation domain pocket involved in substrate choice and for binding the amino and carboxy termini of the substrate were identified. Conclusion Iron-chelating ferrichrome synthetases appear restricted to fission yeast, filamentous ascomycetes, and basidiomycetes and fall into two main lineages. Phylogenetic analyses suggest that loss of domains or modules led to evolution of iterative biosynthetic mechanisms that allow flexibility in biosynthesis of the ferrichrome product. The 10 amino acid NRPS code, proposed earlier, failed when we tried to infer substrate preference. Instead, our analyses point to several regions of the binding pocket important in substrate choice and suggest that two positions of the code are involved in substrate anchoring, not substrate choice. PMID:19055762

  17. The cell end marker Tea4 regulates morphogenesis and pathogenicity in the basidiomycete fungus Ustilago maydis

    PubMed Central

    Valinluck, Michael; Woraratanadharm, Tad; Lu, Ching-yu; Quintanilla, Rene H.

    2014-01-01

    Positional cues localized to distinct cell domains are critical for the generation of cell polarity and cell morphogenesis. These cues lead to assembly of protein complexes that organize the cytoskeleton resulting in delivery of vesicles to sites of polarized growth. Tea4, an SH3 domain protein, was first identified in fission yeast, and is a critical determinant of the axis of polarized growth, a role conserved among ascomycete fungi. Ustilago maydis is a badiomycete fungus that exhibits a yeast-like form that is nonpathogenic and a filamentous form that is pathogenic on maize and teozintle. We are interested in understanding how positional cues contribute to generation and maintenance of these two forms, and their role in pathogenicity. We identified a homologue of fission yeast tea4 in a genetic screen for mutants with altered colony and cell morphology and present here analysis of Tea4 for the first time in a basidiomycete fungus. We demonstrate that Tea4 is an important positional marker for polarized growth and septum location in both forms. We uncover roles for Tea4 in maintenance of cell and neck width, cell separation, and cell wall deposition in the yeast-like form, and in growth rate, formation of retraction septa, growth reversal, and inhibition of budding in the filamentous form. We show that Tea4∷GFP localizes to sites of polarized or potential polarized growth in both forms, as observed in ascomycete fungi. We demonstrate an essential role of Tea4 in pathogencity in the absence of cell fusion. Basidiomycete and ascomycete Tea4 homologues share SH3 and Glc7 domains. Tea4 in basidiomycetes has additional domains, which has led us to hypothesize that Tea4 has novel functions in this group of fungi. PMID:24613993

  18. Evolutionary relationships among putative RNA-dependent RNA polymerases encoded by a mitochondrial virus-like RNA in the Dutch elm disease fungus, Ophiostoma novo-ulmi, by other viruses and virus-like RNAs and by the Arabidopsis mitochondrial genome.

    PubMed

    Hong, Y; Cole, T E; Brasier, C M; Buck, K W

    1998-06-20

    The nucleotide sequence (2617 nucleotides) of virus-like double-stranded (ds) RNA 3a in a diseased isolate, Log1/3-8d2 (Ld), of the ascomycete fungus Ophiostoma novo-ulmi has been determined. One strand of the dsRNA contains an open reading frame (ORF) with the potential to encode a protein of 718 amino acids, and the complementary strand contains two smaller ORFs with the potential to encode proteins of 178 and 182 amino acids, respectively. The large ORF contains 12 UGA codons which code for tryptophan in ascomycete mitochondria and has a codon bias typical of mitochondrial genes, consistent with the localization of Ld dsRNAs within the mitochondria. The amino acid sequence contains motifs characteristic of RNA-dependent RNA polymerases (RdRps). This putative RdRp was shown to be related to putative RdRps of mitochondrial dsRNAs of another ascomycete and a basidiomycete fungus and also to a putative RdRp encoded by the mitochondrial genome of Arabidopsis thaliana. In multiple sequence alignments, the fungal mitochondrial dsRNA-encoded RdRp-like proteins formed a cluster, ancestrally related to the RdRps of the yeast 20S and 23S RNA replicons and of the positive-stranded RNA bacteriophages of the Leviviridae family, but distinct from RdRps of other families and genera of fungal RNA viruses and related plant and animal RNA viruses. Northern blot analysis with RNA 3a strand-specific probes indicated that nucleic acid extracts of Ld contain more single-stranded (positive-stranded) RNA than dsRNA, consistent with an evolutionary relationship between RNA 3a and positive-stranded RNA phages. PMID:9657003

  19. Yeast diversity associated with invasive Dendroctonus valens killing Pinus tabuliformis in China using culturing and molecular methods.

    PubMed

    Lou, Qiao-Zhe; Lu, Min; Sun, Jiang-Hua

    2014-08-01

    Bark beetle-associated yeasts are much less studied than filamentous fungi, yet they are also considered to play important roles in beetle nutrition, detoxification, and chemical communication. The red turpentine beetle, Dendroctonus valens, an invasive bark beetle introduced from North America, became one of the most destructive pests in China, having killed more than 10 million Pinus tabuliformis as well as other pine species. No investigation of yeasts associated with this bark beetle in its invaded ranges has been conducted so far. The aim of this study was to assess the diversity of yeast communities in different microhabitats and during different developmental stages of Den. valens in China using culturing and denaturing gradient gel electrophoresis (DGGE) approaches and to compare the yeast flora between China and the USA. The yeast identity was confirmed by sequencing the D1/D2 domain of LSU ribosomal DNA (rDNA). In total, 21 species (13 ascomycetes and eight basidiomycetes) were detected by culturing method, and 12 species (11 ascomycetes and one basidiomycetes) were detected by molecular methods from China. The most frequent five species in China were Candida piceae (Ogataea clade), Cyberlindnera americana, Candida oregonensis (Metschnikowia clade), Candida nitratophila (Ogataea clade) and an undescribed Saccharomycopsis sp., detected by both methods. Seven species were exclusively detected by DGGE. Ca. oregonensis (Metschnikowia clade) was the most frequently detected species by DGGE method. Eight species (all were ascomycetes) from the USA were isolated; seven of those were also found in China. We found significant differences in yeast total abundance as well as community composition between different developmental stages and significant differences between the surface and the gut. The frass yeast community was more similar to that of Den. valens surface or larvae than to the community of the gut or adults. Possible functions of the yeast associates are discussed. PMID:24691849

  20. Yeasts in malting, with special emphasis on Wickerhamomyces anomalus (synonym Pichia anomala).

    PubMed

    Laitila, Arja; Sarlin, Tuija; Raulio, Mari; Wilhelmson, Annika; Kotaviita, Erja; Huttunen, Timo; Juvonen, Riikka

    2011-01-01

    Malted barley is a major raw material of beer, as well as distilled spirits and several food products. The production of malt (malting) exploits the biochemical reactions of a natural process, grain germination. In addition to germinating grain, the malting process includes another metabolically active component: a diverse microbial community that includes various types of bacteria and fungi. Therefore, malting can be considered as a complex ecosystem involving two metabolically active groups. Yeasts and yeast-like fungi are an important part of this ecosystem, but previously the significance of yeasts in malting has been largely underestimated. Characterization and identification of yeasts in industrial processes revealed 25 ascomycetous yeasts belonging to 10 genera, and 18 basidiomycetous yeasts belonging to 7 genera. In addition, two ascomycetous yeast-like fungi belonging to the genera Aureobasidium and Exophiala were commonly detected. Yeasts and yeast-like fungi produced extracellular hydrolytic enzymes with a potentially positive contribution to the malt enzyme spectrum. Several ascomycetous yeast strains showed strong antagonistic activity against field and storage moulds, Wickerhamomyces anomalus (synonym Pichia anomala) being the most effective species. Malting studies revealed that W. anomalus VTT C-04565 effectively restricted Fusarium growth and hydrophobin production during malting and prevented beer gushing. In order to broaden the antimicrobial spectrum and to improve malt brewhouse performance, W. anomalus could be combined with other starter cultures such as Lactobacillus plantarum. Well-characterized microbial mixtures consisting of barley and malt-derived microbes open up several possibilities to improve malt properties and to ensure the safety of the malting process. PMID:20872177

  1. Yeasts in an industrial malting ecosystem.

    PubMed

    Laitila, A; Wilhelmson, A; Kotaviita, E; Olkku, J; Home, S; Juvonen, R

    2006-11-01

    The malting ecosystem consists of two components: the germinating cereal grains and the complex microbial community. Yeasts and yeast-like fungi are an important part of this ecosystem, but the composition and the effects of this microbial group have been largely unknown. In this study we surveyed the development of yeasts and yeast-like fungi in four industrial scale malting processes. A total of 136 malting process samples were collected and examined for the presence of yeasts growing at 15, 25 and 37 degrees C. More than 700 colonies were isolated and characterized. The isolates were discriminated by PCR-fingerprinting with microsatellite primer (M13). Yeasts representing different fingerprint types were identified by sequence analysis of the D1/D2 domain of the 26S rRNA gene. Furthermore, identified yeasts were screened for the production of alpha-amylase, beta-glucanase, cellulase and xylanase. A numerous and diverse yeast community consisting of both ascomycetous (25) and basidiomycetous (18) species was detected in the various stages of the malting process. The most frequently isolated ascomycetous yeasts belonged to the genera Candida, Clavispora, Galactomyces, Hanseniaspora, Issatchenkia, Pichia, Saccharomyces and Williopsis and the basidiomycetous yeasts to Bulleromyces, Filobasidium, Cryptococcus, Rhodotorula, Sporobolomyces and Trichosporon. In addition, two ascomycetous yeast-like fungi (black yeasts) belonging to the genera Aureobasidium and Exophiala were commonly detected. Yeasts and yeast-like fungi produced extracellular hydrolytic enzymes with a potentially positive contribution to the malt enzyme spectrum. Knowledge of the microbial diversity provides a basis for microflora management and understanding of the role of microbes in the cereal germination process. PMID:16758169

  2. Genomic Mechanisms Accounting for the Adaptation to Parasitism in Nematode-Trapping Fungi

    PubMed Central

    Meerupati, Tejashwari; Andersson, Karl-Magnus; Friman, Eva; Kumar, Dharmendra; Tunlid, Anders; Ahrén, Dag

    2013-01-01

    Orbiliomycetes is one of the earliest diverging branches of the filamentous ascomycetes. The class contains nematode-trapping fungi that form unique infection structures, called traps, to capture and kill free-living nematodes. The traps have evolved differently along several lineages and include adhesive traps (knobs, nets or branches) and constricting rings. We show, by genome sequencing of the knob-forming species Monacrosporium haptotylum and comparison with the net-forming species Arthrobotrys oligospora, that two genomic mechanisms are likely to have been important for the adaptation to parasitism in these fungi. Firstly, the expansion of protein domain families and the large number of species-specific genes indicated that gene duplication followed by functional diversification had a major role in the evolution of the nematode-trapping fungi. Gene expression indicated that many of these genes are important for pathogenicity. Secondly, gene expression of orthologs between the two fungi during infection indicated that differential regulation was an important mechanism for the evolution of parasitism in nematode-trapping fungi. Many of the highly expressed and highly upregulated M. haptotylum transcripts during the early stages of nematode infection were species-specific and encoded small secreted proteins (SSPs) that were affected by repeat-induced point mutations (RIP). An active RIP mechanism was revealed by lack of repeats, dinucleotide bias in repeats and genes, low proportion of recent gene duplicates, and reduction of recent gene family expansions. The high expression and rapid divergence of SSPs indicate a striking similarity in the infection mechanisms of nematode-trapping fungi and plant and insect pathogens from the crown groups of the filamentous ascomycetes (Pezizomycotina). The patterns of gene family expansions in the nematode-trapping fungi were more similar to plant pathogens than to insect and animal pathogens. The observation of RIP activity in the Orbiliomycetes suggested that this mechanism was present early in the evolution of the filamentous ascomycetes. PMID:24244185

  3. Braunomyces dictyosporus gen. sp. nov. from Vietnam.

    PubMed

    Mel'nik, Vadim A; Crous, Pedro W

    2014-06-01

    The generic name Braunomyces (ascomycetes, asexual morph), with B. dictyosporus as type species, is described, illustrated and discussed, based on material collected in Vietnam on leaf debris of an unidentified broadleaved tree. The new genus is well characterised and quite distinct from other synnematous and non-synnematous dematiaceous hyphomycete genera by its unique combination of traits, viz. determinate synnematous conidiomata, integrated, terminal and intercalary conidiogenous cells with one to several conidiogenous vesicles becoming cupulate with age, tretic conidiogenesis, and mostly cruciately septate, solitary dictyoconidia. PMID:25083401

  4. Parallels in amphibian and bat declines from pathogenic fungi.

    PubMed

    Eskew, Evan A; Todd, Brian D

    2013-03-01

    Pathogenic fungi have substantial effects on global biodiversity, and 2 emerging pathogenic species-the chytridiomycete Batrachochytrium dendrobatidis, which causes chytridiomycosis in amphibians, and the ascomycete Geomyces destructans, which causes white-nose syndrome in hibernating bats-are implicated in the widespread decline of their vertebrate hosts. We synthesized current knowledge for chytridiomycosis and white-nose syndrome regarding disease emergence, environmental reservoirs, life history characteristics of the host, and host-pathogen interactions. We found striking similarities between these aspects of chytridiomycosis and white-nose syndrome, and the research that we review and propose should help guide management of future emerging fungal diseases. PMID:23622255

  5. Arxula adeninivorans (Blastobotrys adeninivorans) — A Dimorphic Yeast of Great Biotechnological Potential

    NASA Astrophysics Data System (ADS)

    Böer, Erik; Steinborn, Gerhard; Florschütz, Kristina; Körner, Martina; Gellissen, Gerd; Kunze, Gotthard

    The dimorphic ascomycetous yeast Arxula adeninivorans exhibits some unusual properties. Being a thermo- and halotolerant species it is able to assimilate and ferment many compounds as sole carbon and/or nitrogen source. It utilises n-alkanes and is capable of degrading starch. Due to these unusual biochemical properties A. adeninivorans can be exploited as a gene donor for the production of enzymes with attractive biotechnological characteristics. Examples of A. adeninivorans-derived genes that are overexpressed include the ALIP1 gene encoding a secretory lipase, the AINV encoding invertase, the AXDH encoding xylitol dehydrogenase and the APHY encoding a secretory phosphatase with phytase activity.

  6. Homokaryotic and heterokaryotic hyphae in Terfezia.

    PubMed

    Roth-Bejerano, N; Li, Y-F; Kagan-Zur, V

    2004-02-01

    Mycelia of Terfezia pfeilii (Ascomycetes) were obtained by two methods, i.e., from the sterile hyphae of fresh fruit bodies or by germinating ascospores. Nuclear staining revealed the existence of multinucleate cells in all mycelia. Paired nuclei were observed only in mycelia obtained from sterile hyphae proliferation, while single nuclei were found in mycelia originating from singly germinated spores. Co-cultivation of mycelia from two different ascospores apparently facilitated plasmogamy, resulting in mycelia with paired nuclei. Terfezia boudieri cultures originating from sterile hyphae also exhibit paired nuclei, indicating the possible existence of a long-term heterokaryon. The timing of plasmogamy and karyogamy in Terfezia is discussed. PMID:15031658

  7. Large Scale Sequencing of Dothideomycetes Provides Insights into Genome Evolution and Adaptation

    SciTech Connect

    Haridas, Sajeet; Crous, Pedro; Binder, Manfred; Spatafora, Joseph; Grigoriev, Igor

    2015-03-16

    Dothideomycetes is the largest and most diverse class of ascomycete fungi with 23 orders 110 families, 1300 genera and over 19,000 known species. We present comparative analysis of 70 Dothideomycete genomes including over 50 that we sequenced and are as yet unpublished. This extensive sampling has almost quadrupled the previous study of 18 species and uncovered a 10 fold range of genome sizes. We were able to clarify the phylogenetic positions of several species whose origins were unclear in previous morphological and sequence comparison studies. We analyzed selected gene families including proteases, transporters and small secreted proteins and show that major differences in gene content is influenced by speciation.

  8. Characterization of the BLR1 gene encoding a putative blue-light regulator in the phytopathogenic fungus Bipolaris oryzae.

