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Sample records for huitlacoche ustilago maydis

  1. Selection of aggressive pathogenic and solopathogenic strains of Ustilago maydis to improve Huitlacoche production

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    Porfirio Raúl Galicia-García

    Full Text Available ABSTRACT Ustilago maydis is a basidiomycete known as the causative agent of 'common smut', worldwide disease of maize that is recognized by the galls it forms, which have considerable potential as a gourmet food. Results of infection are quite variable, even under optimal greenhouse conditions. In order to find pathogenic strains able to be used as a highly infective and stable inoculum for the successful production of galls either in greenhouses or in the field, ears with gall symptoms containing teliospores were recovered from maize plants. The teliospores were suspended in water and plated on nutrient-rich medium. Twenty-six colonies developed, containing three types of yeast-like colonies: saprotrophic, pathogenic, and solopathogenic. DAPI staining confirmed the presence of solopathogenic strains with diploid sporidia. Groups of different mating types were found when pairs of the 26 strains were arranged resembling partial-diallel combinations. Amplification of the partial b locus revealed that the strains found harbor the alleles b3 and b4, allowing the formation in dikaryotic strains of heterodimeric regulatory proteins associated with fungal development and pathogenicity. In this study, we isolated compatible haploid and solopathogenic diploid strains for their high capacity for inducing smut.

  2. Huitlacoche (corn smut), caused by the phytopathogenic fungus Ustilago maydis, as a functional food.

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    Juárez-Montiel, Margarita; Ruiloba de León, Sandra; Chávez-Camarillo, Griselda; Hernández-Rodríguez, César; Villa-Tanaca, Lourdes

    2011-01-01

    In recent years the need has arisen to study and develop (or re-discover) foods that have nutritional characteristics as well as specific functions, such as improving health and/or reducing the risk of disease. For this reason knowledge of the nutritional value of food is important to promote greater consumer acceptance. In Mexico huitlacoche (also, cuitlacoche) has traditionally been prized as a delicacy since the time of the Aztecs and is currently being studied as a potential functional food and as a producer of natural bioactive substances that are used in fortifying foods. To present an updated review about the properties of the huitlacoche (corn smut) as functional food. A bibliographic search was performed and data were discussed. The data of the works reviewed here show that huitlacoche contains many compounds that confer to it unique organoleptic and nutraceutical characteristics. The content of bioactive substances in huitlacoche supports the proposal that this is a good functional food as well as producer of compounds to enrich other foods. Copyright © 2010 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  3. Effect of huitlacoche (Ustilago maydis DC Corda paste addition on functional, chemical and textural properties of tortilla chips

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    Karla Yuritzi Amador-Rodríguez

    2015-09-01

    Full Text Available AbstractThis study analyzed the addition of huitlacoche paste (HP in baked tortilla chips (TC, evaluating its effects on functional, physicochemical and structural changes during processing. Two blue corn grains were nixtamalized, stone milled, air dried and milled to obtain flour; commercial blue corn flour (TM1 and commercial TC (TM2 were used as controls. Additions of 0, 3, 6 and 9% of HP were formulated; masas were prepared at 55% moisture content (MC, precooked and baked in an industrial machine. TC crispiness was influenced by grain characteristics and percentage of HP. Huitlacoche paste addition caused an increase in total dietary fiber (from 5.27 to 14.54%, total soluble phenolics content (from 17.52 to 37.60 mg GAE/100 g and antioxidant capacity (from 6.74 to 7.98 μmol TE/g in TC. Results suggest that tortilla chips added with huitlacoche can be an alternative to prepare this traditional edible fungus and produce healthier snacks, not fried and enriched with bioactive compounds.

  4. Cytotoxicity of Ustilago maydis isolated from maize

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    Magdalena Twarużek

    2013-03-01

    Full Text Available The main pathogen of maize are fungi of the genus Fusarium. Besides phytopathogenic Fusarium, Ustilago maydis is another fungal genus affecting maize yields, causing lesions, known as smut. The objective of the study was evaluation of the cytotoxicity of Ustilago maydis isolated from maize. Nine Ustilago maydis strains were selected to a detailed evaluation of their cytotoxicity using a 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT test. Ustilago maydis strains showed medium and high cytotoxicity compared to control. High levels of cytotoxicity of Ustilago maydis may be indicative of their toxigenic potential.

  5. Bioinformatic identification of Ustilago maydis meiosis genes.

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    Donaldson, Michael E; Saville, Barry J

    2008-08-01

    In the corn smut pathogen, Ustilago maydis, meiosis and teliospore germination are temporally linked. We review teliospore dormancy and germination in U. maydis and present an overview of meiosis in basidiomycetes. The relevant available expressed sequence tag data is discussed, the databases used in reciprocal best hit blastp analysis are presented and potential U. maydis meiosis genes are identified. The implications of identifying these genes are discussed and hypotheses are presented regarding the control of meiosis in U. maydis.

  6. Initiation of meiotic recombination in Ustilago maydis

    National Research Council Canada - National Science Library

    Kojic, Milorad; Sutherland, Jeanette H; Pérez-Martín, José; Holloman, William K

    2013-01-01

    .... Ustilago maydis, a biotrophic fungus that parasitizes maize, has long been utilized as an experimental system for studying recombination, but it has not been clear when in the life cycle meiotic recombination initiates. U...

  7. The homologous recombination system of Ustilago maydis

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    Holloman, William K.; Schirawski, Jan; Holliday, Robin

    2008-01-01

    Homologous recombination is a high fidelity, template-dependent process that is used in repair of damaged DNA, recovery of broken replication forks, and disjunction of homologous chromosomes in meiosis. Much of what is known about recombination genes and mechanisms comes from studies on baker's yeast. Ustilago maydis, a basidiomycete fungus, is distant evolutionarily from baker's yeast and so offers the possibility of gaining insight into recombination from an alternative perspective. Here we...

  8. The role of repellents and hydrophobins in Ustilago maydis

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    Teertstra, W.R.

    2009-01-01

    Ustilago maydis is an important model organism to study fungal pathogenicity. U. maydis can grow yeast-like and filamentous. In the latter form this fungus infects maize. In my Thesis the expression and function of hydrophobins and repellents of U. maydis were studied. Hydrophobins are produced by f

  9. The role of repellents and hydrophobins in Ustilago maydis

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    Teertstra, W.R.

    2009-01-01

    Ustilago maydis is an important model organism to study fungal pathogenicity. U. maydis can grow yeast-like and filamentous. In the latter form this fungus infects maize. In my Thesis the expression and function of hydrophobins and repellents of U. maydis were studied. Hydrophobins are produced by f

  10. The homologous recombination system of Ustilago maydis.

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    Holloman, William K; Schirawski, Jan; Holliday, Robin

    2008-08-01

    Homologous recombination is a high fidelity, template-dependent process that is used in repair of damaged DNA, recovery of broken replication forks, and disjunction of homologous chromosomes in meiosis. Much of what is known about recombination genes and mechanisms comes from studies on baker's yeast. Ustilago maydis, a basidiomycete fungus, is distant evolutionarily from baker's yeast and so offers the possibility of gaining insight into recombination from an alternative perspective. Here we have surveyed the genome of U. maydis to determine the composition of its homologous recombination system. Compared to baker's yeast, there are fundamental differences in the function as well as in the repertoire of dedicated components. These include the use of a BRCA2 homolog and its modifier Dss1 rather than Rad52 as a mediator of Rad51, the presence of only a single Rad51 paralog, and the absence of Dmc1 and auxiliary meiotic proteins.

  11. Initiation of meiotic recombination in Ustilago maydis.

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    Kojic, Milorad; Sutherland, Jeanette H; Pérez-Martín, José; Holloman, William K

    2013-12-01

    A central feature of meiosis is the pairing and recombination of homologous chromosomes. Ustilago maydis, a biotrophic fungus that parasitizes maize, has long been utilized as an experimental system for studying recombination, but it has not been clear when in the life cycle meiotic recombination initiates. U. maydis forms dormant diploid teliospores as the end product of the infection process. Upon germination, teliospores complete meiosis to produce four haploid basidiospores. Here we asked whether the meiotic process begins when teliospores germinate or at an earlier stage in development. When teliospores homozygous for a cdc45 mutation temperature sensitive for DNA synthesis were germinated at the restrictive temperature, four nuclei became visible. This implies that teliospores have already undergone premeiotic DNA synthesis and suggests that meiotic recombination initiates at a stage of infection before teliospores mature. Determination of homologous recombination in plant tissue infected with U. maydis strains heteroallelic for the nar1 gene revealed that Nar(+) recombinants were produced at a stage before teliospore maturation. Teliospores obtained from a spo11Δ cross were still able to germinate but the process was highly disturbed and the meiotic products were imbalanced in chromosomal complement. These results show that in U. maydis, homologous recombination initiates during the infection process and that meiosis can proceed even in the absence of Spo11, but with loss of genomic integrity.

  12. Plant surface cues prime Ustilago maydis for biotrophic development

    National Research Council Canada - National Science Library

    Lanver, Daniel; Berndt, Patrick; Tollot, Marie; Naik, Vikram; Vranes, Miroslav; Warmann, Tobias; Münch, Karin; Rössel, Nicole; Kahmann, Regine

    2014-01-01

    .... To unravel the program inducing filaments and appressoria in the biotrophic smut fungus Ustilago maydis, we exposed cells to a hydrophobic surface and the cutin monomer 16-hydroxy hexadecanoic acid...

  13. Genetics of morphogenesis and pathogenic development of Ustilago maydis.

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    Ustilago maydis has emerged as an important model system for the study of fungi. Like many fungi, U. maydis undergoes remarkable morphological transitions throughout its lifecycle. Fusion of compatible, budding, haploid cells leads to the production of a filamentous dikaryon that penetrates and colo...

  14. Ustilago maydis: ascenso de un hongo mexicano de la gastronomía local al mundo científico

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    José Ruiz Herrera

    2008-01-01

    En el presente trabajo se describe la naturaleza del hongo Ustilago maydis, su papel como causante de una enfermedad del maíz, su uso en la cocina mexicana y las condiciones que lo llevaron a convertirse en material de estudio científico, además de algunas características descollantes de su fisiología, genética y bioquímica. U. maydis es un hongo patógeno biotrófico específico del maíz, y el teozintle, siendo el agente causal del "huitlacoche" o carbón común, una enfermedad de distribución m...

  15. RNA polymerase activity of Ustilago maydis virus

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    Yie, S.W.

    1986-01-01

    Ustilago maydis virus has an RNA polymerase enzyme which is associated with virion capsids. In the presence of Mg/sup 2 +/ ion and ribonucleotide triphosphate, the enzyme catalyzes the in vitro synthesis of mRNA by using dsRNA as a template. The products of the UmV RNA polymerase were both ssRNA and dsRNA. The dsRNA was determined by characteristic mobilities in gel electrophoresis, lack of sensitivity to RNase, and specific hybridization tests. The ssRNAs were identified by elution from a CF-11 column and by their RNase sensitivity. On the basis of the size of ssRNAs, it was concluded that partial transcripts were produced from H dsRNA segments, and full length transcripts were produced from M and L dsRNA segments. The following observations indicates that transcription occurs by strand displacement; (1) Only the positive strand of M2 dsRNA was labeled by the in vitro reaction. (2) The M2 dsRNA which had been labeled with /sup 32/''P-UTP in vitro could be chased from dsRNA with unlabeled UTP. The transcription products of three UmV strains were compared, and the overall pattern of transcription was very similar among them.

  16. Identificación de las proteínas secretadas por el hongo Ustilago maydis (DeCandole) Corda (Basidiomiceto) cultivado en condiciones in vitro

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    Andrés Adolfo Estrada-Luna; Alicia Chagolla López; Hilda Eréndira Ramos Aboites; Angelina Guerrero Ambriz; José Ruiz Herrera

    2010-01-01

    Introducción: Ustilago maydis es un hongo basidiomiceto que infecta al maíz y teozintle produciendo una enfermedad conocida como carbón común o huitlacoche. Actualmente no existen reportes acerca del secretoma del hongo cultivado bajo condiciones in vitro. Un estudio de esta naturaleza permitiría caracterizar los genes involucrados en varios procesos importantes, entre los que se tienen aquellos relacionados con la nutrición, la patogenicidad y la diferenciación del hongo. El objetivo de esta...

  17. Establishment of compatibility in the Ustilago maydis/maize pathosystem

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    Doehlemann, Gunther; Wahl, Ramon; Vranes, Miroslav; de Vries, Ronald P; Kämper, Jörg; Kahmann, Regine

    2008-01-01

    The fungus Ustilago maydis is a biotrophic pathogen parasitizing on maize. The most prominent symptoms of the disease are large tumors in which fungal proliferation and spore differentiation occur. In this study, we have analyzed early and late tumor stages by confocal microscopy. We show that funga

  18. The virally encoded killer proteins from Ustilago maydis

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    Several strains of Ustilago maydis, a causal agent of corn smut disease, exhibit a 'killer' phenotype that is due to persistent infection by double-stranded RNA Totiviruses. These viruses produce potent killer proteins that are secreted by the host. This is a rare example of virus/host symbiosis in ...

  19. Disruption of an ADE6 Homolog of Ustilago maydis

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    Ustilago maydis secretes iron-binding compounds during times of iron depletion. A putative homolog of the Sacharromyces cereviseae ADE6 and Escherichia coli purL genes was identified near a multigenic complex, which contains two genes sid1 and sid2 involved in a siderophore biosynthetic pathway. The...

  20. Establishment of compatibility in the Ustilago maydis/maize pathosystem

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    Doehlemann, Gunther; Wahl, Ramon; Vranes, Miroslav; de Vries, Ronald P; Kämper, Jörg; Kahmann, Regine

    2008-01-01

    The fungus Ustilago maydis is a biotrophic pathogen parasitizing on maize. The most prominent symptoms of the disease are large tumors in which fungal proliferation and spore differentiation occur. In this study, we have analyzed early and late tumor stages by confocal microscopy. We show that funga

  1. Towards understanding the extreme radiation resistance of Ustilago maydis.

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    Holloman, William K; Schirawski, Jan; Holliday, Robin

    2007-12-01

    Ustilago maydis is a phytopathogenic fungus exhibiting extreme resistance to UV and ionizing radiation. The molecular mechanisms underlying this resistance are as yet unknown. The recently determined genome sequence was examined for clues to the radiation resistance, focusing on proteins in homologous recombination, but there was little that was unusual about them. Furthermore, by comparison, its recombinational repair system seems to be only minimally related to the extended synthesis-dependent DNA strand-annealing system of Deinococcus radiodurans. Thus, consideration should be given to the possibility that incremental structural changes in repair proteins or their elevated expression are the basis for the extreme radiation resistance in U. maydis. Evolution of a system enabling the survival of U. maydis under such conditions could be a secondary consequence of adaptation to an environment of continual genotoxic stress encountered in its habitat.

  2. Endocytosis in the plant-pathogenic fungus Ustilago maydis.

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    Fuchs, U; Steinberg, G

    2005-10-01

    Filamentous fungi are an important group of tip-growing organisms, which include numerous plant pathogens such as Magnaporthe grisea and Ustilago maydis. Despite their ecological and economical relevance, we are just beginning to unravel the importance of endocytosis in filamentous fungi. Most evidence for endocytosis in filamentous fungi is based on the use of endocytic tracer dyes that are taken up into the cell and delivered to the vacuole. Moreover, genomewide screening for candidate genes in Neurospora crassa and U. maydis confirmed the presence of most components of the endocytic machinery, indicating that endocytosis participates in filamentous growth. Indeed, it was shown that in U. maydis early endosomes cluster at sites of growth, where they support morphogenesis and polar growth, most likely via endosome-based membrane recycling. In humans, such recycling processes to the plasma membrane involve small GTPases such as Rab4. A homologue of this protein is encoded in the genome of U. maydis but is absent from the yeast Saccharomyces cerevisiae, suggesting that Rab4-mediated recycling is important for filamentous growth. Furthermore, human Rab4 regulates traffic of early endosomes along microtubules, and a similar microtubule-based transport is described for U. maydis. These observations suggest that Rab4-like GTPases might regulate endosome- and microtubule-based recycling during tip growth of filamentous fungi.

  3. A maize-specifically expressed gene cluster in Ustilago maydis.

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    Basse, Christoph W; Kolb, Sebastian; Kahmann, Regine

    2002-01-01

    The corn pathogen Ustilago maydis requires its host plant maize for development and completion of its sexual cycle. We have identified the fungal mig2-1 gene as being specifically expressed during this biotrophic stage. Intriguingly, mig2-1 is part of a gene cluster comprising five highly homologous and similarly regulated genes designated mig2-1 to mig2-5. Deletion analysis of the mig2-1 promoter provides evidence for negative and positive regulation. The predicted polypeptides of all five genes lack significant homologies to known genes but have characteristic N-terminal secretion sequences. The secretion signals of mig2-1 and mig2-5 were shown to be functional, and secretion of a full length Mig2-1-eGFP fusion protein to the extracellular space was demonstrated. The central domains of the Mig2 proteins are highly variable whereas the C-termini are strongly conserved and share a characteristic pattern of eight cysteine residues. The mig2 gene cluster was conserved in a wide collection of U. maydis strains. Interestingly, some U. maydis isolates from South America had lost the mig2-4 gene as a result of a homologous recombination event. Furthermore, the related Ustilago scitaminea strain, which is pathogenic on sugar cane, appears to lack the mig2 cluster. We describe a model of how the mig2 cluster might have evolved and discuss its possible role in governing host interaction.

  4. Sugar Substrates for l-Lysine Fermentation by Ustilago maydis

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    Sánchez-Marroquín, A.; Ledezma, M.; Carreño, R.

    1970-01-01

    The extracellular production of l-lysine in media with cane sugar, blackstrap molasses, or clarified sugar-cane juice by a previously obtained mutant of Ustilago maydis was studied. Enzymatically inverted clarified juice (medium J-3) gave 2.9 g of lysine per liter under the following conditions: inoculum, 5%; pH 5.8; temperature, 30 C; KLa in the fermentors, 0.41 mmoles of O2 per liter per min; fermentation time, 72 hr. The concentrate, obtained by direct evaporation and drying of the fermentation broth, could be used as a possible feed supplement because of its amino-acid and vitamin content. PMID:5485081

  5. Ustilago maydis reprograms cell proliferation in maize anthers.

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    Gao, Li; Kelliher, Timothy; Nguyen, Linda; Walbot, Virginia

    2013-09-01

    The basidiomycete Ustilago maydis is a ubiquitous pathogen of maize (Zea mays), one of the world's most important cereal crops. Infection by this smut fungus triggers tumor formation in aerial plant parts within which the fungus sporulates. Using confocal microscopy to track U. maydis infection on corn anthers for 7 days post-injection, we found that U. maydis is located on the epidermis during the first 2 days, and has reached all anther lobe cell types by 3 days post-injection. Fungal infection alters cell-fate specification events, cell division patterns, host cell expansion and host cell senescence, depending on the developmental stage and cell type. Fungal effects on tassel and plant growth were also quantified. Transcriptome profiling using a dual organism microarray identified thousands of anther genes affected by fungal infection at 3 days post-injection during the cell-fate specification and rapid cell proliferation phases of anther development. In total, 4147 (17%) of anther-expressed genes were altered by infection, 2018 fungal genes were expressed in anthers, and 206 fungal secretome genes may be anther-specific. The results confirm that U. maydis deploys distinct genes to cause disease in specific maize organs, and suggest mechanisms by which the host plant is manipulated to generate a tumor.

  6. Biomass pretreatment affects Ustilago maydis in producing itaconic acid

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    Klement Tobias

    2012-04-01

    Full Text Available Abstract Background In the last years, the biotechnological production of platform chemicals for fuel components has become a major focus of interest. Although ligno-cellulosic material is considered as suitable feedstock, the almost inevitable pretreatment of this recalcitrant material may interfere with the subsequent fermentation steps. In this study, the fungus Ustilago maydis was used to produce itaconic acid as platform chemical for the synthesis of potential biofuels such as 3-methyltetrahydrofuran. No studies, however, have investigated how pretreatment of ligno-cellulosic biomass precisely influences the subsequent fermentation by U. maydis. Thus, this current study aims to first characterize U. maydis in shake flasks and then to evaluate the influence of three exemplary pretreatment methods on the cultivation and itaconic acid production of this fungus. Cellulose enzymatically hydrolysed in seawater and salt-assisted organic-acid catalysed cellulose were investigated as substrates. Lastly, hydrolysed hemicellulose from fractionated beech wood was applied as substrate. Results U. maydis was characterized on shake flask level regarding its itaconic acid production on glucose. Nitrogen limitation was shown to be a crucial condition for the production of itaconic acid. For itaconic acid concentrations above 25 g/L, a significant product inhibition was observed. Performing experiments that simulated influences of possible pretreatment methods, U. maydis was only slightly affected by high osmolarities up to 3.5 osmol/L as well as of 0.1 M oxalic acid. The production of itaconic acid was achieved on pretreated cellulose in seawater and on the hydrolysed hemicellulosic fraction of pretreated beech wood. Conclusion The fungus U. maydis is a promising producer of itaconic acid, since it grows as single cells (yeast-like in submerged cultivations and it is extremely robust in high osmotic media and real seawater. Moreover, U. maydis can grow on

  7. Plant surface cues prime Ustilago maydis for biotrophic development.

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    Daniel Lanver

    2014-07-01

    Full Text Available Infection-related development of phytopathogenic fungi is initiated by sensing and responding to plant surface cues. This response can result in the formation of specialized infection structures, so-called appressoria. To unravel the program inducing filaments and appressoria in the biotrophic smut fungus Ustilago maydis, we exposed cells to a hydrophobic surface and the cutin monomer 16-hydroxy hexadecanoic acid. Genome-wide transcriptional profiling at the pre-penetration stage documented dramatic transcriptional changes in almost 20% of the genes. Comparisons with the U. maydis sho1 msb2 double mutant, lacking two putative sensors for plant surface cues, revealed that these plasma membrane receptors regulate a small subset of the surface cue-induced genes comprising mainly secreted proteins including potential plant cell wall degrading enzymes. Targeted gene deletion analysis ascribed a role to up-regulated GH51 and GH62 arabinofuranosidases during plant penetration. Among the sho1/msb2-dependently expressed genes were several secreted effectors that are essential for virulence. Our data also demonstrate specific effects on two transcription factors that redirect the transcriptional regulatory network towards appressorium formation and plant penetration. This shows that plant surface cues prime U. maydis for biotrophic development.