    PubMed

    Kihara, Junichi; Moriwaki, Akihiro; Tanaka, Nozomi; Ueno, Makoto; Arase, Sakae

    2007-01-01

    Bipolaris oryzae is a filamentous ascomycetous fungus that causes brown leaf spot disease in rice. We isolated and characterized BLR1, a gene that encodes a putative blue-light regulator similar to Neurospora crassa white-collar 1 (WC-1). The deduced amino acid sequence of BLR1 showed high degrees of similarity to other fungal blue-light regulator protein. Disruption of the BLR1 gene demonstrated that this gene is essential for conidial development after conidiophore formation and for near-UV radiation-enhanced photolyase gene expression. PMID:17233721

  9. Molecular data place Trypetheliaceae in Dothideomycetes.

    PubMed

    Del Prado, Ruth; Schmitt, Imke; Kautz, Stefanie; Palice, Zdenek; Lcking, Robert; Lumbsch, H Thorsten

    2006-05-01

    The phylogenetic position of Trypetheliaceae was studied using partial sequences of the mtSSU and nuLSU rDNA of 100 and 110 ascomycetes, respectively, including 48 newly obtained sequences. Our analysis confirms Trypetheliaceae as monophyletic and places the family in Dothideomycetes. Pyrenulaceae, which were previously classified with Trypetheliaceae in Pyrenulales or Melanommatales, are supported as belonging to Chaetothyriomycetes. Monophyly of Pyrenulales, including Trypetheliaceae is rejected using three independent test methods. Monophyly of Arthopyreniaceae plus Trypetheliaceae, the two families including lichen-forming fungi in Dothideomycetes, is also rejected, as well as a placement of Trypetheliaceae in Pleosporales (incl. Melanommatales). PMID:16621495

  10. Invasive Mycotic Infections Caused by Chaetomium perlucidum, a New Agent of Cerebral Phaeohyphomycosis

    PubMed Central

    Barron, M. A.; Sutton, D. A.; Veve, R.; Guarro, J.; Rinaldi, M.; Thompson, E.; Cagnoni, P. J.; Moultney, K.; Madinger, N. E.

    2003-01-01

    We report the first two cases of invasive human mycoses caused by the phaeoid ascomycete, Chaetomium perlucidum, and review the English literature regarding invasive Chaetomium infections. Fatal disseminated disease involving the brain, heart, lungs, and spleen is described in an acute myelogenous leukemia patient. A second patient with a history of asthma and chronic bronchiectasis experiencing right-middle-lobe syndrome grew C. perlucidum from lung tissue. This study adds C. perlucidum to the list of other known neurotropic Chaetomium species, C. atrobrunneum and C. strumarium, and also documents this organism's ability to disseminate beyond the central nervous system. PMID:14605190

  11. [Comparison of the fungal complexes of the Japanese scallop Mizuhopecten yessoensis (Jay, 1856) from different areas of the Peter the Great Bay, Sea of Japan].

    PubMed

    Borzykh, O G; Zvereva, L V

    2014-01-01

    Mycological investigation of the Japanese scallop Mizuhopecten yessoensis (Jay) (Bivalvia) collected in different areas of the Peter the Great Bay, Sea of Japan, was carried out. A total of 72 species of filamentous fungi belonging to 30 genera of ascomycetes, anamorphous fungi, and zygomycetes was isolated from the internal organs of the scallop. The species.diversity of mycelial fungi in the internal organs, especially of members of the generaAspergillus, Penicillium, Cladosporium, and Chaetomium, was found to increase in polluted coastal waters. PMID:25844471

  12. Nucleotide Composition of Nuclear and Mitochondrial Deoxyribonucleic Acid of Fungi

    PubMed Central

    Villa, Vicente D.; Storck, R.

    1968-01-01

    The buoyant density of nuclear and mitochondrial deoxyribonucleic acid (DNA) from 14 species of fungi was determined by CsCl density gradient equilibrium centrifugation. The buoyant density of both types of DNA was the same for all three Mucorales analyzed. The buoyant density of mitochondrial DNA was significantly lower than that of the nuclear DNA for nine species of Ascomycetes and two species of Basidiomycetes. No simple correlation could be obtained from the comparison of the two types of DNA. Mitochondrial DNA represented a very small proportion of total DNA. Heat-denatured mitochondrial DNA renatured more readily than nuclear DNA. PMID:5663568

  13. Fungal diversity on fallen leaves of Ficus in northern Thailand* §

    PubMed Central

    Wang, Hong-kai; Hyde, Kevin D.; Soytong, Kasem; Lin, Fu-cheng

    2008-01-01

    Fallen leaves of Ficus altissima, F. virens, F. benjamina, F. fistulosa and F. semicordata, were collected in Chiang Mai Province in northern Thailand and examined for fungi. Eighty taxa were identified, comprising 56 anamorphic taxa, 23 ascomycetes and 1 basidiomycete. Common fungal species occurring on five host species with high frequency of occurrence were Beltraniella nilgirica, Lasiodiplodia theobromae, Ophioceras leptosporum, Periconia byssoides and Septonema harknessi. Colletotrichum and Stachybotrys were also common genera. The leaves of different Ficus species supported diverse fungal taxa, and the fungal assemblages on the different hosts showed varying overlap. The fungal diversity of saprobes at the host species level is discussed. PMID:18837113

  14. Candida coquimbonensis sp. nov., a link between Australian and Nearctic/Neotropical Phaffomyces.

    PubMed

    Cardinali, Gianluigi; Antonielli, Livio; Corte, Laura; Roscini, Luca; Ganter, Philip F

    2012-12-01

    A novel species of ascomycetous yeast, Candida coquimbonensis sp. nov., from the necrotic tissue of cacti in Chile and Australia is described. C. coquimbonensis sp. nov. is closely related and phenotypically similar to Phaffomyces opuntiae. There is no overlap in the geographical distribution between C. coquimbonensis and any species in the Phaffomyces clade. However, this is the first member of the clade to be collected in both native (Chile) and non-native (Australia) cactus habitats. The type strain of C. coquimbonensis sp. nov. is TSU 00-206.4B(T) ( = CBS 12348(T) = USCFST 12-103(T)). PMID:22729024

  15. Multi-omics analysis reveals regulators of the response to nitrogen limitation in Yarrowia lipolytica

    DOE PAGESBeta

    Pomraning, Kyle R.; Kim, Young -Mo; Nicora, Carrie D.; Chu, Rosalie K.; Bredeweg, Erin L.; Purvine, Samuel O.; Hu, Dehong; Metz, Thomas O.; Baker, Scott E.

    2016-02-25

    Yarrowia lipolytica is an oleaginous ascomycete yeast that stores lipids in response to limitation of nitrogen. Furthermore, while the enzymatic pathways responsible for neutral lipid accumulation in Y. lipolytica are well characterized, regulation of these pathways has received little attention. We therefore sought to characterize the response to nitrogen limitation at system-wide levels, including the proteome, phosphoproteome and metabolome, to better understand how this organism regulates and controls lipid metabolism and to identify targets that may be manipulated to improve lipid yield.

  16. Mildew-Omics: How Global Analyses Aid the Understanding of Life and Evolution of Powdery Mildews

    PubMed Central

    Bindschedler, Laurence V.; Panstruga, Ralph; Spanu, Pietro D.

    2016-01-01

    The common powdery mildew plant diseases are caused by ascomycete fungi of the order Erysiphales. Their characteristic life style as obligate biotrophs renders functional analyses in these species challenging, mainly because of experimental constraints to genetic manipulation. Global large-scale (“-omics”) approaches are thus particularly valuable and insightful for the characterisation of the life and evolution of powdery mildews. Here we review the knowledge obtained so far from genomic, transcriptomic and proteomic studies in these fungi. We consider current limitations and challenges regarding these surveys and provide an outlook on desired future investigations on the basis of the various –omics technologies. PMID:26913042

  17. Metabolites from nematophagous fungi and nematicidal natural products from fungi as alternatives for biological control. Part II: metabolites from nematophagous basidiomycetes and non-nematophagous fungi.

    PubMed

    Degenkolb, Thomas; Vilcinskas, Andreas

    2016-05-01

    In this second section of a two-part mini-review article, we introduce 101 further nematicidal and non-nematicidal secondary metabolites biosynthesized by nematophagous basidiomycetes or non-nematophagous ascomycetes and basidiomycetes. Several of these compounds have promising nematicidal activity and deserve further and more detailed analysis. Thermolides A and B, omphalotins, ophiobolins, bursaphelocides A and B, illinitone A, pseudohalonectrins A and B, dichomitin B, and caryopsomycins A-C are excellent candidates or lead compounds for the development of biocontrol strategies for phytopathogenic nematodes. Paraherquamides, clonostachydiol, and nafuredins offer promising leads for the development of formulations against the intestinal nematodes of ruminants. PMID:26728016

  18. Taming a wild beast: Developing molecular tools and new methods to understand the biology of Zymoseptoria tritici.

    PubMed

    Talbot, Nicholas J

    2015-06-01

    Septoria blotch of wheat is one of the world's most serious plant diseases, which is difficult to control due to the absence of durable host resistance and the increasing frequency of fungicide-resistance. The ascomycete fungus that causes the disease, Zymoseptoria tritici, has been very challenging to study. This special issue of Fungal Genetics and Biology showcases an integrated approach to method development and the innovation of new molecular tools to study the biology of Z. tritici. When considered together, these new methods will have a rapid and dramatic effect on our ability to combat this significant disease. PMID:25975217

  19. Taming a wild beast: Developing molecular tools and new methods to understand the biology of Zymoseptoria tritici

    PubMed Central

    Talbot, Nicholas J.

    2015-01-01

    Septoria blotch of wheat is one of the world’s most serious plant diseases, which is difficult to control due to the absence of durable host resistance and the increasing frequency of fungicide-resistance. The ascomycete fungus that causes the disease, Zymoseptoria tritici, has been very challenging to study. This special issue of Fungal Genetics and Biology showcases an integrated approach to method development and the innovation of new molecular tools to study the biology of Z. tritici. When considered together, these new methods will have a rapid and dramatic effect on our ability to combat this significant disease. PMID:25975217

  20. Larvicidal toxins from Bacillus thuringiensis subspp. kurstaki, morrisoni (strain tenebrionis), and israelensis have no microbicidal or microbiostatic activity against selected bacteria, fungi, and algae in vitro.

    PubMed

    Koskella, J; Stotzky, G

    2002-03-01

    The insecticidal toxins from Bacillus thuringiensis subspp. kurstaki (antilepidopteran), morrisoni strain tenebrionis (anticoleopteran), and israelensis (antidipteran) did not affect the growth of a variety of bacteria (8 gram-negative, 5 gram-positive, and a cyanobacterium), fungi (2 Zygomycetes, 1 Ascomycete, 2 Deuteromycetes, and 2 yeasts), and algae (primarily green and diatoms) in pure and mixed culture, as determined by dilution, disk-diffusion, and sporulation assays with purified free and clay-bound toxins. The insecticidal crystal proteins from B. thuringiensis subspp. kurstaki and israelensis had no antibiotic effect on various gram-positive bacteria. PMID:11989771

  1. Fungal mating pheromones: choreographing the dating game.

    PubMed

    Jones, Stephen K; Bennett, Richard J

    2011-07-01

    Pheromones are ubiquitous from bacteria to mammals - a testament to their importance in regulating inter-cellular communication. In fungal species, they play a critical role in choreographing interactions between mating partners during the program of sexual reproduction. Here, we describe how fungal pheromones are synthesized, their interactions with G protein-coupled receptors, and the signals propagated by this interaction, using Saccharomyces cerevisiae as a reference point. Divergence from this model system is compared amongst the ascomycetes and basidiomycetes, which reveals the wealth of information that has been gleaned from studying pheromone-driven processes across a wide spectrum of the fungal kingdom. PMID:21496492

  2. The diversity of yeasts associated with grapes and musts of the Strekov winegrowing region, Slovakia.

    PubMed

    Nemcová, Kornélia; Breierová, Emília; Vadkertiová, Renáta; Molnárová, Jana

    2015-03-01

    Many different yeast species have been isolated from grapes and musts worldwide. The diversity and frequency of yeasts depend on a number of factors such as the grape variety, the physical damage of the grapes, the weather conditions and the chemical composition of must. A total of 366 isolates were associated with the three grape cultivars: Blue Frankish, Green Veltliner and Sauvignon blanc over four consecutive years. Yeast cultures were isolated from the grapes and from the fermenting musts after the first and seventh days. The ascomycetous yeasts of the genera Aureobasidium, Candida, Hanseniaspora, Metschnikowia, Pichia, Saccharomyces and Saccharomycopsis together with basidiomycetous yeasts of the genera Cryptococcus, Dioszegia, Filobasidium, Rhodotorula and Sporidiobolus were associated with the three grape varieties. Hanseniaspora uvarum, Metschnikowia pulcherrima, Pichia kluyveri, Pichia kudriavzevii and Sporidiobolus pararoseus were found on the berries in significant amounts. P. kluyveri and P. kudriavzevii were more associated with the damaged grapes, whereas Sp. pararoseus with intact ones. H. uvarum and M. pulcherrima were present on both types of grapes almost equally. The yeast composition and quantitative representation of yeast species varied over the grape varieties and the years examined. Although the basidiomycetous species formed a significant proportion of the yeast population in some individual grape variety/year combinations, the ascomycetous species were dominant. PMID:25253264

  3. Zygomycetes in Vesicular Basanites from Vesteris Seamount, Greenland Basin--A New Type of Cryptoendolithic Fungi.

    PubMed

    Ivarsson, Magnus; Peckmann, Jörn; Tehler, Anders; Broman, Curt; Bach, Wolfgang; Behrens, Katharina; Reitner, Joachim; Böttcher, Michael E; Norbäck Ivarsson, Lena

    2015-01-01

    Fungi have been recognized as a frequent colonizer of subseafloor basalt but a substantial understanding of their abundance, diversity and ecological role in this environment is still lacking. Here we report fossilized cryptoendolithic fungal communities represented by mainly Zygomycetes and minor Ascomycetes in vesicles of dredged volcanic rocks (basanites) from the Vesteris Seamount in the Greenland Basin. Zygomycetes had not been reported from subseafloor basalt previously. Different stages in zygospore formation are documented in the studied samples, representing a reproduction cycle. Spore structures of both Zygomycetes and Ascomycetes are mineralized by romanechite-like Mn oxide phases, indicating an involvement in Mn(II) oxidation to form Mn(III,VI) oxides. Zygospores still exhibit a core of carbonaceous matter due to their resistance to degradation. The fungi are closely associated with fossiliferous marine sediments that have been introduced into the vesicles. At the contact to sediment infillings, fungi produced haustoria that penetrated and scavenged on the remains of fragmented marine organisms. It is most likely that such marine debris is the main carbon source for fungi in shallow volcanic rocks, which favored the establishment of vital colonies. PMID:26181773

  4. Identification of fungal cell wall mutants using susceptibility assays based on Calcofluor white and Congo red.

    PubMed

    Ram, Arthur F J; Klis, Frans M

    2006-01-01

    The fungal cell wall is an essential organelle and represents a considerable metabolic investment. Its macromolecular composition, molecular organization and thickness can vary greatly depending on environmental conditions. Its construction is also tightly controlled in space and time. Many genes are therefore involved in building the fungal cell wall. Here we present a simple approach for detecting these genes. The method is based on the observation that cell wall mutants are generally more sensitive to two related anionic dyes, Calcofluor white (CFW) and Congo red (CR), both of which interfere with the construction and stress response of the cell wall. CFW-based and CR-based susceptibility assays identify cell wall mutants not only in ascomycetous yeasts (such as Saccharomyces cerevisiae and Candida albicans) but also in mycelial ascomycetes (such as Aspergillus fumigatus and Aspergillus niger), basidiomycetous species (Cryptococcus neoformans) and probably also zygomycetous fungi. The protocol can be completed in 4-6 h (excluding the incubation time required for fungal growth). PMID:17406464

  5. Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis.

    PubMed

    Oliveira, Manuela; Amorim, M Isabel; Ferreira, Elsa; Delgado, Luís; Abreu, Ilda

    2010-04-01

    The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and beta-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or beta-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores. PMID:20143229

  6. Yeast communities in Sphagnum phyllosphere along the temperature-moisture ecocline in the boreal forest-swamp ecosystem and description of Candida sphagnicola sp. nov.