  8. Plant surface cues prime Ustilago maydis for biotrophic development.

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    Lanver, Daniel; Berndt, Patrick; Tollot, Marie; Naik, Vikram; Vranes, Miroslav; Warmann, Tobias; Münch, Karin; Rössel, Nicole; Kahmann, Regine

    2014-07-01

    Infection-related development of phytopathogenic fungi is initiated by sensing and responding to plant surface cues. This response can result in the formation of specialized infection structures, so-called appressoria. To unravel the program inducing filaments and appressoria in the biotrophic smut fungus Ustilago maydis, we exposed cells to a hydrophobic surface and the cutin monomer 16-hydroxy hexadecanoic acid. Genome-wide transcriptional profiling at the pre-penetration stage documented dramatic transcriptional changes in almost 20% of the genes. Comparisons with the U. maydis sho1 msb2 double mutant, lacking two putative sensors for plant surface cues, revealed that these plasma membrane receptors regulate a small subset of the surface cue-induced genes comprising mainly secreted proteins including potential plant cell wall degrading enzymes. Targeted gene deletion analysis ascribed a role to up-regulated GH51 and GH62 arabinofuranosidases during plant penetration. Among the sho1/msb2-dependently expressed genes were several secreted effectors that are essential for virulence. Our data also demonstrate specific effects on two transcription factors that redirect the transcriptional regulatory network towards appressorium formation and plant penetration. This shows that plant surface cues prime U. maydis for biotrophic development.

  9. Ustilago maydis secondary metabolism-from genomics to biochemistry.

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    Bölker, Michael; Basse, Christoph W; Schirawski, Jan

    2008-08-01

    The dimorphic phytopathogenic fungus Ustilago maydis encounters different environments during its life cycle. As free-living unicellular haploid cell, the fungus must compete with other microorganisms for space and nutrients. As a pathogen, it also has to withstand the defense reactions of its host plant corn and to subvert the plant metabolism for its own purposes. During these interactions small molecules produced by the fungus serve important functions in the communication with its host and other organisms. The genome sequence of U. maydis makes it possible to deduce the full inventory of enzymatic functions that are involved in the production of these secondary metabolites. Although the fungus is known to secrete interesting small molecules the genome contains surprisingly few genes involved in the biosynthesis of polyketides (PKS) and non-ribosomal peptide synthetases (NRPS). Additional genes predicted to be part of secondary metabolism are located in subtelomeric regions suggesting that they are subject to high genetic and genomic variation. Here we review the pathways for the production of extracellular glycolipids that serve as biosurfactants, iron-chelating siderophores, tryptophan-derived indole pigments and indole acetic acid, the elucidation of which has greatly profited from the availability of the U. maydis genome sequence.

  10. Mating and Progeny Isolation in The Corn Smut Fungus Ustilago maydis

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    The corn smut pathogen, Ustilago maydis (U. maydis) (DC.) Corda, is a semi-obligate plant pathogenic fungus in the phylum Basidiomycota (Alexopoulos, Mims and Blackwell, 1996). The fungus can be easily cultured in its haploid yeast phase on common laboratory media. However, to complete its sexual cy...

  11. Chitinases Are Essential for Cell Separation in Ustilago maydis.

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    Langner, Thorsten; Öztürk, Merve; Hartmann, Sarah; Cord-Landwehr, Stefan; Moerschbacher, Bruno; Walton, Jonathan D; Göhre, Vera

    2015-09-01

    Chitin is an essential component of the fungal cell wall, providing rigidity and stability. Its degradation is mediated by chitinases and supposedly ensures the dynamic plasticity of the cell wall during growth and morphogenesis. Hence, chitinases should be particularly important for fungi with dramatic morphological changes, such as Ustilago maydis. This smut fungus switches from yeast to filamentous growth for plant infection, proliferates as a mycelium in planta, and forms teliospores for spreading. Here, we investigate the contribution of its four chitinolytic enzymes to the different morphological changes during the complete life cycle in a comprehensive study of deletion strains combined with biochemical and cell biological approaches. Interestingly, two chitinases act redundantly in cell separation during yeast growth. They mediate the degradation of remnant chitin in the fragmentation zone between mother and daughter cell. In contrast, even the complete lack of chitinolytic activity does not affect formation of the infectious filament, infection, biotrophic growth, or teliospore germination. Thus, unexpectedly we can exclude a major role for chitinolytic enzymes in morphogenesis or pathogenicity of U. maydis. Nevertheless, redundant activity of even two chitinases is essential for cell separation during saprophytic growth, possibly to improve nutrient access or spreading of yeast cells by wind or rain.

  12. Improved expression of single-chain antibodies in Ustilago maydis.

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    Sarkari, Parveen; Reindl, Michèle; Stock, Janpeter; Müller, Olaf; Kahmann, Regine; Feldbrügge, Michael; Schipper, Kerstin

    2014-12-10

    To produce the full repertoire of biopharmaceutical proteins, alternative expression platforms are required. Systems that enable secretion of the target protein are favored because this facilitates downstream processing. Ustilago maydis is a promising fungal model organism for future applications in protein expression. Recently, we described the exploitation of a novel unconventional secretion mechanism for the export of heterologous proteins. In this mode of secretion, the endochitinase Cts1 functions as a carrier for export with the main advantage of avoiding potentially harmful N-glycosylation. The major limitation until now was a low yield of secreted full-length protein. For optimization, we identified two bottlenecks: mRNA amount and extracellular proteolytic activity. By generating novel expression vectors harboring a strong constitutive promoter as well as eliminating harmful proteases, yields were increased significantly. A scFv antibody fragment against the cMyc epitope served as proof-of-principle and could be purified in its active, full-length form from the culture supernatant. Thus, we improved the novel expression system in U. maydis such that it can now be investigated with respect to other targets with potential applications for instance in diagnostics and medicine.

  13. Identification of O-mannosylated virulence factors in Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Alfonso Fernández-Álvarez

    Full Text Available The O-mannosyltransferase Pmt4 has emerged as crucial for fungal virulence in the animal pathogens Candida albicans or Cryptococcus neoformans as well as in the phytopathogenic fungus Ustilago maydis. Pmt4 O-mannosylates specific target proteins at the Endoplasmic Reticulum. Therefore a deficient O-mannosylation of these target proteins must be responsible for the loss of pathogenicity in pmt4 mutants. Taking advantage of the characteristics described for Pmt4 substrates in Saccharomyces cerevisiae, we performed a proteome-wide bioinformatic approach to identify putative Pmt4 targets in the corn smut fungus U. maydis and validated Pmt4-mediated glycosylation of candidate proteins by electrophoretic mobility shift assays. We found that the signalling mucin Msb2, which regulates appressorium differentiation upstream of the pathogenicity-related MAP kinase cascade, is O-mannosylated by Pmt4. The epistatic relationship of pmt4 and msb2 showed that both are likely to act in the same pathway. Furthermore, constitutive activation of the MAP kinase cascade restored appressorium development in pmt4 mutants, suggesting that during the initial phase of infection the failure to O-mannosylate Msb2 is responsible for the virulence defect of pmt4 mutants. On the other hand we demonstrate that during later stages of pathogenic development Pmt4 affects virulence independently of Msb2, probably by modifying secreted effector proteins. Pit1, a protein required for fungal spreading inside the infected leaf, was also identified as a Pmt4 target. Thus, O-mannosylation of different target proteins affects various stages of pathogenic development in U. maydis.

  14. Heterologous production and characterization of a chlorogenic acid esterase from Ustilago maydis with a potential use in baking.

    Science.gov (United States)

    Nieter, Annabel; Kelle, Sebastian; Takenberg, Meike; Linke, Diana; Bunzel, Mirko; Popper, Lutz; Berger, Ralf G

    2016-10-15

    Ustilago maydis, an edible mushroom growing on maize (Zea mays), is consumed as the food delicacy huitlacoche in Mexico. A chlorogenic acid esterase from this basidiomycete was expressed in good yields cultivating the heterologous host Pichia pastoris on the 5L bioreactor scale (reUmChlE; 45.9UL(-1)). In contrast to previously described chlorogenic acid esterases, the reUmChlE was also active towards feruloylated saccharides. The enzyme preferred substrates with the ferulic acid esterified to the O-5 position of arabinose residues, typical of graminaceous monocots, over the O-2 position of arabinose or the O-6 position of galactose residues. Determination of kcat/Km showed that the reUmChlE hydrolyzed chlorogenic acid 18-fold more efficiently than methyl ferulate, p-coumarate or caffeate. Phenolic acids were released by reUmChlE from natural substrates, such as destarched wheat bran, sugar beet pectin and coffee pulp. Treatment of wheat dough using reUmChlE resulted in a noticeable softening indicating a potential application of the enzyme in bakery and confectionery. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Degradation of the plant defence hormone salicylic acid by the biotrophic fungus Ustilago maydis.

    Science.gov (United States)

    Rabe, Franziska; Ajami-Rashidi, Ziba; Doehlemann, Gunther; Kahmann, Regine; Djamei, Armin

    2013-07-01

    Salicylic acid (SA) is a key plant defence hormone which plays an important role in local and systemic defence responses against biotrophic pathogens like the smut fungus Ustilago maydis. Here we identified Shy1, a cytoplasmic U. maydis salicylate hydroxylase which has orthologues in the closely related smuts Ustilago hordei and Sporisorium reilianum. shy1 is transcriptionally induced during the biotrophic stages of development but not required for virulence during seedling infection. Shy1 activity is needed for growth on plates with SA as a sole carbon source. The trigger for shy1 transcriptional induction is SA, suggesting the possibility of a SA sensing mechanism in this fungus.

  16. Comparative analysis of the Maize Smut Fungi Ustilago maydis and Sporisorium reilianum

    OpenAIRE

    Heinze, Bernadette

    2010-01-01

    The maize plant (Zea mays) is host to two closely related phytopathogenic fungi Ustilago maydis and Sporisorium reilianum. U. maydis infects all aerial parts of the plant, rapidly forming galls or tumours filled with spores. S. reilianum infects young seedlings, remains asymptomatic and grows systemically until it replaces the inflorescence with a mass of sooty spores. The identification of the factors responsible for the differen...

  17. Characterization of the largest effector gene cluster of Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Thomas Brefort

    2014-07-01

    Full Text Available In the genome of the biotrophic plant pathogen Ustilago maydis, many of the genes coding for secreted protein effectors modulating virulence are arranged in gene clusters. The vast majority of these genes encode novel proteins whose expression is coupled to plant colonization. The largest of these gene clusters, cluster 19A, encodes 24 secreted effectors. Deletion of the entire cluster results in severe attenuation of virulence. Here we present the functional analysis of this genomic region. We show that a 19A deletion mutant behaves like an endophyte, i.e. is still able to colonize plants and complete the infection cycle. However, tumors, the most conspicuous symptoms of maize smut disease, are only rarely formed and fungal biomass in infected tissue is significantly reduced. The generation and analysis of strains carrying sub-deletions identified several genes significantly contributing to tumor formation after seedling infection. Another of the effectors could be linked specifically to anthocyanin induction in the infected tissue. As the individual contributions of these genes to tumor formation were small, we studied the response of maize plants to the whole cluster mutant as well as to several individual mutants by array analysis. This revealed distinct plant responses, demonstrating that the respective effectors have discrete plant targets. We propose that the analysis of plant responses to effector mutant strains that lack a strong virulence phenotype may be a general way to visualize differences in effector function.

  18. LAMMER kinase contributes to genome stability in Ustilago maydis.

    Science.gov (United States)

    de Sena-Tomás, Carmen; Sutherland, Jeanette H; Milisavljevic, Mira; Nikolic, Dragana B; Pérez-Martín, José; Kojic, Milorad; Holloman, William K

    2015-09-01

    Here we report identification of the lkh1 gene encoding a LAMMER kinase homolog (Lkh1) from a screen for DNA repair-deficient mutants in Ustilago maydis. The mutant allele isolated results from a mutation at glutamine codon 488 to a stop codon that would be predicted to lead to truncation of the carboxy-terminal kinase domain of the protein. This mutant (lkh1(Q488*)) is highly sensitive to ultraviolet light, methyl methanesulfonate, and hydroxyurea. In contrast, a null mutant (lkh1Δ) deleted of the entire lkh1 gene has a less severe phenotype. No epistasis was observed when an lkh1(Q488*)rad51Δ double mutant was tested for genotoxin sensitivity. However, overexpressing the gene for Rad51, its regulator Brh2, or the Brh2 regulator Dss1 partially restored genotoxin resistance of the lkh1Δ and lkh1(Q488*) mutants. Deletion of lkh1 in a chk1Δ mutant enabled these double mutant cells to continue to cycle when challenged with hydroxyurea. lkh1Δ and lkh1(Q488*) mutants were able to complete the meiotic process but exhibited reduced heteroallelic recombination and aberrant chromosome segregation. The observations suggest that Lkh1 serves in some aspect of cell cycle regulation after DNA damage or replication stress and that it also contributes to proper chromosome segregation in meiosis.

  19. Characterization of the largest effector gene cluster of Ustilago maydis.

    Science.gov (United States)

    Brefort, Thomas; Tanaka, Shigeyuki; Neidig, Nina; Doehlemann, Gunther; Vincon, Volker; Kahmann, Regine

    2014-07-01

    In the genome of the biotrophic plant pathogen Ustilago maydis, many of the genes coding for secreted protein effectors modulating virulence are arranged in gene clusters. The vast majority of these genes encode novel proteins whose expression is coupled to plant colonization. The largest of these gene clusters, cluster 19A, encodes 24 secreted effectors. Deletion of the entire cluster results in severe attenuation of virulence. Here we present the functional analysis of this genomic region. We show that a 19A deletion mutant behaves like an endophyte, i.e. is still able to colonize plants and complete the infection cycle. However, tumors, the most conspicuous symptoms of maize smut disease, are only rarely formed and fungal biomass in infected tissue is significantly reduced. The generation and analysis of strains carrying sub-deletions identified several genes significantly contributing to tumor formation after seedling infection. Another of the effectors could be linked specifically to anthocyanin induction in the infected tissue. As the individual contributions of these genes to tumor formation were small, we studied the response of maize plants to the whole cluster mutant as well as to several individual mutants by array analysis. This revealed distinct plant responses, demonstrating that the respective effectors have discrete plant targets. We propose that the analysis of plant responses to effector mutant strains that lack a strong virulence phenotype may be a general way to visualize differences in effector function.

  20. Induction of infection in Teosinte (Zea diploperennis through the phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Martha Concepción Pérez Díaz

    2017-01-01

    Full Text Available Introduction: The corn and teosinte share morphological and molecular similarities latter being those that support the theory of teosinte (Zea parviglumis as its predecessor, both species are attacked by specific pathogens like Ustilago maydis. Objective: To analyze the infectious process that presents U. maydis on the variety of the teosinte Zea diploperennis. Materials and Methods: We used the strain of U. maydis FB-D12, which was kept a culture media rich in nutrients (CPES pH 7.0. Viable cells without morphological alterations to the inoculation method of puncture in teosinte seedlings were used. Monitoring of infection was carried out every 24 hours by measuring concentration of chlorophyll and plant tissue through microscopic observation Results: In the seedlings of Zea diploperennis inoculated with U. maydis the symptoms of the infection were presented, wilt and chlorosis in the leaves; The chlorosis was confirmed with the low concentration of chlorophyll 12 days later to the inoculation. In the microscopic observation of cuts of the tissue plant was found mycelium long and branched from the third day of the inoculation, until the appearance of tumors in seedlings of 45 days. Conclusions: The typical signs of infection with Ustilago maydis in the variety of teosinte Zea diploperennis do not differ from those reported for corn. Ustilago maydis presents its full life cycle within the plant, confirming that the diploperennis variety is susceptible.

  1. Isolation of UmRrm75, a gene involved in dimorphism and virulence of Ustilago maydis

    Science.gov (United States)

    Ustilago maydis displays dimorphic growth, alternating between a saprophytic haploid yeast form and a filamentous dikaryon, generated by mating of haploid cells and which is an obligate parasite. Induction of the dimorphic transition of haploid strains in vitro by change in ambient pH has been used...

  2. Brh2-Dss1 interplay enables properly controlled recombination in Ustilago maydis

    DEFF Research Database (Denmark)

    Kojic, Milorad; Zhou, Qingwen; Lisby, Michael;

    2005-01-01

    Brh2, the BRCA2 homolog in Ustilago maydis, functions in recombinational repair of DNA damage by regulating Rad51 and is, in turn, regulated by Dss1. Dss1 is not required for Brh2 stability in vivo, nor for Brh2 to associate with Rad51, but is required for formation of green fluorescent protein...

  3. The metabolome and transcriptome of the interaction between Ustilago maydis and Fusarium verticillioides in vitro

    Science.gov (United States)

    The metabolome and transcriptome of the maize-infecting fungi Ustilago maydis and Fusarium verticillioides were analyzed as the two fungi interact. Both fungi were grown for seven days in liquid medium alone or together in order to study how this interaction changes their metabolomic and transcripto...

  4. Aflatoxin and Fumonisin in corn (Zea mays) infected by common smut Ustilago maydis

    Science.gov (United States)

    Corn infected with Ustilago maydis (common smut) produces galls that are valued food in certain cultures, but may be contaminated with mycotoxins. Field studies conducted in Elizabeth, Mississippi used near isogenic Bt and non-Bt corn hybrids. The levels of aflatoxin and fumonisin were determined i...

  5. Rec2 Interplay with both Brh2 and Rad51 Balances Recombinational Repair in Ustilago maydis

    DEFF Research Database (Denmark)

    Kojic, M.; Zhou, Q.; Lisby, M.;

    2006-01-01

    Rec2 is the single Rad51 paralog in Ustilago maydis. Here, we find that Rec2 is required for radiation-induced Rad51 nuclear focus formation but that Rec2 foci form independently of Rad51 and Brh2. Brh2 foci also form in the absence of Rad51 and Rec2. By coprecipitation from cleared extracts...

  6. Virulence of the maize smut Ustilago maydis is shaped by organ-specific effectors.

    Science.gov (United States)

    Schilling, Lena; Matei, Alexandra; Redkar, Amey; Walbot, Virginia; Doehlemann, Gunther

    2014-10-01

    With the exception of Ustilago maydis, smut fungi infecting monocotyledonous hosts systemically colonize infected plants and cause symptoms exclusively in the inflorescences. Ustilago may disinfects primordia of all aerial organs of maize (Zea mays L.) and results in the formation of large plant tumours. Previously, we have found that U. maydis infection of seedling leaves, adult leaves and tassels causes organ-specific transcriptional changes in both the pathogen and the host. Of particular interest, U. may disgenes encoding secreted proteins are differentially expressed depending on the colonized maize organ. Therefore, we hypothesized that the fungus secretes virulence-related proteins (effectors)that act in an organ-specific manner. Here, we present the identification and functional characterization of 20 presumptive organ-specific U. maydis effector genes. Ustilago maydis deletion strains for these genes were generated and tested for infectivity of maize seedling leaves and tassels. This approach identified 11 effector genes required for the full virulence of U. maydis. In nine cases, virulence was only affected in one of the tested plant organs. These results demonstrate that individual fungal effector proteins contribute to fungal virulence in an organ-specific manner.

  7. Identification of Ustilago maydis Aurora kinase as a novel antifungal target.

    Science.gov (United States)

    Tückmantel, Sandra; Greul, Jörg N; Janning, Petra; Brockmeyer, Andreas; Grütter, Christian; Simard, Jeffrey R; Gutbrod, Oliver; Beck, Michael E; Tietjen, Klaus; Rauh, Daniel; Schreier, Peter H

    2011-09-16

    Infestation of crops by pathogenic fungi has continued to have a major impact by reducing yield and quality, emphasizing the need to identify new targets and develop new agents to improve methods of crop protection. Here we present Aurora kinase from the phytopathogenic fungus Ustilago maydis as a novel target for N-substituted diaminopyrimidines, a class of small-molecule kinase inhibitors. We show that Aurora kinase is essential in U. maydis and that diaminopyrimidines inhibit its activity in vitro. Furthermore, we observed an overall good correlation between in vitro inhibition of Aurora kinase and growth inhibition of diverse fungi in vivo. In vitro inhibition assays with Ustilago and human Aurora kinases indicate that some compounds of the N-substituted diaminopyrimidine class show specificity for the Ustilago enzyme, thus revealing their potential as selective fungicides.

  8. Conservation of the Ustilago maydis effector See1 in related smuts.

    Science.gov (United States)

    Redkar, Amey; Villajuana-Bonequi, Mitzi; Doehlemann, Gunther

    2015-01-01

    Ustilago maydis is a biotrophic fungus that induces formation of tumors in maize (Zea mays L). In a recent study we identified See1 (Seedling efficient effector 1) as an U. maydis organ-specific effector required for tumor formation in leaves. See1 is required for U. maydis induced reactivation of plant DNA synthesis during leaf tumor progression. The protein is secreted from biotrophic hyphae and localizes to the cytoplasm and nucleus of plant cell. See1 interacts with maize SGT1, a cell cycle and immune regulator, interfering with its MAPK-triggered phosphorylation. Here, we present new data on the conservation of See1 in other closely related smuts and experimental data on the functionality of See1 ortholog in Ustilago hordei, the causal agent of barley covered smut disease.