    PubMed

    Kachalkin, Aleksey V; Yurkov, Andrey M

    2012-06-01

    The effects of the temperature-moisture factors on the phylloplane yeast communities inhabiting Sphagnum mosses were studied along the transition from a boreal forest to a swamp biotope at the Central Forest State Biosphere Reserve (Tver region, Russia). We tested the hypothesis that microclimatic parameters affect yeast community composition and structure even on a rather small spatial scale. Using a conventional plating technique we isolated and identified by molecular methods a total of 15 species of yeasts. Total yeast counts and species richness values did not depend on environmental factors, although yeast community composition and structure did. On average, Sphagnum in the swamp biotope supported a more evenly structured yeast community. Relative abundance of ascomycetous yeasts was significantly higher on swamp moss. Rhodotorula mucilaginosa dominated in the spruce forest and Cryptococcus magnus was more abundant in the swamp. Our study confirmed the low occurrence of tremellaceous yeasts in the Sphagnum phyllosphere. Of the few isolated ascomycetous yeast and yeast-like species, some were differentiated from hitherto known species in physiological tests and phylogenetic analyses. We describe one of them as Candida sphagnicola and designate KBP Y-3887(T) (=CBS 11774(T) = VKPM Y-3566(T) = MUCL 53590(T)) as the type strain. The new species was registered in MycoBank under MB 563443. PMID:22331450

  7. Complex evolution of the DAL5 transporter family

    PubMed Central

    Hellborg, Linda; Woolfit, Megan; Arthursson-Hellborg, Mattias; Piškur, Jure

    2008-01-01

    Background Genes continuously duplicate and the duplicated copies remain in the genome or get deleted. The DAL5 subfamily of transmembrane transporter genes has eight known members in S. cerevisiae. All are putative anion:cation symporters of vitamins (such as allantoate, nicotinate, panthotenate and biotin). The DAL5 subfamily is an old and important group since it is represented in both Basidiomycetes ("mushrooms") and Ascomycetes ("yeast"). We studied the complex evolution of this group in species from the kingdom of fungi particularly the Ascomycetes. Results We identified numerous gene duplications creating sets of orthologous and paralogous genes. In different lineages the DAL5 subfamily members expanded or contracted and in some lineages a specific member could not be found at all. We also observed a close relationship between the gene YIL166C and its homologs in the Saccharomyces sensu stricto species and two "wine spoiler" yeasts, Dekkera bruxellensis and Candida guilliermondi, which could possibly be the result of horizontal gene transfer between these distantly related species. In the analyses we detect several well defined groups without S. cerevisiae representation suggesting new gene members in this subfamily with perhaps altered specialization or function. Conclusion The transmembrane DAL5 subfamily was found to have a very complex evolution in yeast with intra- and interspecific duplications and unusual relationships indicating specialization, specific deletions and maybe even horizontal gene transfer. We believe that this group will be important in future investigations of evolution in fungi and especially the evolution of transmembrane proteins and their specialization. PMID:18405355

  8. The conserved global regulator VeA is necessary for symptom production and mycotoxin synthesis in maize seedlings by Fusarium verticillioides

    PubMed Central

    Myung, K.; Zitomer, N. C.; Duvall, M.; Glenn, A. E.; Riley, R. T.; Calvo, A. M.

    2011-01-01

    The veA or velvet gene is necessary for biosynthesis of mycotoxins and other secondary metabolites in Aspergillus species. In addition, veA has also been demonstrated to be necessary for normal seed colonization in Aspergillus flavus and Aspergillus parasiticus. The present study shows that veA homologues are broadly distributed in fungi, particularly in Ascomycetes. The Fusarium verticillioides veA orthologue, FvVE1, is also required for the synthesis of several secondary metabolites, including fumonisin and fusarins. This study also shows that maize plants grown from seeds inoculated with FvVE1 deletion mutants did not show disease symptoms, while plants grown from seeds inoculated with the F. verticillioides wildtype and complementation strains clearly showed disease symptoms under the same experimental conditions. In this latter case, the presence of lesions coincided with accumulation of fumonisins in the plant tissues, and only these plant tissues had elevated levels of sphingoid bases and their 1-phosphate derivatives, indicating inhibition of ceramide synthase and disruption of sphingolipid metabolism. The results strongly suggest that FvVE1 is necessary for pathogenicity by F. verticillioides against maize seedlings. The conservation of veA homologues among ascomycetes suggests that veA could play a pivotal role in regulating secondary metabolism and associated pathogenicity in other fungi. PMID:22247572

  9. Phenotype of a Mechanosensitive Channel Mutant, mid-1, in a Filamentous Fungus, Neurospora crassa▿

    PubMed Central

    Lew, Roger R.; Abbas, Zohaib; Anderca, Marinela I.; Free, Stephen J.

    2008-01-01

    In the yeast Saccharomyces cerevisiae, the MID1 (mating-induced death) gene encodes a stretch-activated channel which is required for successful mating; the mutant phenotype is rescued by elevated extracellular calcium. Homologs of the MID1 gene are found in fungi that are morphologically complex compared to yeast, both Basidiomycetes and Ascomycetes. We explored the phenotype of a mid-1 knockout mutant in the filamentous ascomycete Neurospora crassa. The mutant exhibits lower growth vigor than the wild type (which is not rescued by replete calcium) and mates successfully. Thus, the role of the MID-1 protein differs from that of the homologous gene product in yeast. Hyphal cytology, growth on diverse carbon sources, turgor regulation, and circadian rhythms of the mid-1 mutant are all similar to those of the wild type. However, basal turgor is lower than wild type, as is the activity of the plasma membrane H+-ATPase (measured by cyanide [CN−]-induced depolarization of the energy-dependent component of the membrane potential). In addition, the mutant is unable to grow at low extracellular Ca2+ levels or when cytoplasmic Ca2+ is elevated with the Ca2+ ionophore A23187. We conclude that the MID-1 protein plays a role in regulation of ion transport via Ca2+ homeostasis and signaling. In the absence of normal ion transport activity, the mutant exhibits poorer growth. PMID:18296620

  10. [Lipid Composition in Cell Walls and in Mycelial and Spore Cells of Mycelial Fungi].

    PubMed

    Feofilova, E P; Sergeeva, Ya E; Mysyakina, I S; Bokareva, D A

    2015-01-01

    Qualitative and quantitative differences were found between the lipids of cell walls (CW) and of whole mycelial cells and dormant cells of mucoraceous and ascomycete fungi. Thus, whole mycelial cells (WC) contained more lipids than CW. Unlike sporangiospores and conidia (exogenous dormant spores), zygotes were found to have the highest content of triacylglycerol lipids (70%). Cell walls of mucoraceous fungi contained more triacylglycerols (TAG) and less polar lipids than ascomycete lipids. While all CW and WC studied were similar in fatty acid (FA) composition, their ratio was specific for each structure: linoleic acid predominated in mycelial CW and WC, while oleic acid was predominant in the spores; this difference was especially pronounced in conidial WC. Unlike WC, in CW massive lipids may be represented not by phosphatidylethanolamine (PEA) and phosphatidylcholine (PC), but by free fatty acids (FFA), free (FSt) and etherified sterols (ESt), phosphatidic acid (PA), fatty acid methyl esters (FAME), and glycolipids (GL), which is an indication of a special functional role of CW. PMID:26263626

  11. New, rare or remarkable microfungi in the Italian Alps (Carnic Alps)--part I--ascomycotina.

    PubMed

    Feige, G B; Ale-Agha, N; Jensen, M; Christiaans, B; Kricke, R

    2004-01-01

    During our observations in the SE part of the Carnic Alps in the year 2003 we were able to collect and identify 35 ascomycetes on trees and dead wood. Among these one can find numerous ascomycetes of different orders e.g. Pyrenomycetes, Loculoascomycetes and Discomycetes. Some species like Botryosphaeria ribis GROSENLUCHER & DUGGAR on Ribes alpinum L., Dothiora pyrenophora (FR.) FR. on Sorbus aucuparia L., Gemmamyces piceae (BORTH.) CASAGO. on Picea excelsa (LAM.) LINK, Glomerella montana (SACC.) v. ARX & E. MULLER on Sesleria caerulea (L.) ARD, Hymenoscyphus immutabilis (Fuck.) Dennis on Alnus incana (L.) Moench, Hysterographium fraxini (PERS. Ex. FR.) de Not. on Fraxinus ornus L., Lachnellula willkommii (Hartig) DENNIS [= Trichascyphella willkommii (Hartig) NANNF.] on Larix decidua MILL.,Leptosphaeria lycopodina (Mont.) SACC. on Lycopodium annotinum L., Mollisia adenostylidis REHM. on Adenostyles glabra (MILL.) DC., Pezicula cinnamomea (DC.)SACC. [ana: Cryptosporiopsis quercina PETRAK] on Quercus robur L., Pyrenopeziza petiolaris (A. & S. Ex FR.) NANNF. on Acer pseudoplatanus L., Tapesia rosae (PERS.) FUCKEL on Rosa canina L., are new for this area. All specimen are deposited in the Herbarium ESS Mycotheca Parva, Collection G.B. Feige/N. Ale-Agha. PMID:15756826

  12. Diverse ecological roles within fungal communities in decomposing logs of Picea abies.

    PubMed

    Ottosson, Elisabet; Kubartová, Ariana; Edman, Mattias; Jönsson, Mari; Lindhe, Anders; Stenlid, Jan; Dahlberg, Anders

    2015-03-01

    Fungal communities in Norway spruce (Picea abies) logs in two forests in Sweden were investigated by 454-sequence analyses and by examining the ecological roles of the detected taxa. We also investigated the relationship between fruit bodies and mycelia in wood and whether community assembly was affected by how the dead wood was formed. Fungal communities were highly variable in terms of phylogenetic composition and ecological roles: 1910 fungal operational taxonomic units (OTUs) were detected; 21% were identified to species level. In total, 58% of the OTUs were ascomycetes and 31% basidiomycetes. Of the 231 337 reads, 38% were ascomycetes and 60% basidiomycetes. Ecological roles were assigned to 35% of the OTUs, accounting for 62% of the reads. Wood-decaying fungi were the most common group; however, other saprotrophic, mycorrhizal, lichenized, parasitic and endophytic fungi were also common. Fungal communities in logs formed by stem breakage were different to those in logs originating from butt breakage or uprooting. DNA of specific species was detected in logs many years after the last recorded fungal fruiting. Combining taxonomic identification with knowledge of ecological roles may provide valuable insights into properties of fungal communities; however, precise ecological information about many fungal species is still lacking. PMID:25764460

  13. Addressing Inter-Gene Heterogeneity in Maximum Likelihood Phylogenomic Analysis: Yeasts Revisited

    PubMed Central

    Hess, Jaqueline; Goldman, Nick

    2011-01-01

    Phylogenomic approaches to the resolution of inter-species relationships have become well established in recent years. Often these involve concatenation of many orthologous genes found in the respective genomes followed by analysis using standard phylogenetic models. Genome-scale data promise increased resolution by minimising sampling error, yet are associated with well-known but often inappropriately addressed caveats arising through data heterogeneity and model violation. These can lead to the reconstruction of highly-supported but incorrect topologies. With the aim of obtaining a species tree for 18 species within the ascomycetous yeasts, we have investigated the use of appropriate evolutionary models to address inter-gene heterogeneities and the scalability and validity of supermatrix analysis as the phylogenetic problem becomes more difficult and the number of genes analysed approaches truly phylogenomic dimensions. We have extended a widely-known early phylogenomic study of yeasts by adding additional species to increase diversity and augmenting the number of genes under analysis. We have investigated sophisticated maximum likelihood analyses, considering not only a concatenated version of the data but also partitioned models where each gene constitutes a partition and parameters are free to vary between the different partitions (thereby accounting for variation in the evolutionary processes at different loci). We find considerable increases in likelihood using these complex models, arguing for the need for appropriate models when analyzing phylogenomic data. Using these methods, we were able to reconstruct a well-supported tree for 18 ascomycetous yeasts spanning about 250 million years of evolution. PMID:21850235

  14. Relationships among genera of the Saccharomycotina (Ascomycota) from multigene phylogenetic analysis of type species.

    PubMed

    Kurtzman, Cletus P; Robnett, Christie J

    2013-02-01

    Relationships among ascomycetous yeast genera (subphylum Saccharomycotina, phylum Ascomycota) have been uncertain. In the present study, type species of 70 currently recognized genera are compared from divergence in the nearly entire nuclear gene sequences for large subunit rRNA, small subunit (SSU) rRNA, translation elongation factor-1α, and RNA polymerase II, subunits 1 (RPB1) and 2 (RPB2). The analysis substantiates earlier proposals that all known ascomycetous yeast genera now assigned to the Saccharomycotina represent a single clade. Maximum likelihood analysis resolved the taxa into eight large multigenus clades and four-one- and two-genus clades. Maximum parsimony and neighbor-joining analyses gave similar results. Genera of the family Saccharomycetaceae remain as one large clade as previously demonstrated, to which the genus Cyniclomyces is now assigned. Pichia, Saturnispora, Kregervanrija, Dekkera, Ogataea and Ambrosiozyma are members of a single large clade, which is separate from the clade that includes Barnettozyma, Cyberlindnera, Phaffomyces, Starmera and Wickerhamomyces. Other clades include Kodamaea, Metschnikowia, Debaryomyces, Cephaloascus and related genera, which are separate from the clade that includes Zygoascus, Trichomonascus, Yarrowia and others. This study once again demonstrates that there is limited congruence between a system of classification based on phenotype and a system determined from DNA sequences. PMID:22978764

  15. Purification, characterization, and amino acid sequence of cerato-platanin, a new phytotoxic protein from Ceratocystis fimbriata f. sp. platani.

    PubMed

    Pazzagli, L; Cappugi, G; Manao, G; Camici, G; Santini, A; Scala, A

    1999-08-27

    A new phytotoxic protein (cerato-platanin) of about 12.4 kDa has been identified in culture filtrates of the Ascomycete Ceratocystis fimbriata f. sp. platani, the causal agent of canker stain disease. The toxicity of the pure protein was bioassayed by detecting the inducing necrosis in tobacco leaves. The pure protein also elicited host synthesis of fluorescent substances in tobacco and plane (Platanus acerifolia) leaves. We purified the protein from culture medium to homogeneity. Its complete amino acid sequence was determined; this protein consists of 120 amino acid residues, contains 4 cysteines (S-S-bridged), and has a high percentage of hydrophobic residues. The molecular weight calculated from the amino acid sequence agrees with that determined by mass spectrometry, suggesting that no post-transnational modification occurs. Searches performed by the BLAST program in data banks (Swiss-Prot, EBI, and GenBank(TM)) revealed that this protein is highly homologous with two proteins produced by other Ascomycete fungi. One, produced during infection of wheat leaves, is codified by the snodprot1 gene of Phaeosphaeria nodorum (the causal agent of glume blotch of wheat), whereas the other is the rAsp f13 allergen from Aspergillus fumigatus. Furthermore, the N terminus of cerato-platanin is homologous with that of cerato-ulmin, a phytotoxic protein belonging to the hydrophobin family and produced by Ophiostoma (Ceratocystis) ulmi, a fungus responsible for Dutch elm disease. PMID:10455173

  16. Cloning and sequence analysis of the MAT-B (MAT-2) genes from the three Dutch elm disease pathogens, Ophiostoma ulmi, O. novo-ulmi, and O. himal-ulmi.

    PubMed

    Paoletti, Mathieu; Buck, Kenneth W; Brasier, Clive M

    2005-09-01

    There were two successive pandemics of Dutch Elm Disease (DED) in Europe, parts of Asia and North America in the last century, caused by two ascomycete fungal species, Ophiostoma ulmi and O. novo-ulmi. A third DED species, O. himal-ulmi, was later discovered in the Himalayas. For each of these three species, we now report on the cloning and analysis of a 2.2 kb sequence containing the coding region and 5' and 3' flanking sequences of the mating type B (MAT-B) gene, which is involved in the control of sexual compatibility. The amino acid sequence of the single protein encoded by the gene for each species contained a conserved DNA-binding motif called the high mobility group (HMG) box which showed significant sequence similarity to corresponding sequences in many ascomycete MAT-2 genes. Phylogenetic trees constructed from the MAT-B (renamed MAT-2) nucleotide and derived amino acid sequences showed distinct clades corresponding to the three Ophiostoma species and a clear separation of the O. novo-ulmi clade into the two subspecies americana and novo-ulmi. The 3' flanking regions have been shown to contain variable numbers of repeated oligonucleotide sequences, the number of which is species-specific and readily distinguished by a simple PCR assay. PMID:16209304

  17. Mycological and ecotoxicological characterisation of landfill leachate before and after traditional treatments.

    PubMed

    Tigini, Valeria; Prigione, Valeria; Varese, Giovanna Cristina

    2014-07-15

    Pollution caused by landfill leachates is one of the main problems of urbanised areas, on account of their chemical composition, which turn in an ineffective treatment. A characterisation of leachates, which takes into account chemical, ecotoxicological and mycological aspects, is basilar for the evaluation of environmental impact of leachate and the development of suitable treatment techniques. In this study, the toxicity of a raw leachate and an effluent coming from traditional wastewater treatment plant was assessed by means of 4 ecotoxicological assays. Both the samples exceed the legal threshold value according to all the tested organisms, indicating the ineffectiveness of activated sludge treatment in the reduction of toxicity. The autochthonous mycoflora of the two samples was evaluated by filtration. The fungal load was 73 CFU for leachate and 102 CFU for the effluent. Ascomycetes were the dominant fraction (81% and 61%, for leachate and effluent respectively), followed by basidiomycetes (19% and 39%, respectively). Most of them were potential emerging pathogens. A decolourisation screening with autochthonous fungi was set up towards both samples in the presence or absence of glucose. Eleven fungi (basidiomycetes and ascomycetes) achieved up to 38% decolourisation yields, showing to be promising fungi for the bioremediation of leachates. Further experiment will be aimed to the study of decolourisation mechanism and toxicity reduction. PMID:24793330

  18. Implications of cellobiohydrolase glycosylation for use in biomass conversion.

    PubMed

    Jeoh, Tina; Michener, William; Himmel, Michael E; Decker, Stephen R; Adney, William S

    2008-01-01

    The cellulase producing ascomycete, Trichoderma reesei (Hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. It is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. During the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. Another ascomycete, Aspergillus niger var. awamori, is also known to produce large quantities of heterologous proteins for industry. In this study, T. reesei Cel7A, a cellobiohydrolase, was expressed in A. niger var. awamori and subjected to detailed biophysical characterization. The purified recombinant enzyme contains six times the amount of N-linked glycan than the enzyme purified from a commercial T. reesei enzyme preparation. The activities of the two enzyme forms were compared using bacterial (microcrystalline) and phosphoric acid swollen (amorphous) cellulose as substrates. This comparison suggested that the increased level of N-glycosylation of the recombinant Cel7A (rCel7A) resulted in reduced activity and increased non-productive binding on cellulose. When treated with the N-glycosidase PNGaseF, the molecular weight of the recombinant enzyme approached that of the commercial enzyme and the activity on cellulose was improved. PMID:18471276

  19. Performance of four ribosomal DNA regions to infer higher-level phylogenetic relationships of inoperculate euascomycetes (Leotiomyceta).