  9. Genome editing in Ustilago maydis using the CRISPR-Cas system.

    Science.gov (United States)

    Schuster, Mariana; Schweizer, Gabriel; Reissmann, Stefanie; Kahmann, Regine

    2016-04-01

    This communication describes the establishment of the type II bacterial CRISPR-Cas9 system to efficiently disrupt target genes in the fungal maize pathogen Ustilago maydis. A single step transformation of a self-replicating plasmid constitutively expressing the U. maydis codon-optimized cas9 gene and a suitable sgRNA under control of the U. maydis U6 snRNA promoter was sufficient to induce genome editing. On average 70% of the progeny of a single transformant were disrupted within the respective b gene. Without selection the self-replicating plasmid was lost rapidly allowing transient expression of the CRISPR-Cas9 system to minimize potential long-term negative effects of Cas9. This technology will be an important advance for the simultaneous disruption of functionally redundant genes and gene families to investigate their contribution to virulence of U. maydis.

  10. Ustilago maydis: ascenso de un hongo mexicano de la gastronomía local al mundo científico

    Directory of Open Access Journals (Sweden)

    José Ruiz Herrera

    2008-01-01

    Full Text Available En el presente trabajo se describe la naturaleza del hongo Ustilago maydis, su papel como causante de una enfermedad del maíz, su uso en la cocina mexicana y las condiciones que lo llevaron a convertirse en material de estudio científico, además de algunas características descollantes de su fisiología, genética y bioquímica. U. maydis es un hongo patógeno biotrófico específico del maíz, y el teozintle, siendo el agente causal del "huitlacoche" o carbón común, una enfermedad de distribución mundial que bajo ciertas condiciones puede causar severos daños económicos. Sin embargo, en México, las mazorcas infectadas han sido un alimento usado en la cocina tradicional desde la época pre-colombina, y actualmente también en la alta gastronomía del país y el extranjero. El hongo y sus huéspedes son nativos de la parte central de México, y fueron introducidos en Europa por los españoles, donde la enfermedad fue considerada como una alteración fisiológica de la planta. Su naturaleza como infección fúngica fue reconocida solo hasta la segunda mitad del siglo XIX. U. maydis se introdujo a los laboratorios de investigación a principios del Siglo XX, convirtiéndose en un modelo clásico para el estudio de las bases de la patogénesis fúngica en plantas, de la especificidad en el apareamiento y de la recombinación genética, entre otras. Esto fue debido a su facilidad de manejo, su corto ciclo de vida, y el ser posible su análisis mediante métodos de genética clásica y molecular. El ciclo de vida del hongo es complicado alternando formas de levadura haploide saprofítica y micelio dicariótico infeccioso. Este ciclo sexual solo se completa en su huésped natural, estando bajo el control de dos loci de apareamiento que controlan la fusión celular y la patogénesis. En condiciones de cultivo axénico, hemos demostrado que el hongo es capaz de infectar plantas filogenéticamente distantes del maíz; y hemos desarrollado

  11. Kinesin-3 in the basidiomycete Ustilago maydis transports organelles along the entire microtubule array.

    Science.gov (United States)

    Steinberg, Gero

    2015-01-01

    The molecular motor kinesin-3 transports early endosomes along microtubules in filamentous fungi. It was reported that kinesin-3 from the ascomycete fungi Aspergillus nidulans and Neurospora crassa use a subset of post-translationally modified and more stable microtubules. Here, I show that kinesin-3 from the basidiomycete Ustilago maydis moves along all hyphal microtubules. This difference is likely due to variation in cell cycle control and associated organization of the microtubule array.

  12. Assessment of Ustilago maydis as a fungal model for root infection studies.

    Science.gov (United States)

    Mazaheri-Naeini, Mahta; Sabbagh, Seyed Kazem; Martinez, Yves; Séjalon-Delmas, Nathalie; Roux, Christophe

    2015-03-01

    Ustilago maydis is a fungus infecting aerial parts of maize to form smutted galls. Due to its interest as a genetic tool in plant pathology, we evaluated its ability to penetrate into plant roots. The fungus can penetrate between epidermic root cells, forming inter and intracellular pseudohyphae. Root infection didn't provoke gall formation on the maize lines tested, and targeted PCR detection showed that U. maydis, unlike the other maize smut fungus Sporisorium reilianum, has a weak aptitude to grow from the roots up to the aerial part of maize. We also observed that U. maydis can infect Medicago truncatula hairy roots as an alternative host. This plant species is a model host to study root symbiosis, and this pathosystem can provide new insights on root-microbe interactions. Considering that U. maydis could be a soil fungus, we tested its responsiveness to GR24, a strigolactone analogue. Strigolactones are root exuded molecules which activate mitochondrial metabolism of arbuscular mycorrhizal (AM) fungi. Physiologic and molecular analysis revealed that GR24 also increases cell respiration of U. maydis. This result points out that strigolactones could have an incidence on several rhizospheric microbes. These data provide evidences that the biotrophic pathogen U. maydis has to be considered for studying root infection.

  13. Influence of chitosan and its derivatives on cell development and physiology of Ustilago maydis.

    Science.gov (United States)

    Olicón-Hernández, Dario Rafael; Hernández-Lauzardo, Ana N; Pardo, Juan Pablo; Peña, Antonio; Velázquez-del Valle, Miguel G; Guerra-Sánchez, Guadalupe

    2015-08-01

    Ustilago maydis, a dimorphic fungus causing corn smut disease, serves as an excellent model to study different aspects of cell development. This study shows the influence of chitosan, oligochitosan and glycol chitosan on cell growth and physiology of U. maydis. These biological macromolecules affected the cell growth of U. maydis. In particular, it was found that chitosan completely inhibited U. maydis growth at 1mg/mL concentration. Microscopic studies revealed swellings on the surface of the cells treated with the polymers, and chitosan caused complete destruction of the membrane and formation of vesicles on the periphery of the cell. Oligochitosan and chitosan caused changes in oxygen consumption, K(+) efflux and H(+)-ATPase activity. Oligochitosan induced a faster consumption of oxygen in the cells, while glycol chitosan provoked slower oxygen consumption. It is noteworthy that chitosan completely inhibited the fungal respiratory activity. The strongest effects were exhibited by chitosan in all evaluated aspects. These findings showed high sensitivity of U. maydis to chitosan and provided evidence for antifungal effects of chitosan derivatives. To our knowledge, this is a first report showing that chitosan and its derivatives affect the cell morphology and physiological processes in U. maydis.

  14. Metabolome and transcriptome of the interaction between Ustilago maydis and Fusarium verticillioides in vitro.

    Science.gov (United States)

    Jonkers, Wilfried; Rodriguez Estrada, Alma E; Lee, Keunsub; Breakspear, Andrew; May, Georgiana; Kistler, H Corby

    2012-05-01

    The metabolome and transcriptome of the maize-infecting fungi Ustilago maydis and Fusarium verticillioides were analyzed as the two fungi interact. Both fungi were grown for 7 days in liquid medium alone or together in order to study how this interaction changes their metabolomic and transcriptomic profiles. When grown together, decreased biomass accumulation occurs for both fungi after an initial acceleration of growth compared to the biomass changes that occur when grown alone. The biomass of U. maydis declined most severely over time and may be attributed to the action of F. verticillioides, which secretes toxic secondary metabolites and expresses genes encoding adhesive and cell wall-degrading proteins at higher levels than when grown alone. U. maydis responds to cocultivation by expressing siderophore biosynthetic genes and more highly expresses genes potentially involved in toxin biosynthesis. Also, higher expression was noted for clustered genes encoding secreted proteins that are unique to U. maydis and that may play a role during colonization of maize. Conversely, decreased gene expression was seen for U. maydis genes encoding the synthesis of ustilagic acid, mannosylerythritol D, and another uncharacterized metabolite. Ultimately, U. maydis is unable to react efficiently to the toxic response of F. verticillioides and proportionally loses more biomass. This in vitro study clarifies potential mechanisms of antagonism between these two fungi that also may occur in the soil or in maize, niches for both fungi where they likely interact in nature.

  15. Metamorphosis of the Basidiomycota Ustilago maydis: transformation of yeast-like cells into basidiocarps.

    Science.gov (United States)

    Cabrera-Ponce, José L; León-Ramírez, Claudia G; Verver-Vargas, Aurora; Palma-Tirado, Lourdes; Ruiz-Herrera, José

    2012-10-01

    Ustilago maydis (DC) Cda., a phytopathogenic Basidiomycota, is the causal agent of corn smut. During its life cycle U. maydis alternates between a yeast-like, haploid nonpathogenic stage, and a filamentous, dikaryotic pathogenic form that invades the plant and induces tumor formation. As all the members of the Subphylum Ustilaginomycotina, U. maydis is unable to form basidiocarps, instead it produces teliospores within the tumors that germinate forming a septate basidium (phragmobasidium). We have now established conditions allowing a completely different developmental program of U. maydis when grown on solid medium containing auxins in dual cultures with maize embryogenic calli. Under these conditions U. maydis forms large hemi-spheroidal structures with all the morphological and structural characteristics of gastroid-type basidiocarps. These basidiocarps are made of three distinct hyphal layers, the most internal of which (hymenium) contains non-septate basidia (holobasidia) from which four basidiospores develop. In basidiocarps meiosis and genetic recombination occur, and meiotic products (basidiospores) segregate in a Mendelian fashion. These results are evidence of sexual cycle completion of an Ustilaginomycotina in vitro, and the demonstration that, besides its quasi-obligate biotrophic pathogenic mode of life, U. maydis possesses the genetic program to form basidiocarps as occurs in saprophytic Basidiomycota species.

  16. Inhibition of polyamine oxidase activity affects tumor development during the maize-Ustilago maydis interaction.

    Science.gov (United States)

    Jasso-Robles, Francisco Ignacio; Jiménez-Bremont, Juan Francisco; Becerra-Flora, Alicia; Juárez-Montiel, Margarita; Gonzalez, María Elisa; Pieckenstain, Fernando Luis; García de la Cruz, Ramón Fernando; Rodríguez-Kessler, Margarita

    2016-05-01

    Ustilago maydis is a biotrophic plant pathogenic fungus that leads to tumor development in the aerial tissues of its host, Zea mays. These tumors are the result of cell hypertrophy and hyperplasia, and are accompanied by the reprograming of primary and secondary metabolism of infected plants. Up to now, little is known regarding key plant actors and their role in tumor development during the interaction with U. maydis. Polyamines are small aliphatic amines that regulate plant growth, development and stress responses. In a previous study, we found substantial increases of polyamine levels in tumors. In the present work, we describe the maize polyamine oxidase (PAO) gene family, its contribution to hydrogen peroxide (H2O2) production and its possible role in tumor development induced by U. maydis. Histochemical analysis revealed that chlorotic lesions and maize tumors induced by U. maydis accumulate H2O2 to significant levels. Maize plants inoculated with U. maydis and treated with the PAO inhibitor 1,8-diaminooctane exhibit a notable reduction of H2O2 accumulation in infected tissues and a significant drop in PAO activity. This treatment also reduced disease symptoms in infected plants. Finally, among six maize PAO genes only the ZmPAO1, which encodes an extracellular enzyme, is up-regulated in tumors. Our data suggest that H2O2 produced through PA catabolism by ZmPAO1 plays an important role in tumor development during the maize-U. maydis interaction.