    PubMed

    Lumbsch, H Thorsten; Schmitt, Imke; Lindemuth, Ralf; Miller, Andrew; Mangold, Armin; Fernandez, Fernando; Huhndorf, Sabine

    2005-03-01

    The inoperculate euascomycetes are filamentous fungi that form saprobic, parasitic, and symbiotic associations with a wide variety of animals, plants, cyanobacteria, and other fungi. The higher-level relationships of this economically important group have been unsettled for over 100 years. A data set of 55 species was assembled including sequence data from nuclear and mitochondrial small and large subunit rDNAs for each taxon; 83 new sequences were obtained for this study. Parsimony and Bayesian analyses were performed using the four-region data set and all 14 possible subpartitions of the data. The mitochondrial LSU rDNA was used for the first time in a higher-level phylogenetic study of ascomycetes and its use in concatenated analyses is supported. The classes that were recognized in Leotiomyceta (=inoperculate euascomycetes) in a classification by Eriksson and Winka [Myconet 1 (1997) 1] are strongly supported as monophyletic. The following classes formed strongly supported sister-groups: Arthoniomycetes and Dothideomycetes, Chaetothyriomycetes and Eurotiomycetes, and Leotiomycetes and Sordariomycetes. Nevertheless, the backbone of the euascomycete phylogeny remains poorly resolved. Bayesian posterior probabilities were always higher than maximum parsimony bootstrap values, but converged with an increase in gene partitions analyzed in concatenated analyses. Comparison of five recent higher-level phylogenetic studies in ascomycetes demonstrates a high degree of uncertainty in the relationships between classes. PMID:15683926

  20. Whatever happened to the pyrenomycetes and loculoascomycetes?

    PubMed

    Lumbsch, H Thorsten; Huhndorf, Sabine M

    2007-09-01

    An overview of current phylogenetic studies employing molecular data to test previously formulated hypotheses of relationships of loculoascomycetes and pyrenomycetes is given, concentrating on three topics: (1) circumscription and classification of loculoascomycetes, (2) a new classification of Sordariales, and (3) the phylogenetic occurrence of lichenized pyrenomycetes. With regard to these three examples, our review indicates: (1) In traditional taxonomy ascomycetes were classified according to their ascoma-types, with the class Pyrenomycetes including all taxa having perithecia. Later, the development of ascomata and the type of ascus were employed for higher-level classification, and consequently, Loculoascomycetes was separated from Pyrenomycetes. However, molecular studies show that even these revised classifications were too coarse. The Loculoascomycetes fall into two distinct and not closely related groups, which are placed in two clades: Chaetothyriomycetidae and Dothideomycetes. (2) Ascospore morphology has been widely used in taxonomy of ascomycetes, and Sordariales is a prominent example of this. Molecular data suggest that ascomatal wall morphology is a better predictor of phylogenetic relationships in these fungi. Further, the molecular data helped to redefine the circumscription of Sordariales. (3) The majority of non-lichenized pyrenomycetes form a monophyletic group: Sordariomycetes. However, the lichenized pyrenomycetes are highly polyphyletic. Pyrenocarpous lichen-forming fungi occur in several lineages each in Dothideomycetes, Chaetothyriomycetidae, and Lecanoromycetes, whereas no lichenized forms are currently known in the classical pyrenomycetous Sordariomycetes. PMID:18029164

  1. The obligately lichenicolous genus Lichenoconium represents a novel lineage in the Dothideomycetes.

    PubMed

    Lawrey, James D; Diederich, Paul; Nelsen, Matthew P; Sikaroodi, Masoumeh; Gillevet, Patrick M; Brand, A Maarten; van den Boom, Pieter

    2011-02-01

    Lichenicolous fungi are obligately lichen-associated organisms that have evolved many times throughout the Ascomycota and Basidiomycota. Approximately 20% of lichenicolous ascomycetes are recognized only from asexual (anamorphic) characteristics, so the phylogenetic position of many groups has never been resolved. Here we present the first molecular phylogeny of Lichenoconium, a genus of strictly asexual, obligately lichenicolous species with broad geographic distributions and diverse host ecologies. We obtained nuclear and mitochondrial rDNA sequences from fungal cultures isolated from four species in the genus, including a new species, Lichenoconium aeruginosum sp. nov., collected in France, Luxembourg and Netherlands. Our multilocus phylogeny supports the monophyly of fungi in the genus Lichenoconium, and places the genus in the Dothideomycetes, an ascomycete class made up mainly of saprobes and plant-associated endophytes and pathogens. There are only a few recognized groups of lichen-formers in the Dothideomycetes, but Lichenoconium is not supported as being closely related to any of these, nor to any other recognized order within the Dothideomycetes. Given that Lichenoconium is but one of over 100 genera of anamorphic lichenicolous fungi, most of which have never been studied phylogenetically, we suggest that asexual lichenicolous fungi may represent novel and evolutionarily significant phylogenetic groups in the Kingdom Fungi. PMID:21315315

  2. Ribosomal and RPB2 DNA sequence analyses suggest that Sporidesmium and morphologically similar genera are polyphyletic.

    PubMed

    Shenoy, Belle Damodara; Jeewon, Rajesh; Wu, Wenping P; Bhat, Darbhe Jayarama; Hyde, Kevin D

    2006-08-01

    Sporidesmium and morphologically similar dematiaceous, hyphomycetous genera are characterised by holoblastic phragmoconidia produced on proliferating or non-proliferating conidiophores. They include a number of asexual (anamorphic) genera taxonomically segregated from Sporidesmium sensu lato and are similar in having schizolytic conidial secession. The taxonomy of these ubiquitous asexual fungi and their affinities with known Ascomycetes are, however, still obscure. This study incorporates a phylogenetic investigation, based on the LSU nu-rDNA and RNA polymerase II second largest subunit (RPB2) gene sequence, to assess the possible familial placement of Ellisembia, Linkosia, Repetophragma, Sporidesmiella, Sporidesmium and Stanjehughesia, and justify whether anamorphic characters are proper phylogenetic indicators. Phylogenies provide conclusive evidence to suggest that Sporidesmium is not monophyletic and species are phylogenetically distributed in two major ascomycete classes, Dothideomycetes and Sordariomycetes. Morphologies currently used in their classification have undergone convergent evolution and are not phylogenetically reliable. The possible teleomorphic affinities of these anamorphic genera are discussed in light of morphology and molecular data. As these anamorphs, in most cases, are the sole known morph of the holomorph, it is proposed that in the absence of or failure to detect their teleomorphic phase, the anamorph names should be used for the holomorph. PMID:16908125

  3. Community composition of root-associated fungi in a Quercus-dominated temperate forest: "codominance" of mycorrhizal and root-endophytic fungi.

    PubMed

    Toju, Hirokazu; Yamamoto, Satoshi; Sato, Hirotoshi; Tanabe, Akifumi S; Gilbert, Gregory S; Kadowaki, Kohmei

    2013-05-01

    In terrestrial ecosystems, plant roots are colonized by various clades of mycorrhizal and endophytic fungi. Focused on the root systems of an oak-dominated temperate forest in Japan, we used 454 pyrosequencing to explore how phylogenetically diverse fungi constitute an ecological community of multiple ecotypes. In total, 345 operational taxonomic units (OTUs) of fungi were found from 159 terminal-root samples from 12 plant species occurring in the forest. Due to the dominance of an oak species (Quercus serrata), diverse ectomycorrhizal clades such as Russula, Lactarius, Cortinarius, Tomentella, Amanita, Boletus, and Cenococcum were observed. Unexpectedly, the root-associated fungal community was dominated by root-endophytic ascomycetes in Helotiales, Chaetothyriales, and Rhytismatales. Overall, 55.3% of root samples were colonized by both the commonly observed ascomycetes and ectomycorrhizal fungi; 75.0% of the root samples of the dominant Q. serrata were so cocolonized. Overall, this study revealed that root-associated fungal communities of oak-dominated temperate forests were dominated not only by ectomycorrhizal fungi but also by diverse root endophytes and that potential ecological interactions between the two ecotypes may be important to understand the complex assembly processes of belowground fungal communities. PMID:23762515

  4. A class III histidine kinase acts as a novel virulence factor in Botrytis cinerea.

    PubMed

    Viaud, Muriel; Fillinger, Sabine; Liu, Weiwei; Polepalli, Jai Santosh; Le Pêcheur, Pascal; Kunduru, Aditya Reddy; Leroux, Pierre; Legendre, Laurent

    2006-09-01

    Filamentous ascomycetes contain large numbers of histidine kinases (HK) that belong to eleven classes. Members of class III from different species were previously shown to be involved in osmoregulation and resistance to dicarboximide and phenylpyrrole fungicides. We have inactivated the gene encoding the single group III HK, BOS1, in the economically important plant pathogen Botrytis cinerea. BOS1 inactivation had pleiotropic effects on the fungus. Besides the expected osmosensitivity and resistance to fungicides, null mutants presented additional characteristics indicating that BOS1 is necessary for normal macroconidiation and full virulence. On standard culture media, null mutants very rarely formed conidiophores and those few conidiophores failed to produce conidia. This defect could be partially restored with 1 M sorbitol, suggesting that another BOS1-independent signal cascade may be involved in macroconidiation. The mutants were not found to be hypersensitive to various oxidative stresses but were more resistant to menadione. Finally, pathogenicity tests showed that bos1-null mutants were significantly reduced in the ability to infect host plants. Appressorium morphogenesis was not altered; however, in planta growth was severely reduced. To our knowledge, this is the first class III HK characterized as a pathogenicity factor in a plant-pathogenic ascomycete. PMID:16941908

  5. Phylogeny and origin of 82 zygomycetes from all 54 genera of the Mucorales and Mortierellales based on combined analysis of actin and translation elongation factor EF-1alpha genes.

    PubMed

    Voigt, K; Wöstemeyer, J

    2001-05-30

    True fungi (Eumycota) are heterotrophic eukaryotic microorganisms encompassing ascomycetes, basidiomycetes, chytridiomycetes and zygomycetes. The natural systematics of the latter group, Zygomycota, are very poorly understood due to the lack of distinguishing morphological characters. We have determined sequences for the nuclear-encoded genes actin (act) from 82 zygomycetes representing all 54 currently recognized genera from the two zygomycetous orders Mucorales and Mortierellales. We also determined sequences for translation elongation factor EF-1alpha (tef) from 16 zygomycetes (total of 96,837 bp). Phylogenetic analysis in the context of available sequence data (total 2,062 nucleotide positions per species) revealed that current classification schemes for the mucoralean fungi are highly unnatural at the family and, to a large extent, at the genus level. The data clearly indicate a deep, ancient and distinct dichotomy of the orders Mucorales and Mortierellales, which are recognized only in some zygomycete systems. Yet at the same time the data show that two genera - Umbelopsis and Micromucor - previously placed within the Mortierellales on the basis of their weakly developed columella (a morphological structure of the sporangiophore well-developed within all Mucorales) are in fact members of the Mucorales. Phylogenetic analyses of the encoded amino acid sequences in the context of homologues from eukaryotes and archaebacterial outgroups indicate that the Eumycota studied here are a natural group but provide little or no support for the monophyly of either zygomycetes, ascomycetes or basidiomycetes. The data clearly indicate that a complete revision of zygomycete natural systematics is necessary. PMID:11404008

  6. Implications of Cellobiohydrolase Glycosylation for use in Biomass Conversion

    SciTech Connect

    Jeoh, T.; Michener W.; Himmel, M. E.; Decker, S. R.; Adney, W. S.

    2008-01-01

    The cellulase producing ascomycete, Trichoderma reesei (Hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. It is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. During the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. Another ascomycete, Aspergillus niger var. awamori, is also known to produce large quantities of heterologous proteins for industry. In this study, T. reesei Cel7A, a cellobiohydrolase, was expressed in A. niger var. awamori and subjected to detailed biophysical characterization. The purified recombinant enzyme contains six times the amount of N-linked glycan than the enzyme purified from a commercial T. reesei enzyme preparation. The activities of the two enzyme forms were compared using bacterial (microcrystalline) and phosphoric acid swollen (amorphous) cellulose as substrates. This comparison suggested that the increased level of N-glycosylation of the recombinant Cel7A (rCel7A) resulted in reduced activity and increased non-productive binding on cellulose. When treated with the N-glycosidase PNGaseF, the molecular weight of the recombinant enzyme approached that of the commercial enzyme and the activity on cellulose was improved.

  7. Identity and biodegradative abilities of yeasts isolated from plants growing in an arid climate.

    PubMed

    Middelhoven, W J

    1997-08-01

    Plants harvested in the Canary Islands Lanzarote and Fuerteventura were analyzed for the yeasts inhabiting their surface. Half of the isolates (22 out of 44) were identified as Debaryomyces hansenii. Black ascomycetes, viz. Hortaea werneckii and two Hormonema species were represented by 7 strains. Basidiomycetous yeasts, viz. Cryptococcus sp. (8 strains), Rhodotorula sp. (5 strains), Cerinosterus cyanescens (1 strain) and Pseudozyma sp. (1 strain) constituted a minority of 33%. Thirty strains were screened for their ability to assimilate various plant constituents including lipids of the cuticle and the cell membrane, hemicelluloses, nitrogenous compounds (protein, nucleic acids, amino acids) and benzene compounds. All strains were able to assimilate or to hydrolyze lipids, lecithin included. Many strains of D. hansenii, H. dematioides, H. werneckii, C. cyanescens, Cr. laurentii, Pseudozyma sp. and Rh. glutinis were proteolytic. Hemicelluloses like xylan and pectin were assimilated by black ascomycetous yeasts, Cryptococcus sp., Pseudozyma sp. and Rh. glutinis. Ferulic and hydroxycinnamic acids, gallic and tannic acids were assimilated by some strains of H. dematioides, C. cyanescens, Pseudozyma sp. and Rhodotorula sp. PMID:9298186

  8. Community composition of root-associated fungi in a Quercus-dominated temperate forest: “codominance” of mycorrhizal and root-endophytic fungi

    PubMed Central

    Toju, Hirokazu; Yamamoto, Satoshi; Sato, Hirotoshi; Tanabe, Akifumi S; Gilbert, Gregory S; Kadowaki, Kohmei

    2013-01-01

    In terrestrial ecosystems, plant roots are colonized by various clades of mycorrhizal and endophytic fungi. Focused on the root systems of an oak-dominated temperate forest in Japan, we used 454 pyrosequencing to explore how phylogenetically diverse fungi constitute an ecological community of multiple ecotypes. In total, 345 operational taxonomic units (OTUs) of fungi were found from 159 terminal-root samples from 12 plant species occurring in the forest. Due to the dominance of an oak species (Quercus serrata), diverse ectomycorrhizal clades such as Russula, Lactarius, Cortinarius, Tomentella, Amanita, Boletus, and Cenococcum were observed. Unexpectedly, the root-associated fungal community was dominated by root-endophytic ascomycetes in Helotiales, Chaetothyriales, and Rhytismatales. Overall, 55.3% of root samples were colonized by both the commonly observed ascomycetes and ectomycorrhizal fungi; 75.0% of the root samples of the dominant Q. serrata were so cocolonized. Overall, this study revealed that root-associated fungal communities of oak-dominated temperate forests were dominated not only by ectomycorrhizal fungi but also by diverse root endophytes and that potential ecological interactions between the two ecotypes may be important to understand the complex assembly processes of belowground fungal communities. PMID:23762515

  9. House dust mites possess a polymorphic, single domain putative peptidoglycan d,l endopeptidase belonging to the NlpC/P60 Superfamily.