  17. Experimental approaches to investigate effector translocation into host cells in the Ustilago maydis/maize pathosystem.

    Science.gov (United States)

    Tanaka, Shigeyuki; Djamei, Armin; Presti, Libera Lo; Schipper, Kerstin; Winterberg, Sarah; Amati, Simone; Becker, Dirk; Büchner, Heike; Kumlehn, Jochen; Reissmann, Stefanie; Kahmann, Regine

    2015-01-01

    The fungus Ustilago maydis is a pathogen that establishes a biotrophic interaction with Zea mays. The interaction with the plant host is largely governed by more than 300 novel, secreted protein effectors, of which only four have been functionally characterized. Prerequisite to examine effector function is to know where effectors reside after secretion. Effectors can remain in the extracellular space, i.e. the plant apoplast (apoplastic effectors), or can cross the plant plasma membrane and exert their function inside the host cell (cytoplasmic effectors). The U. maydis effectors lack conserved motifs in their primary sequences that could allow a classification of the effectome into apoplastic/cytoplasmic effectors. This represents a significant obstacle in functional effector characterization. Here we describe our attempts to establish a system for effector classification into apoplastic and cytoplasmic members, using U. maydis for effector delivery.

  18. The promoter of the glucoamylase-encoding gene of Aspergillus niger functions in Ustilago maydis

    Energy Technology Data Exchange (ETDEWEB)

    Smith, T.L. (Dept. of Agriculture, Madison, WI (United States) Univ. of Wisconsin, Madison (United States)); Gaskell, J.; Cullen, D. (Dept. of Agriculture, Madison, WI (United States)); Berka, R.M.; Yang, M.; Henner, D.J. (Genentech Inc., San Francisco, CA (United States))

    1990-01-01

    Promoter sequences from the Aspergillus niger glucoamylase-encoding gene (glaA) were linked to the bacterial hygromycin (Hy) phosphotransferase-encoding gene (hph) and this chimeric marker was used to select Hy-resistant (Hy[sup R]) Ustilago maydis transformants. This is an example of an Ascomycete promoter functioning in a Basidiomycete. Hy[sup R] transformants varied with respect to copy number of integrated vector, mitotic stability, and tolerance to Hy. Only 216 bp of glaA promoter sequence is required for expression in U. maydis but this promoter is not induced by starch as it is in Aspergillus spp. The transcription start points are the same in U. maydis and A. niger.

  19. The Corn Smut ('Huitlacoche') as a New Platform for Oral Vaccines.

    Science.gov (United States)

    Juárez-Montiel, Margarita; Romero-Maldonado, Andrea; Monreal-Escalante, Elizabeth; Becerra-Flora, Alicia; Korban, Schuyler S; Rosales-Mendoza, Sergio; Jiménez-Bremont, Juan Francisco

    2015-01-01

    The development of new alternative platforms for subunit vaccine production is a priority in the biomedical field. In this study, Ustilago maydis, the causal agent of common corn smut or 'huitlacoche'has been genetically engineered to assess expression and immunogenicity of the B subunit of the cholera toxin (CTB), a relevant immunomodulatory agent in vaccinology. An oligomeric CTB recombinant protein was expressed in corn smut galls at levels of up to 1.3 mg g-1 dry weight (0.8% of the total soluble protein). Mice orally immunized with 'huitlacoche'-derived CTB showed significant humoral responses that were well-correlated with protection against challenge with the cholera toxin (CT). These findings demonstrate the feasibility of using edible corn smut as a safe, effective, and low-cost platform for production and delivery of a subunit oral vaccine. The implications of this platform in the area of molecular pharming are discussed.

  20. The Corn Smut ('Huitlacoche' as a New Platform for Oral Vaccines.

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    Margarita Juárez-Montiel

    Full Text Available The development of new alternative platforms for subunit vaccine production is a priority in the biomedical field. In this study, Ustilago maydis, the causal agent of common corn smut or 'huitlacoche'has been genetically engineered to assess expression and immunogenicity of the B subunit of the cholera toxin (CTB, a relevant immunomodulatory agent in vaccinology. An oligomeric CTB recombinant protein was expressed in corn smut galls at levels of up to 1.3 mg g-1 dry weight (0.8% of the total soluble protein. Mice orally immunized with 'huitlacoche'-derived CTB showed significant humoral responses that were well-correlated with protection against challenge with the cholera toxin (CT. These findings demonstrate the feasibility of using edible corn smut as a safe, effective, and low-cost platform for production and delivery of a subunit oral vaccine. The implications of this platform in the area of molecular pharming are discussed.

  1. Transcriptomic analysis of Ustilago maydis infecting Arabidopsis reveals important aspects of the fungus pathogenic mechanisms.

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    Martínez-Soto, Domingo; Robledo-Briones, Angélica M; Estrada-Luna, Andrés A; Ruiz-Herrera, José

    2013-08-01

    Transcriptomic and biochemical analyses of the experimental pathosystem constituted by Ustilago maydis and Arabidopsis thaliana were performed. Haploid or diploid strains of U. maydis inoculated in A. thaliana plantlets grew on the surface and within the plant tissues in the form of mycelium, inducing chlorosis, anthocyanin formation, malformations, necrosis and adventitious roots development, but not teliospores. Symptoms were more severe in plants inoculated with the haploid strain which grew more vigorously than the diploid strain. RNA extracted at different times post-infection was used for hybridization of one-channel microarrays that were analyzed focusing on the fungal genes involved in the general pathogenic process, biogenesis of the fungal cell wall and the secretome. In total, 3,537 and 3,299 genes were differentially expressed in the haploid and diploid strains, respectively. Differentially expressed genes were related to different functional categories and many of them showed a similar regulation occurring in U. maydis infecting maize. Our data suggest that the haploid strain behaves as a necrotrophic pathogen, whereas the diploid behaves as a biotrophic pathogen. The results obtained are evidence of the usefulness of the U. maydis-A. thaliana pathosystem for the analysis of the pathogenic mechanisms of U. maydis.

  2. Analysis of a polygalacturonase gene of Ustilago maydis and characterization of the encoded enzyme.

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    Castruita-Domínguez, José P; González-Hernández, Sandra E; Polaina, Julio; Flores-Villavicencio, Lérida L; Alvarez-Vargas, Aurelio; Flores-Martínez, Alberto; Ponce-Noyola, Patricia; Leal-Morales, Carlos A

    2014-05-01

    Ustilago maydis is a pathogenic fungus that produces the corn smut. It is a biotrophic parasite that depends on living plant tissues for its proliferation and development. Polygalacturonases are secreted by pathogens to solubilize the plant cell-wall and are required for pathogen virulence. In this paper, we report the isolation of a U. maydis polygalacturonase gene (Pgu1) and the functional and structural characterization of the encoded enzyme. The U. maydis Pgu1 gene is expressed when the fungus is grown in liquid culture media containing different carbon sources. In plant tissue, the expression increased as a function of incubation time. Pgu1 gene expression was detected during plant infection around 10 days post-infection with U. maydis FB-D12 strain in combination with teliospore formation. Synthesis and secretion of active recombinant PGU1 were achieved using Pichia pastoris, the purified enzyme had a optimum temperature of 34 °C, optimum pH of 4.5, a Km of 57.84 g/L for polygalacturonic acid, and a Vmax of 28.9 µg/min mg. Structural models of PGU1 based on homologous enzymes yielded a typical right-handed β-helix fold of pectinolytic enzymes classified in the glycosyl hydrolases family 28, and the U. maydis PGU1 is related with endo rather than exo polygalacturonases.

  3. Cleavage of resveratrol in fungi: characterization of the enzyme Rco1 from Ustilago maydis.

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    Brefort, Thomas; Scherzinger, Daniel; Limón, M Carmen; Estrada, Alejandro F; Trautmann, Danika; Mengel, Carina; Avalos, Javier; Al-Babili, Salim

    2011-02-01

    Ustilago maydis, the causative agent of corn smut disease, contains two genes encoding members of the carotenoid cleavage oxygenase family, a group of enzymes that cleave double bonds in different substrates. One of them, Cco1, was formerly identified as a β-carotene cleaving enzyme. Here we elucidate the function of the protein encoded by the second gene, termed here as Ustilago maydis Resveratrol cleavage oxygenase 1 (Um Rco1). In vitro incubations of heterologously expressed and purified UM Rco1 with different carotenoid and stilbene substrates demonstrate that it cleaves the interphenyl Cα-Cβ double bond of the phytoalexin resveratrol and its derivative piceatannol. Um Rco1 exhibits a high degree of substrate specificity, as suggested by the lack of activity on carotenoids and the other resveratrol-related compounds tested. The activity of Um Rco1 was confirmed by incubation of U. maydis rco1 deletion and over-expression strains with resveratrol. Furthermore, treatment with resveratrol resulted in striking alterations of cell morphology. However, pathogenicity assays indicated that Um rco1 is largely dispensable for biotrophic development. Our work reveals Um Rco1 as the first eukaryotic resveratrol cleavage enzyme identified so far. Moreover, Um Rco1 represents a subfamily of fungal enzymes likely involved in the degradation of stilbene compounds, as suggested by the cleavage of resveratrol by homologs from Aspergillus fumigatus, Chaetomium globosum and Botryotinia fuckeliana.

  4. An unusual MAP kinase is required for efficient penetration of the plant surface by Ustilago maydis

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    Brachmann, Andreas; Schirawski, Jan; Müller, Philip; Kahmann, Regine

    2003-01-01

    In Ustilago maydis, pathogenic development is controlled by a heterodimer of the two homeodomain proteins bW and bE. We have identified by RNA fingerprinting a b-regulated gene, kpp6, which encodes an unusual MAP kinase. Kpp6 is similar to a number of other fungal MAP kinases involved in mating and pathogenicity, but contains an additional N-terminal domain unrelated to other proteins. Transcription of the kpp6 gene yields two transcripts differing in length, but encoding proteins of identica...

  5. Septins from the phytopathogenic fungus Ustilago maydis are required for proper morphogenesis but dispensable for virulence.

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    Isabel Alvarez-Tabarés

    Full Text Available BACKGROUND: Septins are a highly conserved family of GTP-binding proteins involved in multiple cellular functions, including cell division and morphogenesis. Studies of septins in fungal cells underpin a clear correlation between septin-based structures and fungal morphology, providing clues to understand the molecular frame behind the varied morphologies found in fungal world. METHODOLOGY/PRINCIPAL FINDINGS: Ustilago maydis genome has the ability to encode four septins. Here, using loss-of-function as well as GFP-tagged alleles of these septin genes, we investigated the roles of septins in the morphogenesis of this basidiomycete fungus. We described that septins in U. maydis could assemble into at least three different structures coexisting in the same cell: bud neck collars, band-like structures at the growing tip, and long septin fibers that run from pole to pole near the cell cortex. We also found that in the absence of septins, U. maydis cells lost their elongated shape, became wider at the central region and ended up losing their polarity, pointing to an important role of septins in the morphogenesis of this fungus. These morphological defects were alleviated in the presence of an osmotic stabilizer suggesting that absence of septins affected the proper formation of the cell wall, which was coherent with a higher sensitivity of septin defective cells to drugs that affect cell wall construction as well as exocytosis. As U. maydis is a phytopathogen, we analyzed the role of septins in virulence and found that in spite of the described morphological defects, septin mutants were virulent in corn plants. CONCLUSIONS/SIGNIFICANCE: Our results indicated a major role of septins in morphogenesis in U. maydis. However, in contrast to studies in other fungal pathogens, in which septins were reported to be necessary during the infection process, we found a minor role of septins during corn infection by U. maydis.