    PubMed

    Tang, Vivian H; Stewart, Geoffrey A; Chang, Barbara J

    2015-01-01

    A 14 kDa protein homologous to the γ-d-glutamyl-l-diamino acid endopeptidase members of the NlpC/P60 Superfamily has been described in Dermatophagoides pteronyssinus and Dermatophagoides farinae but it is not clear whether other species produce homologues. Bioinformatics revealed homologous genes in other Sarcopteformes mite species (Psoroptes ovis and Blomia tropicalis) but not in Tetranychus urticae and Metaseiulus occidentalis. The degrees of identity (similarity) between the D. pteronyssinus mature protein and those from D. farinae, P. ovis and B. tropicalis were 82% (96%), 77% (93%) and 61% (82%), respectively. Phylogenetic studies showed the mite proteins were monophyletic and shared a common ancestor with both actinomycetes and ascomycetes. The gene encoding the D. pteronyssinus protein was polymorphic and intronless in contrast to that reported for D. farinae. Homology studies suggest that the mite, ascomycete and actinomycete proteins are involved in the catalysis of stem peptide attached to peptidoglycan. The finding of a gene encoding a P60 family member in the D. pteronyssinus genome together with the presence of a bacterial promotor suggests an evolutionary link to one or more prokaryotic endosymbionts. PMID:26566476

  10. Nucleotide Composition of Nucleic Acids of Fungi I. Ribonucleic Acids

    PubMed Central

    Storck, Roger

    1965-01-01

    Storck, Roger (The University of Texas, Austin). Nucleotide composition of nucleic acids from fungi. I. Ribonucleic acids. J. Bacteriol. 90:1260–1264. 1965.—The nucleotide composition of the ribonucleic acids (RNA) present in extracts of 26 species of fungi was determined. The results were analyzed, together with those in the literature. It was found that the content in moles per cent of guanine plus cytosine (GC content) varied from 44.1 to 60.5 in a distribution composed of 8 species of zygomycetes, 10 of ascomycetes, 11 of deuteromycetes, and 8 of basidiomycetes. The GC-content range and average were, respectively, 44.1 to 49.3 and 46.4 for the zygomycetes, 47.4 to 54.4 and 50.2 for the ascomycetes, 48.2 to 54.5 and 51.6 for the deuteromycetes, and 50.4 to 60.5 and 52.4 for the basidiomycetes. The GC content averaged 45.6 and ranged from 44.1 to 46.3 for four Mucor species. In addition, GC contents significantly lower than 50 were also encountered in some species of Hemiascomycetidae, suggesting that AT type RNA is not uncommon in fungi. It was proposed that the base composition of fungal RNA might have a taxonomic and phylogenetic significance. PMID:5848326

  11. House dust mites possess a polymorphic, single domain putative peptidoglycan d,l endopeptidase belonging to the NlpC/P60 Superfamily

    PubMed Central

    Tang, Vivian H.; Stewart, Geoffrey A.; Chang, Barbara J.

    2015-01-01

    A 14 kDa protein homologous to the γ-d-glutamyl-l-diamino acid endopeptidase members of the NlpC/P60 Superfamily has been described in Dermatophagoides pteronyssinus and Dermatophagoides farinae but it is not clear whether other species produce homologues. Bioinformatics revealed homologous genes in other Sarcopteformes mite species (Psoroptes ovis and Blomia tropicalis) but not in Tetranychus urticae and Metaseiulus occidentalis. The degrees of identity (similarity) between the D. pteronyssinus mature protein and those from D. farinae, P. ovis and B. tropicalis were 82% (96%), 77% (93%) and 61% (82%), respectively. Phylogenetic studies showed the mite proteins were monophyletic and shared a common ancestor with both actinomycetes and ascomycetes. The gene encoding the D. pteronyssinus protein was polymorphic and intronless in contrast to that reported for D. farinae. Homology studies suggest that the mite, ascomycete and actinomycete proteins are involved in the catalysis of stem peptide attached to peptidoglycan. The finding of a gene encoding a P60 family member in the D. pteronyssinus genome together with the presence of a bacterial promotor suggests an evolutionary link to one or more prokaryotic endosymbionts. PMID:26566476

  12. Cloning and expression in Pichia pastoris of an Irpex lacteus rhamnogalacturonan hydrolase tolerant to acetylated rhamnogalacturonan.

    PubMed

    Normand, J; Bonnin, E; Delavault, P

    2012-06-01

    In order to produce a recombinant rhamnogalacturonase from the basidiomycete Irpex lacteus using a molecular approach, PCR primers were designed based on a sequence alignment of four known ascomycete rhamnogalacturonases. Using 5' and 3' rapid amplification of cDNA ends (RACE) experiments, a 1,437-bp full-length cDNA containing an open reading frame of 1,329 bp was isolated. The corresponding putative protein sequence is of 443 amino acids and contains a secretion signal sequence of 22 amino acids. The theoretical mass of this protein is 44.6 kDa with a theoretical isoelectric point of 6.2. The amino acid sequence shared not only significant identities with ascomycete and basidiomycete putative rhamnogalacturonases but also complete similarity with peptides obtained from a recently purified rhamnogalacturonase from I. lacteus. The recombinant protein was successfully expressed in active form in Pichia pastoris. SDS-PAGE assay demonstrated that the recombinant enzyme was secreted in the culture medium and had a molar mass of 56 kDa. This recombinant rhamnogalacturonan hydrolase exhibited a pH optimum between 4.5 and 5 and a temperature optimum between 40C and 50C, which correspond to that of the native rhamnogalacturonase from I. lacteus. The study of its specificity through reaction products analysis showed that it was highly tolerant to the presence of acetyl groups on its substrate, even more than the native enzyme. PMID:22101785

  13. The reassignment of three 'lost' Taphrina species (Taphrina bullata, Taphrina insititiae and Taphrina rhizophora) supported by the divergence of nuclear and mitochondrial DNA.

    PubMed

    Petrydesová, Jana; Bacigálová, Kamila; Sulo, Pavol

    2013-08-01

    The ascomycetous genus Taphrina Fries originally contained more than 90 phytopathogenic microscopic dimorphic ascomycetes causing specific infections in different vascular plants. Although species have mainly been identified historically according to their host and morphological and physiological traits, they can be studied and preserved in the form of yeasts arising from germinating ascospores. Due to low DNA sequence divergence and the lack of available strains, the number of accepted Taphrina species has currently been reduced to 28. The aim of this study is the description of three previously accepted species. Taphrina bullata (type strain CCY 58-4-1 = CBS 12783), Taphrina insititiae (type strain CCY 58-5-1 = CBS 12782) and Taphrina rhizophora (type strain CCY 58-6-1 = CBS 12781), which have been omitted from a recent key. The host range, the divergence of the 26S rRNA gene sequence, internal transcribed spacers (ITS) and the mitochondrial small ribosomal subunit (rns) sequence strongly support their reassignment as species. PMID:23710051

  14. Plant-Polysaccharide-Degrading Enzymes from Basidiomycetes

    PubMed Central

    Rytioja, Johanna; Hildén, Kristiina; Yuzon, Jennifer; Hatakka, Annele; de Vries, Ronald P.

    2014-01-01

    SUMMARY Basidiomycete fungi subsist on various types of plant material in diverse environments, from living and dead trees and forest litter to crops and grasses and to decaying plant matter in soils. Due to the variation in their natural carbon sources, basidiomycetes have highly varied plant-polysaccharide-degrading capabilities. This topic is not as well studied for basidiomycetes as for ascomycete fungi, which are the main sources of knowledge on fungal plant polysaccharide degradation. Research on plant-biomass-decaying fungi has focused on isolating enzymes for current and future applications, such as for the production of fuels, the food industry, and waste treatment. More recently, genomic studies of basidiomycete fungi have provided a profound view of the plant-biomass-degrading potential of wood-rotting, litter-decomposing, plant-pathogenic, and ectomycorrhizal (ECM) basidiomycetes. This review summarizes the current knowledge on plant polysaccharide depolymerization by basidiomycete species from diverse habitats. In addition, these data are compared to those for the most broadly studied ascomycete genus, Aspergillus, to provide insight into specific features of basidiomycetes with respect to plant polysaccharide degradation. PMID:25428937

  15. Peptaibol production by sepedonium strains parasitizing boletales.

    PubMed

    Neuhof, Torsten; Berg, Albrecht; Besl, Helmut; Schwecke, Torsten; Dieckmann, Ralf; von Döhren, Hans

    2007-06-01

    Fungi of the genus Sepedonium (anamorphic ascomycetes) are known to infect fruiting bodies of Basidiomycetes of the order Boletales. We have characterized twelve Sepedonium isolates by intact-cell mass spectrometry (IC-MS) with the help of respective biomarkers and their metabolite spectra focusing on peptaibol production. A strain of mycoparasitic S. chalcipori was grown in solid-state fermentation, and tylopeptin production was found, suggesting an ascomycete origin of these peptaibols, which were first described in the basidiomycete Tylopilus neofelleus. In addition, the structures of two new peptaibols, chalciporin A (=Ac-Trp-Val-Aib-Val-Ala-Gln-Ala-Aib-Ser-Leu-Ala-Leu-Aib-Gln-Leuol) and chalciporin B (=Ac-Trp-Val-Aib-Val-Ala-Gln-Ala-Aib-Gln-Aib-Ala-Leu-Aib-Gln-Leuol) are presented. The IC-MS technique was applied for in situ peptaibol analysis of Sepedonium strains growing on Boletales, in particular S. chrysospermum infecting Xerocomus cf. badius. We found chrysospermins at the surface and within basidiomycete tissue, as well as in the cultivated parasite. PMID:17589879

  16. The tempo and modes of evolution of reproductive isolation in fungi

    PubMed Central

    Giraud, T; Gourbière, S

    2012-01-01

    Reproductive isolation is an essential ingredient of speciation, and much has been learned in recent years about the evolution of reproductive isolation and the genetics of reproductive barriers in animals and plants. Fungi have been neglected on these aspects, despite being tractable model eukaryotes. Here, we used a model fitting approach to look at the importance of different barriers to gene flow to explain the decrease of reproductive compatibility with genetic distance in fungi. We found support for the occurrence of reinforcement in the presyngamy compatibility among basidiomycetes. In contrast, no evidence for reinforcement was detected in ascomycetes, concurring with the idea that host/habitat adaptation in this group can pleiotropically cause reproductive isolation. We found no evidence of a snowballing accumulation of postsyngamic reproductive incompatibilities in either ascomycetes or the complex of anther smut fungi. Together with previous studies, our results suggest that ecologically based barriers to gene flow and karyotypic differences may have an important role in hybrid inviability and sterility in fungi. Interestingly, hybrid sterility appeared to evolve faster than hybrid inviability in fungi. PMID:22669076

  17. Protoplasmic organization of hyphal tips among fungi: vesicles and Spitzenkrper.

    PubMed

    Grove, S N; Bracker, C E

    1970-11-01

    Hyphal tips of fungi representing Omycetes, Zygomycetes, Ascomycetes, Basidiomycetes, and Deuteromycetes were examined by light and electron microscopy and compared with respect to their protoplasmic organization. In all fungi studied, there is a zone at the hyphal apex which is rich in cytoplasmic vesicles but nearly devoid of other cell components. Some vesicle profiles are continuous with the plasma membrane at the apices of these tip-growing cells. The subapical zones of hyphae contain an endomembrane system which includes smooth-surfaced cisternae associated with small clusters of vesicles. The findings are consistent with the hypothesis that vesicles produced by the endomembrane system in the subapical region become concentrated in the apex where they are incorporated at the expanding surface. Septate fungi (Ascomycetes, Basidiomycetes, and Deuteromycetes) have an apical body (Spitzenkrper) which is associated with growing hyphal tips. In electron micrographs of these fungi, an additional specialized region within the accumulation of apical vesicles is shown for the first time. This region corresponds on the bases of distribution among fungi, location in hyphae, size, shape and boundary characteristics to the Spitzenkrper seen by light microscopy. This structure is not universally associated with tip growth, whereas apical vesicles are widespread among tip-growing systems. PMID:4099103

  18. Scheffersomyces cryptocercus: a new xylose-fermenting yeast associated with the gut of wood roaches and new combinations in the Sugiyamaella yeast clade.

    PubMed

    Urbina, Hector; Frank, Robert; Blackwell, Meredith

    2013-01-01

    The gut of wood-feeding insects is a microhabitat for a specialized community of microbes, including bacteria and several groups of eukaryotes such as nematodes, parabasalids and fungi. The characterization of gut yeast communities from a variety of insects has shown that certain yeasts often are associated with the insects. The gut of wood-feeding insects is rich in ascomycete yeasts and in particular xylose-fermenting (X-F) and assimilating yeasts have been consistently present in the gut of lignicolous insects. The objective of this study was the characterization of the yeast flora from the gut of the wood roach Cryptocercus sp. (Blattodea: Cryptocercidae). Five wood roaches were collected along the Appalachian Trail near the border between Tennessee and North Carolina, USA. We isolated 18 yeast strains from the wood roaches identified as Sugiyamaella paludigena and Sugiyamaella lignohabitans, xylose-assimilating yeasts, and Scheffersomyces cryptocercus (NRRL Y-48824(T) = CBS 12658) a new species of X-F yeast. The presence of X-F and certain non X-F yeasts in the gut of the subsocial wood roach Cryptocercus sp. extends the previous findings of associations between certain ascomycete yeasts and lignicolous insects. New combinations were made for 13 asexual members of the Sugiyamaella clade. PMID:23233509

  19. The Ustilago maydis Nit2 homolog regulates nitrogen utilization and is required for efficient induction of filamentous growth.

    PubMed

    Horst, Robin J; Zeh, Christine; Saur, Alexandra; Sonnewald, Sophia; Sonnewald, Uwe; Voll, Lars M

    2012-03-01

    Nitrogen catabolite repression (NCR) is a regulatory strategy found in microorganisms that restricts the utilization of complex and unfavored nitrogen sources in the presence of favored nitrogen sources. In fungi, this concept has been best studied in yeasts and filamentous ascomycetes, where the GATA transcription factors Gln3p and Gat1p (in yeasts) and Nit2/AreA (in ascomycetes) constitute the main positive regulators of NCR. The reason why functional Nit2 homologs of some phytopathogenic fungi are required for full virulence in their hosts has remained elusive. We have identified the Nit2 homolog in the basidiomycetous phytopathogen Ustilago maydis and show that it is a major, but not the exclusive, positive regulator of nitrogen utilization. By transcriptome analysis of sporidia grown on artificial media devoid of favored nitrogen sources, we show that only a subset of nitrogen-responsive genes are regulated by Nit2, including the Gal4-like transcription factor Ton1 (a target of Nit2). Ustilagic acid biosynthesis is not under the control of Nit2, while nitrogen starvation-induced filamentous growth is largely dependent on functional Nit2. nit2 deletion mutants show the delayed initiation of filamentous growth on maize leaves and exhibit strongly compromised virulence, demonstrating that Nit2 is required to efficiently initiate the pathogenicity program of U. maydis. PMID:22247264

  20. Zygomycetes in Vesicular Basanites from Vesteris Seamount, Greenland Basin – A New Type of Cryptoendolithic Fungi

    PubMed Central

    Ivarsson, Magnus; Peckmann, Jörn; Tehler, Anders; Broman, Curt; Bach, Wolfgang; Behrens, Katharina; Reitner, Joachim; Böttcher, Michael E.; Norbäck Ivarsson, Lena

    2015-01-01

    Fungi have been recognized as a frequent colonizer of subseafloor basalt but a substantial understanding of their abundance, diversity and ecological role in this environment is still lacking. Here we report fossilized cryptoendolithic fungal communities represented by mainly Zygomycetes and minor Ascomycetes in vesicles of dredged volcanic rocks (basanites) from the Vesteris Seamount in the Greenland Basin. Zygomycetes had not been reported from subseafloor basalt previously. Different stages in zygospore formation are documented in the studied samples, representing a reproduction cycle. Spore structures of both Zygomycetes and Ascomycetes are mineralized by romanechite-like Mn oxide phases, indicating an involvement in Mn(II) oxidation to form Mn(III,VI) oxides. Zygospores still exhibit a core of carbonaceous matter due to their resistance to degradation. The fungi are closely associated with fossiliferous marine sediments that have been introduced into the vesicles. At the contact to sediment infillings, fungi produced haustoria that penetrated and scavenged on the remains of fragmented marine organisms. It is most likely that such marine debris is the main carbon source for fungi in shallow volcanic rocks, which favored the establishment of vital colonies. PMID:26181773

  1. Rhizosphere communities of genetically modified zeaxanthin-accumulating potato plants and their parent cultivar differ less than those of different potato cultivars.

    PubMed

    Weinert, Nicole; Meincke, Remo; Gottwald, Christine; Heuer, Holger; Gomes, Newton C M; Schloter, Michael; Berg, Gabriele; Smalla, Kornelia

    2009-06-01

    The effects of genetically modified (GM), zeaxanthin-accumulating potato plants on microbial communities in the rhizosphere were compared to the effects of different potato cultivars. Two GM lines and their parental cultivar, as well as four other potato cultivars, were grown in randomized field plots at two sites and in different years. Rhizosphere samples were taken at three developmental stages during plant growth and analyzed using denaturing gradient gel electrophoresis (DGGE) fingerprints of Bacteria, Actinobacteria, Alpha- and Betaproteobacteria, Bacillus, Streptomycetaceae, Pseudomonas, gacA, Fungi, and Ascomycetes. In the bacterial DGGE gels analyzed, significant differences between the parental cultivar and the two GM lines were detected mainly for Actinobacteria but also for Betaproteobacteria and Streptomycetaceae, yet these differences occurred only at one site and in one year. Significant differences occurred more frequently for Fungi, especially Ascomycetes, than for bacteria. When all seven plant genotypes were compared, DGGE analysis revealed that different cultivars had a greater effect on both bacterial and fungal communities than genetic modification. The effects of genetic modification were detected mostly at the senescence developmental stage of the plants. The site was the overriding factor affecting microbial community structure compared to the plant genotype. In general, the fingerprints of the two GM lines were more similar to that of the parental cultivar, and the differences observed did not exceed natural cultivar-dependent variability. PMID:19376893

  2. Antifungal Susceptibility Profiles of 1698 Yeast Reference Strains Revealing Potential Emerging Human Pathogens

    PubMed Central

    Desnos-Ollivier, Marie; Robert, Vincent; Raoux-Barbot, Dorothée; Groenewald, Marizeth; Dromer, Françoise

    2012-01-01

    New molecular identification techniques and the increased number of patients with various immune defects or underlying conditions lead to the emergence and/or the description of novel species of human and animal fungal opportunistic pathogens. Antifungal susceptibility provides important information for ecological, epidemiological and therapeutic issues. The aim of this study was to assess the potential risk of the various species based on their antifungal drug resistance, keeping in mind the methodological limitations. Antifungal susceptibility profiles to the five classes of antifungal drugs (polyens, azoles, echinocandins, allylamines and antimetabolites) were determined for 1698 yeast reference strains belonging to 992 species (634 Ascomycetes and 358 Basidiomycetes). Interestingly, geometric mean minimum inhibitory concentrations (MICs) of all antifungal drugs tested were significantly higher for Basidiomycetes compared to Ascomycetes (p<0.001). Twenty four strains belonging to 23 species of which 19 were Basidiomycetes seem to be intrinsically “resistant” to all drugs. Comparison of the antifungal susceptibility profiles of the 4240 clinical isolates and the 315 reference strains belonging to 53 shared species showed similar results. Even in the absence of demonstrated in vitro/in vivo correlation, knowing the in vitro susceptibility to systemic antifungal agents and the putative intrinsic resistance of yeast species present in the environment is important because they could become opportunistic pathogens. PMID:22396754

  3. Mycological evidence of coprophagy from the feces of an Alaskan Late Glacial mammoth

    NASA Astrophysics Data System (ADS)

    van Geel, Bas; Guthrie, R. Dale; Altmann, Jens G.; Broekens, Peter; Bull, Ian D.; Gill, Fiona L.; Jansen, Boris; Nieman, Aline M.; Gravendeel, Barbara

    2011-08-01

    Dung from a mammoth was preserved under frozen conditions in Alaska. The mammoth lived during the early part of the Late Glacial interstadial (ca 12,300 BP). Microfossils, macroremains and ancient DNA from the dung were studied and the chemical composition was determined to reconstruct both the paleoenvironment and paleobiology of this mammoth. Pollen spectra are dominated by Poaceae, Artemisia and other light-demanding taxa, indicating an open, treeless landscape ('mammoth steppe'). Fruits and seeds support this conclusion. The dung consists mainly of cyperaceous stems and leaves, with a minor component of vegetative remains of Poaceae. Analyses of fragments of the plastid rbcL gene and trnL intron and nrITS1 region, amplified from DNA extracted from the dung, supplemented the microscopic identifications. Many fruit bodies with ascospores of the coprophilous fungus Podospora conica were found inside the dung ball, indicating that the mammoth had eaten dung. The absence of bile acids points to mammoth dung. This is the second time that evidence for coprophagy of mammoths has been derived from the presence of fruit bodies of coprophilous fungi in frozen dung. Coprophagy might well have been a common habit of mammoths. Therefore, we strongly recommend that particular attention should be given to fungal remains in future fossil dung studies.

  4. DNA barcoding and phylogenetic relationships in Anatidae.

    PubMed

    Huang, Zuhao; Yang, Chengzhong; Ke, Dianhua

    2016-01-01

    Mitochondrial cytochrome c oxidase subunit I (COI) has been used as a powerful marker in a variety of phylogenetic studies. According to studies of bird species, the 694-bp sequence of the mitochondrial gene encoding COI is extremely useful for species identification and phylogeny. In the present study, we analyzed the COI barcodes of 79 species from 26 genera belonging to the Anatidae family. Sixty-six species (83.54%) of the species were identified correctly from their DNA barcodes. The remaining 13 species shared barcodes sequences with closely related species. Kimura two-parameter (K2P) distances were calculated between barcodes. The average genetic distance between species was 41 times higher compared to the average genetic distance within species. Neighbor-joining method was used to construct a phylogenetic tree, which grouped all of the genera into three divergent clades. Dendrocygna and Nomonyx + Oxyura were identified as early offshoots of the Anatidae. All the remaining taxa fell into two clades that correspond to the two subfamilies Anserinae and Anatiane. Based on our results, DNA barcoding is an effective molecular tool for Anatidae species identification and phylogenetic inference. PMID:24938090

  5. Microsatellites identify depredated waterfowl remains from glaucous gull stomachs

    USGS Publications Warehouse

    Scribner, K.T.; Bowman, T.D.

    1998-01-01

    Prey remains can provide valuable sources of information regarding causes of predation and the species composition of a predator's diet. Unfortunately, the highly degraded state of many prey samples from gastrointestinal tracts often precludes unambiguous identification. We describe a procedure by which PCR amplification of taxonomically informative microsatellite loci were used to identify species of waterfowl predated by glaucous gulls (Larus hyperboreus). We found that one microsatellite locus unambiguously distinguished between species of the subfamily Anserinae (whistling ducks, geese and swans) and those of the subfamily Anatidae (all other ducks). An additional locus distinguished the remains of all geese and swan species known to nest on the Yukon-Kuskokwim delta in western Alaska. The study focused on two waterfowl species which have experienced precipitous declines in population numbers: emperor geese (Chen canagica) and spectacled eiders (Somateria fischeri). No evidence of predation on spectacled eiders was observed. Twenty-six percent of all glaucous gull stomachs examined contained the remains of juvenile emperor geese.

  6. Communities of endophytic sebacinales associated with roots of herbaceous plants in agricultural and grassland ecosystems are dominated by Serendipita herbamans sp. nov.

    PubMed

    Riess, Kai; Oberwinkler, Franz; Bauer, Robert; Garnica, Sigisfredo

    2014-01-01

    Endophytic fungi are known to be commonly associated with herbaceous plants, however, there are few studies focusing on their occurrence and distribution in plant roots from ecosystems with different land uses. To explore the phylogenetic diversity and community structure of Sebacinales endophytes from agricultural and grassland habitats under different land uses, we analysed the roots of herbaceous plants using strain isolation, polymerase chain reaction (PCR), transmission electron microscopy (TEM) and co-cultivation experiments. A new sebacinoid strain named Serendipita herbamans belonging to Sebacinales group B was isolated from the roots of Bistorta vivipara, which is characterized by colourless monilioid cells (chlamydospores) that become yellow with age. This species was very common and widely distributed in association with a broad spectrum of herbaceous plant families in diverse habitats, independent of land use type. Ultrastructurally, the presence of S. herbamans was detected in the cortical cells of Plantago media, Potentilla anserina and Triticum aestivum. In addition, 13 few frequent molecular operational taxonomic units (MOTUs) or species were found across agricultural and grassland habitats, which did not exhibit a distinctive phylogenetic structure. Laboratory-based assays indicate that S. herbamans has the ability to colonize fine roots and stimulate plant growth. Although endophytic Sebacinales are widely distributed across agricultural and grassland habitats, TEM and nested PCR analyses reinforce the observation that these microorganisms are present in low quantity in plant roots, with no evidence of host specificity. PMID:24743185

  7. Identification of a novel bile acid in swans, tree ducks, and geese: 3alpha,7alpha,15alpha-trihydroxy-5beta-cholan-24-oic acid.

    PubMed

    Kakiyama, Genta; Iida, Takashi; Goto, Takaaki; Mano, Nariyasu; Goto, Junichi; Nambara, Toshio; Hagey, Lee R; Schteingart, Claudio D; Hofmann, Alan F

    2006-07-01

    By HPLC, a taurine-conjugated bile acid with a retention time different from that of taurocholate was found to be present in the bile of the black-necked swan, Cygnus melanocoryphus. The bile acid was isolated and its structure, established by (1)H and (13)C NMR and mass spectrometry, was that of the taurine N-acyl amidate of 3alpha,7alpha,15alpha-trihydroxy-5beta-cholan-24-oic acid. The compound was shown to have chromatographic and spectroscopic properties that were identical to those of the taurine conjugate of authentic 3alpha,7alpha,15alpha-trihydroxy-5beta-cholan-24-oic acid, previously synthesized by us from ursodeoxycholic acid. By HPLC, the taurine conjugate of 3alpha,7alpha,15alpha-trihydroxy-5beta-cholan-24-oic acid was found to be present in 6 of 6 species in the subfamily Dendrocygninae (tree ducks) and in 10 of 13 species in the subfamily Anserinae (swans and geese) but not in other subfamilies in the Anatidae family. It was also not present in species from the other two families of the order Anseriformes. 3alpha,7alpha,15alpha-Trihydroxy-5beta-cholan-24-oic acid is a new primary bile acid that is present in the biliary bile acids of swans, tree ducks, and geese and may be termed 15alpha-hydroxy-chenodeoxycholic acid. PMID:16648547

  8. Two siblings with triple A syndrome and novel mutation presenting as hereditary polyneuropathy.

    PubMed

    Dumić, Miroslav; Barišić, Nina; Rojnić-Putarek, Nataša; Kušec, Vesna; Stanimirović, Andrija; Koehler, Katrin; Huebner, Angela

    2011-03-01

    The clinical and molecular data on triple A syndrome in two siblings (girl 3.5 years and boy 5.5 years at presentation) with early onset of neurological dysfunction are described. Both patients showed delayed developmental milestones and neurological dysfunctions (motor and sensory demyelinating neuropathy, marked hyperreflexia, calves hypothrophy, pes cavus, gait disturbance) in early childhood, when erroneously diagnosed with hereditary polyneuropathy, most likely Charcot-Marie-Tooth disease. After a severe adrenal crisis in the younger sister at the age of 3 years, the older brother aged 5.5 years was also evaluated and latent adrenal insufficiency was discovered. As both of the siblings had alacrima, hyperkeratosis of palms, cutis anserina, and nasal speech, diagnosis of triple A syndrome was considered. Sequencing of the AAAS gene detected a compound heterozygous mutation consisting of a novel mutation p.Ser296Tyr (c.887C>A) in exon 9 and a previously described p.Ser263Pro (c.787T>C) missense mutation in exon 8 in both siblings. In conclusion, triple A syndrome should be considered in patients presenting with early neurological dysfunction and developmental delay. Alacrima as the earliest and most consistent clinical sign should be investigated by Schirmer test. Patients should be regularly tested for adrenal dysfunction to prevent life-threatening adrenal crises. PMID:20931227

  9. Overwintering of Vineyard Yeasts: Survival of Interacting Yeast Communities in Grapes Mummified on Vines

    PubMed Central

    Sipiczki, Matthias

    2016-01-01

    The conversion of grape must into wine involves the development and succession of yeast populations differing in species composition. The initial population is formed by vineyard strains which are washed into the must from the crushed grapes and then completed with yeasts coming from the cellar environment. As the origin and natural habitat of the vineyard yeasts are not fully understood, this study addresses the possibility, that grape yeasts can be preserved in berries left behind on vines at harvest until the spring of the next year. These berries become mummified during the winter on the vines. To investigate whether yeasts can survive in these overwintering grapes, mummified berries were collected in 16 localities in the Tokaj wine region (Hungary-Slovakia) in early March. The collected berries were rehydrated to recover viable yeasts by plating samples onto agar plates. For the detection of minority species which would not be detected by direct plating, an enrichment step repressing the propagation of alcohol-sensitive yeasts was also included in the process. The morphological, physiological, and molecular analysis identified 13 basidiomycetous and 23 ascomycetous species including fermentative yeasts of wine-making relevance among the 3879 isolates. The presence of viable strains of these species demonstrates that the grapes mummified on the vine can serve as a safe reservoir of yeasts, and may contribute to the maintenance of grape-colonizing yeast populations in the vineyard over years, parallel with other vectors and habitats. All basidiomycetous species were known phylloplane yeasts. Three Hanseniaspora species and pigmented Metschnikowia strains were the most frequent ascomycetes. Other fermentative yeasts of wine-making relevance were detected only in the enrichment cultures. Saccharomyces (S. paradoxus, S. cerevisiae, and S. uvarum) were recovered from 13% of the samples. No Candida zemplinina was found. The isolates with Aureobasidium morphology turned out to belong to Aureobasidium subglaciale, Kabatiella microsticta, or Columnosphaeria fagi. The ascomyceteous isolates grew at high concentrations of sugars with Wickerhamomyces anomalus being the most tolerant species. Complex interactions including antagonism (growth inhibition, contact inhibition, competition for nutrients) and synergism (crossfeeding) among the isolates and with Botrytis cinerea shape the composition of the overwintering communities. PMID:26973603

  10. Ecological study of the fungal populations of the acidic Tinto River in southwestern Spain.

    PubMed

    López-Archilla, A I; González, A E; Terrón, M C; Amils, R

    2004-11-01

    The characterization of the microbial ecology of the Tinto River, an extreme habitat with an extremely low pH and a high concentration of heavy metals, revealed an unexpected level of microbial richness. A variety of microbial eukaryotes was isolated, among them several fungal strains that were identified and their physiological characteristics studied. Ninety strains of yeast were isolated from the Tinto River. Fifty-two percent of them were capable of growth in vitro using medium amended with river water. They belong to 6 genera of basidiomycetes (Rhodotorula, Cryptococcus, Tremella, Holtermannia, Leucosporidium, and Mrakia) and 2 of ascomycetes (Candida and Williopsis). In addition, 349 strains of hyphomycetes belonging to 17 genera (most of them ascomycetes) were isolated and studied. Forty-four percent of the isolated filamentous fungi (154 strains) were capable of growing in vitro using medium amended with Tinto River water. Of this percentage, 19% (29 strains) belonged to the genus Penicillium (16 species) and 66% (102 strains) were included in the genera Scytalidium, Bahusakala, Phoma, and Heteroconium or showed dark sterile mycelia, which probably are of dematiaceous hyphomycetes. In addition, we characterized strains of the ascomycete genera Lecythophora and Acremonium and of the zygomycete genus Mortierella, all of them capable of growing in medium amended with river water. Statistical correlation of biological and physicochemical variables suggested a positive relationship between the dematiaceous hyphomycetes and the most extreme physicochemical conditions found in the Tinto River. Principal components analysis confirmed this relationship and also showed that the Acremonium and Lecythophora groups had environmental preferences similar to those of dematiaceous fungi. The spatial positions of the sampling sites were grouped in 2 main clusters: (i) sampling sites in the mine zone in which most of the dematiaceous, Acremonium, and Lecythophora strains were isolated and (ii) sites that were not in the mine zone and sampling station 5 from which were isolated mainly strains of fungi that were not capable of growing in the medium amended with river water and species of the Penicillium genus. PMID:15644909