  6. Ustilago maydis accumulates beta-carotene at levels determined by a retinal-forming carotenoid oxygenase.

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    Estrada, Alejandro F; Brefort, Thomas; Mengel, Carina; Díaz-Sánchez, Violeta; Alder, Adrian; Al-Babili, Salim; Avalos, Javier

    2009-10-01

    The basidiomycete Ustilago maydis, the causative agent of corn smut disease, has emerged as a model organism for dimorphism and fungal phytopathogenicity. In this work, we line out the key conserved enzymes for beta-carotene biosynthesis encoded by the U. maydis genome and show that this biotrophic fungus accumulates beta-carotene. The amount of this pigment depended on culture pH and aeration but was not affected by light and was not increased by oxidative stress. Moreover, we identified the U. maydis gene, cco1, encoding a putative beta-carotene cleavage oxygenase. Heterologous overexpression and in vitro analyses of purified enzyme demonstrated that Cco1 catalyzes the symmetrical cleavage of beta-carotene to yield two molecules of retinal. Analyses of beta-carotene and retinal contents in U. maydiscco1 deletion and over-expression strains confirmed the enzymatic function of Cco1, and revealed that Cco1 determines the beta-carotene content. Our data indicate that carotenoid biosynthesis in U. maydis is carried out to provide retinal rather than to deliver protective pigments. The U. maydis genome also encodes three potential opsins, a family of photoactive proteins that use retinal as chromophore. Two opsin genes showed different light-regulated expression patterns, suggesting specialized roles in photobiology, while no mRNA was detected for the third opsin gene in the same experiments. However, deletion of the cco1 gene, which should abolish function of all the retinal-dependent opsins, did not affect growth, morphology or pathogenicity, suggesting that retinal and opsin proteins play no relevant role in U. maydis under the tested conditions.

  7. Cloning of the PYR3 gene of Ustilago maydis and its use in DNA transformation

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    Banks, G.R.; Taylor, S.Y. (National Institute for Medical Research, London (England))

    1988-12-01

    The Ustilago maydis PYR3 gene encoding dihydroorotase activity was cloned by direct complementation of Escherichia coli pyrC mutations. PYR3 transformants of E. coli pyrC mutants expressed homologous transcripts of a variety of sizes and regained dihydroorotase activity. PYR3 also complemented Saccharomyces cerevisiae ura4 mutations, and again multiple transcripts were expressed in transformants, and enzyme activity was regained. A 1.25-kilobase poly(rA)+ PYR3 transcript was detected in U. maydis itself. Linear DNA carrying the PYR3 gene transformed a U. maydis pyr3-1 pyrimidine auxotroph to prototrophy. Hybridization analysis revealed that three different types of transformants could be generated, depending on the structure of the transforming DNA used. The first type involved exchange of chromosomal mutant gene sequences with the cloned wild-type plasmid sequences. A second type had integrated linear transforming DNA at the chromosomal PYR3 locus, probably via a single crossover event. The third type had integrated transforming DNA sequences at multiple sites in the U. maydis genome. In the last two types, tandemly reiterated copies of the transforming DNA were found to have been integrated. All three types had lost the sensitivity of the parental pyr3-1 mutant to UV irradiation. They had also regained dihydroorotase activity, although its level did not correlate with the PYR3 gene copy number.

  8. Sho1 and Msb2-related proteins regulate appressorium development in the smut fungus Ustilago maydis.

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    Lanver, Daniel; Mendoza-Mendoza, Artemio; Brachmann, Andreas; Kahmann, Regine

    2010-06-01

    The dimorphic fungus Ustilago maydis switches from budding to hyphal growth on the plant surface. In response to hydrophobicity and hydroxy fatty acids, U. maydis develops infection structures called appressoria. Here, we report that, unlike in Saccharomyces cerevisiae and other fungi where Sho1 (synthetic high osmolarity sensitive) and Msb2 (multicopy suppressor of a budding defect) regulate stress responses and pseudohyphal growth, Sho1 and Msb2-like proteins play a key role during appressorium differentiation in U. maydis. Sho1 was identified through a two-hybrid screen as an interaction partner of the mitogen-activated protein (MAP) kinase Kpp6. Epistasis analysis revealed that sho1 and msb2 act upstream of the MAP kinases kpp2 and kpp6. Furthermore, Sho1 was shown to destabilize Kpp6 through direct interaction with the unique N-terminal domain in Kpp6, indicating a role of Sho1 in fine-tuning Kpp6 activity. Morphological differentiation in response to a hydrophobic surface was strongly attenuated in sho1 msb2 mutants, while hydroxy fatty acid-induced differentiation was unaffected. These data suggest that Sho1 and the transmembrane mucin Msb2 are involved in plant surface sensing in U. maydis.

  9. Response to osmotic stress and temperature of the fungus Ustilago maydis.

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    Salmerón-Santiago, Karina Gabriela; Pardo, Juan Pablo; Flores-Herrera, Oscar; Mendoza-Hernández, Guillermo; Miranda-Arango, Manuel; Guerra-Sánchez, Guadalupe

    2011-10-01

    Ustilago maydis is a fungal pathogen which is exposed during its life cycle to both abiotic and biotic stresses before and after the infection of maize. To cope with extreme environmental changes, microorganisms usually accumulate the disaccharide trehalose. We have investigated both the accumulation of trehalose and the activity of trehalase during the adaptation of U. maydis haploid cells to thermal, sorbitol, and NaCl stresses. Sorbitol and sodium chloride induced sustained accumulation of trehalose, while a transient increase was observed under heat stress. Sorbitol stressed cells showed higher trehalase activity compared with control cells and to those stressed by NaCl and high temperature. Addition of cycloheximide, a protein synthesis inhibitor, did not affect the trehalose accumulation during the first 15 min, but basal levels of trehalose were reached after the second period of 15 min. The proteomic analysis of the response of U. maydis to temperature, sorbitol, and salt stresses indicated a complex pattern which highlights the change of 18 proteins involved in carbohydrate and amino acid metabolism, protein folding, redox regulation, ion homeostasis, and stress response. We hypothesize that trehalose accumulation during sorbitol stress in U. maydis might be related to the adaptation of this organism during plant infection.

  10. Life without putrescine: disruption of the gene-encoding polyamine oxidase in Ustilago maydis odc mutants.

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    Valdés-Santiago, Laura; Guzmán-de-Peña, Doralinda; Ruiz-Herrera, José

    2010-11-01

    In previous communications the essential role of spermidine in Ustilago maydis was demonstrated by means of the disruption of the genes encoding ornithine decarboxylase (ODC) and spermidine synthase (SPE). However, the assignation of specific roles to each polyamine in different cellular functions was not possible because the spermidine added to satisfy the auxotrophic requirement of odc/spe double mutants is partly back converted into putrescine. In this study, we have approached this problem through the disruption of the gene-encoding polyamine oxidase (PAO), required for the conversion of spermidine into putrescine, and the construction of odc/pao double mutants that were unable to synthesize putrescine by either ornithine decarboxylation or retroconversion from spermidine. Phenotypic analysis of the mutants provided evidence that putrescine is only an intermediary in spermidine biosynthesis, and has no direct role in cell growth, dimorphic transition, or any other vital function of U. maydis. Nevertheless, our results show that putrescine may play a role in the protection of U. maydis against salt and osmotic stress, and possibly virulence. Evidence was also obtained that the retroconversion of spermidine into putrescine is not essential for U. maydis growth but may be important for its survival under natural conditions.

  11. Elucidation of the complete ferrichrome A biosynthetic pathway in Ustilago maydis.

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    Winterberg, Britta; Uhlmann, Stefanie; Linne, Uwe; Lessing, Franziska; Marahiel, Mohamed A; Eichhorn, Heiko; Kahmann, Regine; Schirawski, Jan

    2010-03-01

    Iron is an important element for many essential processes in living organisms. To acquire iron, the basidiomycete Ustilago maydis synthesizes the iron-chelating siderophores ferrichrome and ferrichrome A. The chemical structures of these siderophores have been elucidated long time ago but so far only two enzymes involved in their biosynthesis have been described. Sid1, an ornithine monoxygenase, is needed for the biosynthesis of both siderophores, and Sid2, a non-ribosomal peptide synthetase (NRPS), is involved in ferrichrome generation. In this work we identified four novel enzymes, Fer3, Fer4, Fer5 and Hcs1, involved in ferrichrome A biosynthesis in U. maydis. By HPLC-MS analysis of siderophore accumulation in culture supernatants of deletion strains, we show that Fer3, an NRPS, Fer4, an enoyl-coenzyme A (CoA)-hydratase, and Fer5, an acylase, are required for ferrichrome A production. We demonstrate by conditional expression of the hydroxymethyl glutaryl (HMG)-CoA synthase Hcs1 in U. maydis that HMG-CoA is an essential precursor for ferrichrome A. In addition, we heterologously expressed and purified Hcs1, Fer4 and Fer5, and demonstrated the enzymatic activities by in vitro experiments. Thus, we describe the first complete fungal siderophore biosynthetic pathway by functionally characterizing four novel genes responsible for ferrichrome A biosynthesis in U. maydis.

  12. Metallomics approach to trace element analysis in ustilago maydis using cellular fractionation, atomic absorption spectrometry, and size exclusion chromatography with ICP-MS detection.

    Science.gov (United States)

    Muñoz, Alma Hortensia Serafin; Kubachka, Kevin; Wrobel, Kazimierz; Corona, Felix Gutierrez; Yathavakilla, Santha K V; Caruso, Joseph A; Wrobel, Katarzyna

    2005-06-29

    Huitlacoche is the ethnic name of the young fruiting bodies of Ustilago maydis, a common parasite of maize. In Mexico and other Latin American countries, this fungus has been traditionally appreciated as a local delicacy. In this work a metallomics approach was used with the determination of eight elements in huitlacoche by electrothermal atomic absorption spectrometry as one facet of this approach. The results obtained indicated relatively lower concentrations of commonly analyzed metals, as referred to the data reported for other mushroom types. This effect was ascribed to different accessibilities of elements, depending on fungus substrate (lower from plant than from soil). Subcellular fractionation was accomplished by centrifugation of cell homogenates suspended in Tris-HCl buffer. Recoveries of the fractionation procedure were in the range of 71-103%. For six elements (Cr, Cu, Fe, Mn, Ni, and Pb), the mean relative contributions in cytosol, cell walls, and mixed membrane fraction were 50.7, 48.2, and 1.1% respectively. To attain the molecular weight distribution of compounds containing target elements as an additional aspect of the metallomics approach, the fungus extract (1% sodium dodecyl sulfate in Tris-HCl, 30 mmol L(-)(1), pH 7.0) was analyzed by size exclusion chromatography with UV and ICP-MS detection. With spectrophotometric detection (280 nm), the elution of high molecular weight compounds was observed in the form of one peak (MW > 10 kDa), and several lower peaks appeared at higher retention times (MW < 10 kDa). On ICP-MS chromatograms, a coelution of (59)Co, (63)Cu, (57)Fe, (202)Hg, (60)Ni, and (80)Se with the first peak on the UV chromatogram was clearly observed, indicating that a fraction of each element incorporated with high molecular weight compounds (12.7, 19.8, 33.7, 100, 19.4, and 45.8%, respectively, based on the peak area measurements). From a comparison of (80)Se and (33)S chromatograms (for sulfur analysis, the extract was obtained in

  13. Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis.