  11. Overwintering of Vineyard Yeasts: Survival of Interacting Yeast Communities in Grapes Mummified on Vines.

    PubMed

    Sipiczki, Matthias

    2016-01-01

    The conversion of grape must into wine involves the development and succession of yeast populations differing in species composition. The initial population is formed by vineyard strains which are washed into the must from the crushed grapes and then completed with yeasts coming from the cellar environment. As the origin and natural habitat of the vineyard yeasts are not fully understood, this study addresses the possibility, that grape yeasts can be preserved in berries left behind on vines at harvest until the spring of the next year. These berries become mummified during the winter on the vines. To investigate whether yeasts can survive in these overwintering grapes, mummified berries were collected in 16 localities in the Tokaj wine region (Hungary-Slovakia) in early March. The collected berries were rehydrated to recover viable yeasts by plating samples onto agar plates. For the detection of minority species which would not be detected by direct plating, an enrichment step repressing the propagation of alcohol-sensitive yeasts was also included in the process. The morphological, physiological, and molecular analysis identified 13 basidiomycetous and 23 ascomycetous species including fermentative yeasts of wine-making relevance among the 3879 isolates. The presence of viable strains of these species demonstrates that the grapes mummified on the vine can serve as a safe reservoir of yeasts, and may contribute to the maintenance of grape-colonizing yeast populations in the vineyard over years, parallel with other vectors and habitats. All basidiomycetous species were known phylloplane yeasts. Three Hanseniaspora species and pigmented Metschnikowia strains were the most frequent ascomycetes. Other fermentative yeasts of wine-making relevance were detected only in the enrichment cultures. Saccharomyces (S. paradoxus, S. cerevisiae, and S. uvarum) were recovered from 13% of the samples. No Candida zemplinina was found. The isolates with Aureobasidium morphology turned out to belong to Aureobasidium subglaciale, Kabatiella microsticta, or Columnosphaeria fagi. The ascomyceteous isolates grew at high concentrations of sugars with Wickerhamomyces anomalus being the most tolerant species. Complex interactions including antagonism (growth inhibition, contact inhibition, competition for nutrients) and synergism (crossfeeding) among the isolates and with Botrytis cinerea shape the composition of the overwintering communities. PMID:26973603

  12. Essential Gene Discovery in the Basidiomycete Cryptococcus neoformans for Antifungal Drug Target Prioritization

    PubMed Central

    Ianiri, Giuseppe

    2015-01-01

    ABSTRACT Fungal diseases represent a major burden to health care globally. As with other pathogenic microbes, there is a limited number of agents suitable for use in treating fungal diseases, and resistance to these agents can develop rapidly. Cryptococcus neoformans is a basidiomycete fungus that causes cryptococcosis worldwide in both immunocompromised and healthy individuals. As a basidiomycete, it diverged from other common pathogenic or model ascomycete fungi more than 500 million years ago. Here, we report C. neoformans genes that are essential for viability as identified through forward and reverse genetic approaches, using an engineered diploid strain and genetic segregation after meiosis. The forward genetic approach generated random insertional mutants in the diploid strain, the induction of meiosis and sporulation, and selection for haploid cells with counterselection of the insertion event. More than 2,500 mutants were analyzed, and transfer DNA (T-DNA) insertions in several genes required for viability were identified. The genes include those encoding the thioredoxin reductase (Trr1), a ribosome assembly factor (Rsa4), an mRNA-capping component (Cet1), and others. For targeted gene replacement, the C. neoformans homologs of 35 genes required for viability in ascomycete fungi were disrupted, meiosis and sporulation were induced, and haploid progeny were evaluated for their ability to grow on selective media. Twenty-one (60%) were found to be required for viability in C. neoformans. These genes are involved in mitochondrial translation, ergosterol biosynthesis, and RNA-related functions. The heterozygous diploid mutants were evaluated for haploinsufficiency on a number of perturbing agents and drugs, revealing phenotypes due to the loss of one copy of an essential gene in C. neoformans. This study expands the knowledge of the essential genes in fungi using a basidiomycete as a model organism. Genes that have no mammalian homologs and are essential in both Cryptococcus and ascomycete human pathogens would be ideal for the development of antifungal drugs with broad-spectrum activity. PMID:25827419

  13. High diversity and complex evolution of fungal cytochrome P450 reductase: cytochrome P450 systems.

    PubMed

    Lah, Ljerka; Krasevec, Nada; Trontelj, Peter; Komel, Radovan

    2008-04-01

    Cytochrome P450 reductase (CPR) is the redox partner of P450 monooxygenases, involved in primary and secondary metabolism of eukaryotes. Two novel CPR genes, sharing 34% amino acid identity, were found in the filamentous ascomycete Cochliobolus lunatus. Fungal genomes were searched for putative CPR enzymes. Phylogenetic analysis suggests that multiple independent CPR duplication events occurred in fungi, whereas P450-CPR fusion occurred before the diversification of Dikarya and Zygomycota. Additionally, losses of methionine synthase reductase were found in certain fungal taxa; a truncated form of this enzyme was conserved in Pezizomycotina. In fungi, high numbers of cytochrome P450 enzymes, multiple CPRs, and P450-CPR fusion proteins were associated with filamentous growth. Evolution of multiple CPR-like oxidoreductases in filamentous fungi might have been driven by the complexity of biochemical functions necessitated by their growth form, as opposed to yeast. PMID:18024101

  14. Draft Genome Sequence of Colletotrichum falcatum - A Prelude on Screening of Red Rot Pathogen in Sugarcane

    PubMed Central

    Viswanathan, Rasappa; Prasanth, Chandrasekaran Naveen; Malathi, Palaniyandi; Sundar, Amalraj Ramesh

    2016-01-01

    Colletotrichum falcatum, a concealed fungal ascomycete causes red rot, which is a serious disease in sugarcane. It infects economically important stalk tissues, considered as store house of sugar in sugarcane. The study is to find genetic complexities of C. falcatum in establishing this as a stalk infecting pathogen and to decipher the unique lifestyle of this pathogen using NGS technology. We report the draft genome of C. falcatum of about 48.16 Mb in size with 12,270 genes. The genome sequences were compared with other fungal species which revealed that C. falcatum is closely related to C. graminicola and C.sublineola the causal organisms of anthracnose in maize and sorghum. These results brought a new revelation to explore the lifestyle of this unique pathogen which is specialized to infect sugarcane stalk tissues in detail. PMID:26958090

  15. Fungal genomes mining to discover novel sterol esterases and lipases as catalysts

    PubMed Central

    2013-01-01

    Background Sterol esterases and lipases are enzymes able to efficiently catalyze synthesis and hydrolysis reactions of both sterol esters and triglycerides and due to their versatility could be widely used in different industrial applications. Lipases with this ability have been reported in the yeast Candida rugosa that secretes several extracellular enzymes with a high level of sequence identity, although different substrate specificity. This versatility has also been found in the sterol esterases from the ascomycetes Ophiostoma piceae and Melanocarpus albomyces. Results In this work we present an in silico search of new sterol esterase and lipase sequences from the genomes of environmental fungi. The strategy followed included identification and search of conserved domains from these versatile enzymes, phylogenetic studies, sequence analysis and 3D modeling of the selected candidates. Conclusions Six potential putative enzymes were selected and their kinetic properties and substrate selectivity are discussed on the basis of their similarity with previously characterized sterol esterases/lipases with known structures. PMID:24138290

  16. Evolution of regulatory networks in Candida glabrata: learning to live with the human host.

    PubMed

    Roy, Sushmita; Thompson, Dawn

    2015-12-01

    The opportunistic human fungal pathogen Candida glabrata is second only to C. albicans as the cause of Candida infections and yet is more closely related to Saccharomyces cerevisiae. Recent advances in functional genomics technologies and computational approaches to decipher regulatory networks, and the comparison of these networks among these and other Ascomycete species, have revealed both unique and shared strategies in adaptation to a human commensal/opportunistic pathogen lifestyle and antifungal drug resistance in C. glabrata. Recently, several C. glabrata sister species in the Nakeseomyces clade representing both human associated (commensal) and environmental isolates have had their genomes sequenced and analyzed. This has paved the way for comparative functional genomics studies to characterize the regulatory networks in these species to identify informative patterns of conservation and divergence linked to phenotypic evolution in the Nakaseomyces lineage. PMID:26449820

  17. The fungal teratogen secalonic acid D is an inhibitor of protein kinase C and of cyclic AMP-dependent protein kinase.

    PubMed

    Wang, B H; Polya, G M

    1996-04-01

    The teratogenic metabolite secalonic acid D deriving from the ergot-producing, rye-infecting ascomycete fungus Claviceps purpurea and from Penicillum oxalicum is an inhibitor of Ca2+- and phospholipid-dependent protein kinase C (PKC) and of the catalytic subunit of cyclic AMP-dependent protein kinase (cAK) (C50 values 15 microM and 12 microM, respectively). Secalonic acid D also inhibits Ca2+-calmodulin-dependent myosin light chain kinase (MLCK) and plant Ca2+-dependent protein kinase (CDPK). The inhibition of cAK by secalonic acid D is competitive with respect to both peptide substrate and ATP. However, secalonic acid D does not inhibit a high-affinity nucleotide-binding phosphatase from potato. A variety of other naturally-occurring teratogenic agents are not inhibitors of the protein kinases examined. PMID:8657739

  18. Untargeted Metabolic Profiling of Winery-Derived Biomass Waste Degradation by Penicillium chrysogenum.

    PubMed

    Karpe, Avinash V; Beale, David J; Godhani, Nainesh B; Morrison, Paul D; Harding, Ian H; Palombo, Enzo A

    2015-12-16

    Winery-derived biomass waste was degraded by Penicillium chrysogenum under solid state fermentation over 8 days in a (2)H2O-supplemented medium. Multivariate statistical analysis of the gas chromatography-mass spectrometry (GC-MS) data resulted in the identification of 94 significant metabolites, within 28 different metabolic pathways. The majority of biomass sugars were utilized by day 4 to yield products such as sugars, fatty acids, isoprenoids, and amino acids. The fungus was observed to metabolize xylose to xylitol, an intermediate of ethanol production. However, enzyme inhibition and autolysis were observed from day 6, indicating 5 days as the optimal time for fermentation. P. chrysogenum displayed metabolism of pentoses (to alcohols) and degraded tannins and lignins, properties that are lacking in other biomass-degrading ascomycetes. Rapid fermentation (3-5 days) may not only increase the pentose metabolizing efficiency but also increase the yield of medicinally important metabolites, such as syringate. PMID:26611372

  19. Genomic analyses and expression evaluation of thaumatin-like gene family in the cacao fungal pathogen Moniliophthora perniciosa.

    PubMed

    Franco, Sulamita de Freitas; Baroni, Renata Moro; Carazzolle, Marcelo Falsarella; Teixeira, Paulo José Pereira Lima; Reis, Osvaldo; Pereira, Gonçalo Amarante Guimarães; Mondego, Jorge Maurício Costa

    2015-10-30

    Thaumatin-like proteins (TLPs) are found in diverse eukaryotes. Plant TLPs, known as Pathogenicity Related Protein (PR-5), are considered fungal inhibitors. However, genes encoding TLPs are frequently found in fungal genomes. In this work, we have identified that Moniliophthora perniciosa, a basidiomycete pathogen that causes the Witches' Broom Disease (WBD) of cacao, presents thirteen putative TLPs from which four are expressed during WBD progression. One of them is similar to small TLPs, which are present in phytopathogenic basidiomycete, such as wheat stem rust fungus Puccinia graminis. Fungi genomes annotation and phylogenetic data revealed a larger number of TLPs in basidiomycetes when comparing with ascomycetes, suggesting that these proteins could be involved in specific traits of mushroom-forming species. Based on the present data, we discuss the contribution of TLPs in the combat against fungal competitors and hypothesize a role of these proteins in M. perniciosa pathogenicity. PMID:26367180

  20. Rhynchostomatoid fungi occurring on Proteaceae.

    PubMed

    Lee, Seonju; Groenewald, J Z Ewald; Taylor, Joanne E; Roets, Francois; Crous, Pedro W

    2003-01-01

    A new ascomycete fungus, with long-necked perithecia having central ostioles and striate ascospores, was isolated from flowerheads of Protea burchellii and P. laurifolia in South Africa and is described here as Rhynchostoma proteae sp. nov. Sequence data obtained from the small-subunit ribosomal DNA (SSU nrDNA) place this fungus with 100% bootstrap support in a clade containing the type species of Rhynchostoma, R. minutum. A similar fungus with verruculose ascospores also was observed on a member of the Proteaceae from Australia, Lomatia polymorpha, which is described here as Rhynchomeliola lomatiae sp. nov. These two species are illustrated and contrasted with a third species from Proteaceae, Rhynchomeliola australiense, known from Grevillea in Australia. PMID:21148997

  1. The P450 Monooxygenase BcABA1 Is Essential for Abscisic Acid Biosynthesis in Botrytis cinerea

    PubMed Central

    Siewers, Verena; Smedsgaard, Jrn; Tudzynski, Paul

    2004-01-01

    The phytopathogenic ascomycete Botrytis cinerea is known to produce abscisic acid (ABA), which is thought to be involved in host-pathogen interaction. Biochemical analyses had previously shown that, in contrast to higher plants, the fungal ABA biosynthesis probably does not proceed via carotenoids but involves direct cyclization of farnesyl diphosphate and subsequent oxidation steps. We present here evidence that this direct pathway is indeed the only one used by an ABA-overproducing strain of B. cinerea. Targeted inactivation of the gene bccpr1 encoding a cytochrome P450 oxidoreductase reduced the ABA production significantly, proving the involvement of P450 monooxygenases in the pathway. Expression analysis of 28 different putative P450 monooxygenase genes revealed two that were induced under ABA biosynthesis conditions. Targeted inactivation showed that one of these, bcaba1, is essential for ABA biosynthesis: ?Bcaba1 mutants contained no residual ABA. Thus, bcaba1 represents the first identified fungal ABA biosynthetic gene. PMID:15240257

  2. The chestnut blight fungus for studies on virus/host and virus/virus interactions: from a natural to a model host.

    PubMed

    Eusebio-Cope, Ana; Sun, Liying; Tanaka, Toru; Chiba, Sotaro; Kasahara, Shin; Suzuki, Nobuhiro

    2015-03-01

    The chestnut blight fungus, Cryphonectria parasitica, is an important plant pathogenic ascomycete. The fungus hosts a wide range of viruses and now has been established as a model filamentous fungus for studying virus/host and virus/virus interactions. This is based on the development of methods for artificial virus introduction and elimination, host genome manipulability, available host genome sequence with annotations, host mutant strains, and molecular tools. Molecular tools include sub-cellular distribution markers, gene expression reporters, and vectors with regulatable promoters that have been long available for unicellular organisms, cultured cells, individuals of animals and plants, and certain filamentous fungi. A comparison with other filamentous fungi such as Neurospora crassa has been made to establish clear advantages and disadvantages of C. parasitica as a virus host. In addition, a few recent studies on RNA silencing vs. viruses in this fungus are introduced. PMID:25454384

  3. Transformation of Botrytis cinerea by direct hyphal blasting or by wound-mediated transformation of sclerotia

    PubMed Central

    2011-01-01

    Background Botrytis cinerea is a haploid necrotrophic ascomycete which is responsible for 'grey mold' disease in more than 200 plant species. Broad molecular research has been conducted on this pathogen in recent years, resulting in the sequencing of two strains, which has generated a wealth of information toward developing additional tools for molecular transcriptome, proteome and secretome investigations. Nonetheless, transformation protocols have remained a significant bottleneck for this pathogen, hindering functional analysis research in many labs. Results In this study, we tested three different transformation methods for B. cinerea: electroporation, air-pressure-mediated and sclerotium-mediated transformation. We demonstrate successful transformation with three different DNA constructs using both air-pressure- and sclerotium-mediated transformation. Conclusions These transformation methods, which are fast, simple and reproducible, can expedite functional gene analysis of B. cinerea. PMID:22188865

  4. Fusarium circinatum isolates from northern Spain are commonly infected by three distinct mitoviruses.

    PubMed

    Vainio, Eeva J; Martínez-Álvarez, Pablo; Bezos, Diana; Hantula, Jarkko; Diez, Julio J

    2015-08-01

    Pitch canker is a serious disease of pines caused by the ascomycete fungus Gibberella circinata (anamorph = Fusarium circinatum). Three distinct mitovirus strains have been described in this fungus: Fusarium circinatum mitovirus 1 (FcMV1), FcMV2-1 and FcMV2-2. Here, we investigated the frequency and population variation of these viruses and closely related sequence variants in northern Spain using RT-PCR and sequencing. Each virus strain and similar sequence variants shared >95 % sequence identity and were collectively designated as virus types. All virus types were relatively common in Spain, with estimated prevalence of 18.5 %, 8.9 % and 16.3 % for FcMV1, FcMV2-1 and FcMV2-2, respectively. PMID:26025157

  5. Social wasps are a Saccharomyces mating nest

    PubMed Central

    Stefanini, Irene; Dapporto, Leonardo; Berná, Luisa; Polsinelli, Mario; Turillazzi, Stefano; Cavalieri, Duccio

    2016-01-01

    The reproductive ecology of Saccharomyces cerevisiae is still largely unknown. Recent evidence of interspecific hybridization, high levels of strain heterozygosity, and prion transmission suggest that outbreeding occurs frequently in yeasts. Nevertheless, the place where yeasts mate and recombine in the wild has not been identified. We found that the intestine of social wasps hosts highly outbred S. cerevisiae strains as well as a rare S. cerevisiae×S. paradoxus hybrid. We show that the intestine of Polistes dominula social wasps favors the mating of S. cerevisiae strains among themselves and with S. paradoxus cells by providing a succession of environmental conditions prompting cell sporulation and spores germination. In addition, we prove that heterospecific mating is the only option for European S. paradoxus strains to survive in the gut. Taken together, these findings unveil the best hidden secret of yeast ecology, introducing the insect gut as an environmental alcove in which crosses occur, maintaining and generating the diversity of the ascomycetes. PMID:26787874

  6. Phylogeny of nematode-trapping fungi based on 18S rDNA sequences.

    PubMed

    Ahrén, D; Ursing, B M; Tunlid, A

    1998-01-15

    The small subunit (SSU) ribosomal DNA (18S rDNA) from 15 species of nematode-trapping fungi and closely related non-parasitic species were sequenced. Phylogenetic analysis indicated that species within the genera of Arthrobotrys, Dactylaria, Dactylella, Monacrosporium and Duddingtonia formed a monophyletic and isolated clade among an unresolved cluster of apothecial ascomycetes. The phylogenetic patterns within this clade were not concordant with the morphology of the conidia nor the conidiophores, but rather with that of the infection structures. The results from the different methods of tree reconstruction supported three lineages; the species having constricting rings, the non-parasitic species and the species having various adhesive structures (nets, hyphae, knobs and non-constricting rings) to infect nematodes. PMID:9465391

  7. Sympatric genetic differentiation of a generalist pathogenic fungus, Botrytis cinerea, on two different host plants, grapevine and bramble.

    PubMed

    Fournier, E; Giraud, T

    2008-01-01

    Prime candidates for sympatric ecological divergence include parasites that differentiate via host shifts, because different host species exert strong disruptive selection and because both hosts and parasites are continually co-evolving. Sympatric divergence may be fostered even more strongly in phytopathogenic fungi, in particular those where sex must occur on the host, which allows adaptation alone to restrict gene flow between populations developing on different hosts. We sampled populations of Botrytis cinerea, a generalist ascomycete fungus, on sympatric grapes and brambles in six regions in France. Microsatellite data were analyzed using standard population genetics, a population graph analysis and a Bayesian approach. In addition to confirming that B. cinerea reproduces sexually, our results showed that the fungal populations on the two hosts were significantly differentiated, indicating restricted gene flow, even in sympatry. In contrast, only weak geographical differentiation could be detected. These results support the possibility of sympatric divergence associated with host use in generalist parasites. PMID:18028352

  8. Protein kinase A signaling and calcium ions are major players in PAF mediated toxicity against Aspergillus niger.

    PubMed

    Binder, Ulrike; Benčina, Mojca; Fizil, Ádám; Batta, Gyula; Chhillar, Anil K; Marx, Florentine

    2015-05-01

    The Penicillium chrysogenum antifungal protein PAF is toxic against potentially pathogenic Ascomycetes. We used the highly sensitive aequorin-expressing model Aspergillus niger to identify a defined change in cytoplasmic free Ca(2+) dynamics in response to PAF. This Ca(2+) signature depended on an intact positively charged lysine-rich PAF motif. By combining Ca(2+) measurements in A. niger mutants with deregulated cAMP/protein kinase A (PKA) signaling, we proved the interconnection of Ca(2+) perturbation and cAMP/PKA signaling in the mechanistic function of PAF. A deep understanding of the mode of action of PAF is an invaluable prerequisite for its future application as new antifungal drug. PMID:25882631

  9. Social wasps are a Saccharomyces mating nest.

    PubMed

    Stefanini, Irene; Dapporto, Leonardo; Berná, Luisa; Polsinelli, Mario; Turillazzi, Stefano; Cavalieri, Duccio

    2016-02-23

    The reproductive ecology of Saccharomyces cerevisiae is still largely unknown. Recent evidence of interspecific hybridization, high levels of strain heterozygosity, and prion transmission suggest that outbreeding occurs frequently in yeasts. Nevertheless, the place where yeasts mate and recombine in the wild has not been identified. We found that the intestine of social wasps hosts highly outbred S. cerevisiae strains as well as a rare S. cerevisiae×S. paradoxus hybrid. We show that the intestine of Polistes dominula social wasps favors the mating of S. cerevisiae strains among themselves and with S. paradoxus cells by providing a succession of environmental conditions prompting cell sporulation and spores germination. In addition, we prove that heterospecific mating is the only option for European S. paradoxus strains to survive in the gut. Taken together, these findings unveil the best hidden secret of yeast ecology, introducing the insect gut as an environmental alcove in which crosses occur, maintaining and generating the diversity of the ascomycetes. PMID:26787874

  10. Variation of Soil Mycoflora in Decomposition of Rice Stubble from Rice-wheat Cropping System

    PubMed Central

    Sinha, Asha

    2007-01-01

    The colonization pattern and extent of decay produced in paddy stubble by soil inhabiting mycoflora were done by using nylon net bag technique. Among the three methods used for isolation of fungi, dilution plate technique recorded the highest number of fungi followed by damp chamber and direct observation method. Nutrient availability and climatic conditions (temperature, humidity and rainfall) influenced the occurrence and colonization pattern of fungi. Maximum fungal population was recorded in October (48.99 × 104/g dry litter) and minimum in May (11.41 × 104/g dry litter). Distribution of Deuteromycetous fungi was more in comparison to Zygomycetes, oomycetes and ascomycetes. In the early stage of decomposition Mucor racemosus, Rhizopus nigricans, Chaetomium globosum and Gliocladium species were found primarly whereas at later stages of decomposition preponderance of Aspergillus candidus, Torula graminis, Cladosporiun cladosporioides and Aspergillus luchuensis was recorded. PMID:24015096

  11. Variation of Soil Mycoflora in Decomposition of Rice Stubble from Rice-wheat Cropping System.

    PubMed

    Vibha; Sinha, Asha

    2007-12-01

    The colonization pattern and extent of decay produced in paddy stubble by soil inhabiting mycoflora were done by using nylon net bag technique. Among the three methods used for isolation of fungi, dilution plate technique recorded the highest number of fungi followed by damp chamber and direct observation method. Nutrient availability and climatic conditions (temperature, humidity and rainfall) influenced the occurrence and colonization pattern of fungi. Maximum fungal population was recorded in October (48.99 × 10(4)/g dry litter) and minimum in May (11.41 × 10(4)/g dry litter). Distribution of Deuteromycetous fungi was more in comparison to Zygomycetes, oomycetes and ascomycetes. In the early stage of decomposition Mucor racemosus, Rhizopus nigricans, Chaetomium globosum and Gliocladium species were found primarly whereas at later stages of decomposition preponderance of Aspergillus candidus, Torula graminis, Cladosporiun cladosporioides and Aspergillus luchuensis was recorded. PMID:24015096

  12. On the trail of a cereal killer: recent advances in Fusarium graminearum pathogenomics and host resistance.

    PubMed

    Kazan, Kemal; Gardiner, Donald M; Manners, John M

    2012-05-01

    The ascomycete fungal pathogen Fusarium graminearum (sexual stage: Gibberella zeae) causes the devastating head blight or scab disease on wheat and barley, and cob or ear rot disease on maize. Fusarium graminearum infection causes significant crop and quality losses. In addition to roles as virulence factors during pathogenesis, trichothecene mycotoxins (e.g. deoxynivalenol) produced by this pathogen constitute a significant threat to human and animal health if consumed in respective food or feed products. In the last few years, significant progress has been made towards a better understanding of the processes involved in F. graminearum pathogenesis, toxin biosynthesis and host resistance mechanisms through the use of high-throughput genomic and phenomic technologies. In this article, we briefly review these new advances and also discuss how future research can contribute to the development of sustainable plant protection strategies against this important plant pathogen. PMID:22098555

  13. On the trail of a cereal killer: recent advances in Fusarium graminearum pathogenomics and host resistance.

    TOXLINE Toxicology Bibliographic Information

    Kazan K; Gardiner DM; Manners JM

    2012-05-01

    The ascomycete fungal pathogen Fusarium graminearum (sexual stage: Gibberella zeae) causes the devastating head blight or scab disease on wheat and barley, and cob or ear rot disease on maize. Fusarium graminearum infection causes significant crop and quality losses. In addition to roles as virulence factors during pathogenesis, trichothecene mycotoxins (e.g. deoxynivalenol) produced by this pathogen constitute a significant threat to human and animal health if consumed in respective food or feed products. In the last few years, significant progress has been made towards a better understanding of the processes involved in F. graminearum pathogenesis, toxin biosynthesis and host resistance mechanisms through the use of high-throughput genomic and phenomic technologies. In this article, we briefly review these new advances and also discuss how future research can contribute to the development of sustainable plant protection strategies against this important plant pathogen.

  14. Scytalidium and scytalidiosis: what's new in 2012?

    PubMed

    Machouart, M; Menir, P; Helenon, R; Quist, D; Desbois, N

    2013-03-01

    Fungi belonging to the genus Scytalidium are widespread around the world. Among them, two species are responsible for human superficial infections mimicking dermatophytosis: Neoscytalidium dimidiatum and Scytalidium hyalinum. Whereas these ascomycetous fungi are endemic in tropical or subtropical countries, both species have a different geographical distribution. Scytalidiosis represents approximately 40% of dermatomycoses in these areas. A few cases of invasive infections due to Scytalidium sp. have also been reported, assessing the ability of these fungi to behave as opportunists. Here we have reviewed the data on N. dimidiatum and S. hyalinum concerning their classification, clinical features, diagnosis and treatment. We also have presented the example of a specific consultation dedicated to nails in Martinique, in order to optimize the diagnosis and treatment of onychomycosis, many of which being due to Scytalidium sp. Even if Scytalidium cases are still rare in temperate countries, imported cases may increase in the future due to immigration and travel. PMID:23416171

  15. Phylogenetic and biological species diversity within the Neurospora tetrasperma complex.

    PubMed

    Menkis, A; Bastiaans, E; Jacobson, D J; Johannesson, H

    2009-09-01

    The objective of this study was to explore the evolutionary history of the morphologically recognized filamentous ascomycete Neurospora tetrasperma, and to reveal the genetic and reproductive relationships among its individuals and populations. We applied both phylogenetic and biological species recognition to a collection of strains representing the geographic and genetic diversity of N. tetrasperma. First, we were able to confirm a monophyletic origin of N. tetrasperma. Furthermore, we found nine phylogenetic species within the morphospecies. When using the traditional broad biological species recognition all investigated strains of N. tetrasperma constituted a single biological species. In contrast, when using a quantitative measurement of the reproductive success, incorporating characters such as viability and fertility of offspring, we found a high congruence between the phylogenetic and biological species recognition. Taken together, phylogenetically and biologically defined groups of individuals exist in N. tetrasperma, and these should be taken into account in future studies of its life history traits. PMID:19682307

  16. The importance of molecular analyses for understanding the genetic diversity of Histoplasma capsulatum: an overview.

    PubMed

    Vite-Garín, Tania; Estrada-Bárcenas, Daniel Alfonso; Cifuentes, Joaquín; Taylor, Maria Lucia

    2014-01-01

    Advances in the classification of the human pathogen Histoplasma capsulatum (H. capsulatum) (ascomycete) are sustained by the results of several genetic analyses that support the high diversity of this dimorphic fungus. The present mini-review highlights the great genetic plasticity of H. capsulatum. Important records with different molecular tools, mainly single- or multi-locus sequence analyses developed with this fungus, are discussed. Recent phylogenetic data with a multi-locus sequence analysis using 5 polymorphic loci support a new clade and/or phylogenetic species of H. capsulatum for the Americas, which was associated with fungal isolates obtained from the migratory bat Tadarida brasiliensis. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012). PMID:24252830

  17. Comprehensive Metabolomic, Lipidomic and Microscopic Profiling of Yarrowia lipolytica during Lipid Accumulation Identifies Targets for Increased Lipogenesis

    PubMed Central

    Pomraning, Kyle R.; Wei, Siwei; Karagiosis, Sue A.; Kim, Young-Mo; Dohnalkova, Alice C.; Arey, Bruce W.; Bredeweg, Erin L.; Orr, Galya; Metz, Thomas O.; Baker, Scott E.

    2015-01-01

    Yarrowia lipolytica is an oleaginous ascomycete yeast that accumulates large amounts of lipids and has potential as a biofuel producing organism. Despite a growing scientific literature focused on lipid production by Y. lipolytica, there remain significant knowledge gaps regarding the key biological processes involved. We applied a combination of metabolomic and lipidomic profiling approaches as well as microscopic techniques to identify and characterize the key pathways involved in de novo lipid accumulation from glucose in batch cultured, wild-type Y. lipolytica. We found that lipids accumulated rapidly and peaked at 48 hours during the five day experiment, concurrent with a shift in amino acid metabolism. We also report that exhaustion of extracellular sugars coincided with thickening of the cell wall, suggesting that genes involved in cell wall biogenesis may be a useful target for improving the efficiency of lipid producing yeast strains. PMID:25905710

  18. Nutritional physiology and taxonomy of human-pathogenic Cladosporium-Xylohypha species.

    PubMed

    de Hoog, G S; Guho, E; Masclaux, F; Gerrits van den Ende, A H; Kwon-Chung, K J; McGinnis, M R

    1995-01-01

    Physiological profiles of type, authentic and some additional isolates of Cladosporium-Xylohypha species of purported herpotrichiellaceous relationship are established. This group comprises melanized catenate hyphomycetes which are prevalently found on the human host. The species are excluded from the genus Cladosporium and are classified in the genus Cladophialophora. Taeniolella boppii is also transferred to this genus. Cladosporium bantianum (= Xylohypha emmonsii) and C. trichoides are considered conspecific and are now referred to as Cladophialophora bantiana. Meso-erythritol, L-arabinitol, ethanol and growth at 40 degrees C are found to be the most useful criteria for species distinction. The species Cladosporium carrionii is found to be heterogeneous. The anamorph of the saprophytic ascomycete Capronia pilosella is morphologically similar to an authentic strain of Cladosporium carrionii, but physiologically distinct. A diagnostic key for the recognized Cladophialophora species and to morphologically similar taxa is provided. PMID:8544087

  19. Characterization of three DNA transposons in the Dutch elm disease fungi and evidence of repeat-induced point (RIP) mutations.

    PubMed

    Bouvet, Guillaume F; Jacobi, Volker; Bernier, Louis

    2007-05-01

    Transposable elements (TEs) are fundamental components of eukaryotic genomes and can contribute in various ways to genome plasticity and evolution. We describe here the first three DNA transposons in the Dutch elm disease (DED) pathogens Ophiostoma ulmi and O. novo-ulmi, named OPHIO1, OPHIO2 and OPHIO3. We demonstrate that OPHIO transposons, which show high homology to Fot1/pogo TEs within the Tc1/mariner superfamily, have different distribution patterns and specificity in the DED fungi and that interspecific hybrids could act as genetic bridges for transmission of TEs between closely related fungal species. OPHIO3 was found to have undergone repeat-induced point mutations (RIP). We have also developed a complementary method to Margolin's ratios based on the computation of cumulative transition scores (CTS) in order to visualize rapidly RIP signatures on individual DNA strands of OPHIO transposons and TEs found in other ascomycete fungi. PMID:17291794

  20. Development of a Selective Medium for the Fungal Pathogen Fusarium graminearum Using Toxoflavin Produced by the Bacterial Pathogen Burkholderia glumae

    PubMed Central

    Jung, Boknam; Lee, Sehee; Ha, Jiran; Park, Jong-Chul; Han, Sung-Sook; Hwang, Ingyu; Lee, Yin-Won; Lee, Jungkwan

    2013-01-01

    The ascomycete fungus Fusarium graminearum is a major causal agent for Fusarium head blight in cereals and produces mycotoxins such as trichothecenes and zearalenone. Isolation of the fungal strains from air or cereals can be hampered by various other airborne fungal pathogens and saprophytic fungi. In this study, we developed a selective medium specific to F. graminearum using toxoflavin produced by the bacterial pathogen Burkholderia glumae. F. graminearum was resistant to toxoflavin, while other fungi were sensitive to this toxin. Supplementing toxoflavin into medium enhanced the isolation of F. graminearum from rice grains by suppressing the growth of saprophytic fungal species. In addition, a medium with or without toxoflavin exposed to wheat fields for 1 h had 84% or 25%, respectively, of colonies identified as F. graminearum. This selection medium provides an efficient tool for isolating F. graminearum, and can be adopted by research groups working on genetics and disease forecasting. PMID:25288974