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    Kämper, Jörg; Kahmann, Regine; Bölker, Michael; Ma, Li-Jun; Brefort, Thomas; Saville, Barry J; Banuett, Flora; Kronstad, James W; Gold, Scott E; Müller, Olaf; Perlin, Michael H; Wösten, Han A B; de Vries, Ronald; Ruiz-Herrera, José; Reynaga-Peña, Cristina G; Snetselaar, Karen; McCann, Michael; Pérez-Martín, José; Feldbrügge, Michael; Basse, Christoph W; Steinberg, Gero; Ibeas, Jose I; Holloman, William; Guzman, Plinio; Farman, Mark; Stajich, Jason E; Sentandreu, Rafael; González-Prieto, Juan M; Kennell, John C; Molina, Lazaro; Schirawski, Jan; Mendoza-Mendoza, Artemio; Greilinger, Doris; Münch, Karin; Rössel, Nicole; Scherer, Mario; Vranes, Miroslav; Ladendorf, Oliver; Vincon, Volker; Fuchs, Uta; Sandrock, Björn; Meng, Shaowu; Ho, Eric C H; Cahill, Matt J; Boyce, Kylie J; Klose, Jana; Klosterman, Steven J; Deelstra, Heine J; Ortiz-Castellanos, Lucila; Li, Weixi; Sanchez-Alonso, Patricia; Schreier, Peter H; Häuser-Hahn, Isolde; Vaupel, Martin; Koopmann, Edda; Friedrich, Gabi; Voss, Hartmut; Schlüter, Thomas; Margolis, Jonathan; Platt, Darren; Swimmer, Candace; Gnirke, Andreas; Chen, Feng; Vysotskaia, Valentina; Mannhaupt, Gertrud; Güldener, Ulrich; Münsterkötter, Martin; Haase, Dirk; Oesterheld, Matthias; Mewes, Hans-Werner; Mauceli, Evan W; DeCaprio, David; Wade, Claire M; Butler, Jonathan; Young, Sarah; Jaffe, David B; Calvo, Sarah; Nusbaum, Chad; Galagan, James; Birren, Bruce W

    2006-11-02

    Ustilago maydis is a ubiquitous pathogen of maize and a well-established model organism for the study of plant-microbe interactions. This basidiomycete fungus does not use aggressive virulence strategies to kill its host. U. maydis belongs to the group of biotrophic parasites (the smuts) that depend on living tissue for proliferation and development. Here we report the genome sequence for a member of this economically important group of biotrophic fungi. The 20.5-million-base U. maydis genome assembly contains 6,902 predicted protein-encoding genes and lacks pathogenicity signatures found in the genomes of aggressive pathogenic fungi, for example a battery of cell-wall-degrading enzymes. However, we detected unexpected genomic features responsible for the pathogenicity of this organism. Specifically, we found 12 clusters of genes encoding small secreted proteins with unknown function. A significant fraction of these genes exists in small gene families. Expression analysis showed that most of the genes contained in these clusters are regulated together and induced in infected tissue. Deletion of individual clusters altered the virulence of U. maydis in five cases, ranging from a complete lack of symptoms to hypervirulence. Despite years of research into the mechanism of pathogenicity in U. maydis, no 'true' virulence factors had been previously identified. Thus, the discovery of the secreted protein gene clusters and the functional demonstration of their decisive role in the infection process illuminate previously unknown mechanisms of pathogenicity operating in biotrophic fungi. Genomic analysis is, similarly, likely to open up new avenues for the discovery of virulence determinants in other pathogens.

  14. The pep4 gene encoding proteinase A is involved in dimorphism and pathogenesis of Ustilago maydis.

    Science.gov (United States)

    Soberanes-Gutiérrez, Cinthia V; Juárez-Montiel, Margarita; Olguín-Rodríguez, Omar; Hernández-Rodríguez, César; Ruiz-Herrera, José; Villa-Tanaca, Lourdes

    2015-10-01

    Vacuole proteases have important functions in different physiological processes in fungi. Taking this aspect into consideration, and as a continuation of our studies on the analysis of the proteolytic system of Ustilago maydis, a phytopathogenic member of the Basidiomycota, we have analysed the role of the pep4 gene encoding the vacuolar acid proteinase PrA in the pathogenesis and morphogenesis of the fungus. After confirmation of the location of the protease in the vacuole using fluorescent probes, we obtained deletion mutants of the gene in sexually compatible strains of U. maydis (FB1 and FB2), and analysed their phenotypes. It was observed that the yeast to mycelium dimorphic transition induced by a pH change in the medium, or the use of a fatty acid as sole carbon source, was severely reduced in Δpep4 mutants. In addition, the virulence of the mutants in maize seedlings was reduced, as revealed by the lower proportion of plants infected and the reduction in size of the tumours induced by the pathogen, when compared with wild-type strains. All of these phenotypic alterations were reversed by complementation of the mutant strains with the wild-type gene. These results provide evidence of the importance of the pep4 gene for the morphogenesis and virulence of U. maydis.

  15. A novel intracellular nitrogen-fixing symbiosis made by Ustilago maydis and Bacillus spp.

    Science.gov (United States)

    Ruiz-Herrera, José; León-Ramírez, Claudia; Vera-Nuñez, Antonio; Sánchez-Arreguín, Alejandro; Ruiz-Medrano, Roberto; Salgado-Lugo, Holjes; Sánchez-Segura, Lino; Peña-Cabriales, Juan José

    2015-08-01

    We observed that the maize pathogenic fungus Ustilago maydis grew in nitrogen (N)-free media at a rate similar to that observed in media containing ammonium nitrate, suggesting that it was able to fix atmospheric N2 . Because only prokaryotic organisms have the capacity to reduce N2 , we entertained the possibility that U. maydis was associated with an intracellular bacterium. The presence of nitrogenase in the fungus was analyzed by acetylene reduction, and capacity to fix N2 by use of (15) N2 . Presence of an intracellular N2 -fixing bacterium was analyzed by PCR amplification of bacterial 16S rRNA and nifH genes, and by microscopic observations. Nitrogenase activity and (15) N incorporation into the cells proved that U. maydis fixed N2 . Light and electron microscopy, and fluorescence in situ hybridization (FISH) experiments revealed the presence of intracellular bacteria related to Bacillus pumilus, as evidenced by sequencing of the PCR-amplified fragments. These observations reveal for the first time the existence of an endosymbiotic N2 -fixing association involving a fungus and a bacterium.

  16. Two members of the Ustilago maydis velvet family influence teliospore development and virulence on maize seedlings.

    Science.gov (United States)

    Karakkat, Brijesh B; Gold, Scott E; Covert, Sarah F

    2013-12-01

    Members of the fungal-specific velvet protein family regulate sexual and asexual spore production in the Ascomycota. We predicted, therefore, that velvet homologs in the basidiomycetous plant pathogen Ustilago maydis would regulate sexual spore development, which is also associated with plant disease progression in this fungus. To test this hypothesis, we studied the function of three U. maydis velvet genes, umv1, umv2 and umv3. Using a gene replacement strategy, deletion mutants were made in all three genes in compatible haploid strains, and additionally for umv1 and umv2 in the solopathogenic strain, SG200. None of the mutants showed novel morphological phenotypes during yeast-like, in vitro growth. However, the Δumv1 mutants failed to induce galls or teliospores in maize. Chlorazol black E staining of leaves infected with Δumv1 dikaryons revealed that the Δumv1 hyphae did not proliferate normally and were blocked developmentally before teliospore formation. The Δumv2 mutants were able to induce galls and teliospores in maize, but were slow to do so and thus reduced in virulence. The Δumv3 mutants were not affected in teliospore formation or disease progression. Complementation of the Δumv1 and Δumv2 mutations in the SG200 background produced disease indices similar to those of SG200. These results indicate that two U. maydis velvet family members, umv1 and umv2, are important for normal teliospore development and disease progression in maize seedlings.

  17. The DNA damage response signaling cascade regulates proliferation of the phytopathogenic fungus Ustilago maydis in planta.

    Science.gov (United States)

    de Sena-Tomás, Carmen; Fernández-Álvarez, Alfonso; Holloman, William K; Pérez-Martín, José

    2011-04-01

    In the phytopathogenic fungus Ustilago maydis, the dikaryotic state dominates the period of growth occurring during the infectious phase. Dikaryons are cells in which two nuclei, one from each parent cell, share a single cytoplasm for a period of time without undergoing nuclear fusion. In fungal cells, maintenance of the dikaryotic state requires an intricate cell division process that often involves the formation of a structure known as the clamp connection as well as the sorting of one of the nuclei to this structure to ensure that each daughter dikaryon inherits a balance of each parental genome. Here, we describe an atypical role of the DNA damage checkpoint kinases Chk1 and Atr1 during pathogenic growth of U. maydis. We found that Chk1 and Atr1 collaborate to control cell cycle arrest during the induction of the virulence program in U. maydis and that Chk1 and Atr1 work together to control the dikaryon formation. These findings uncover a link between a widely conserved signaling cascade and the virulence program in a phytopathogen. We propose a model in which adjustment of the cell cycle by the Atr1-Chk1 axis controls fidelity in dikaryon formation. Therefore, Chk1 and Atr1 emerge as critical cell type regulators in addition to their roles in the DNA damage response.

  18. Defects in mitochondrial and peroxisomal β-oxidation influence virulence in the maize pathogen Ustilago maydis.

    Science.gov (United States)

    Kretschmer, Matthias; Klose, Jana; Kronstad, James W

    2012-08-01

    An understanding of metabolic adaptation during the colonization of plants by phytopathogenic fungi is critical for developing strategies to protect crops. Lipids are abundant in plant tissues, and fungal phytopathogens in the phylum basidiomycota possess both peroxisomal and mitochondrial β-oxidation pathways to utilize this potential carbon source. Previously, we demonstrated a role for the peroxisomal β-oxidation enzyme Mfe2 in the filamentous growth, virulence, and sporulation of the maize pathogen Ustilago maydis. However, mfe2 mutants still caused disease symptoms, thus prompting a more detailed investigation of β-oxidation. We now demonstrate that a defect in the had1 gene encoding hydroxyacyl coenzyme A dehydrogenase for mitochondrial β-oxidation also influences virulence, although its paralog, had2, makes only a minor contribution. Additionally, we identified a gene encoding a polypeptide with similarity to the C terminus of Mfe2 and designated it Mfe2b; this gene makes a contribution to virulence only in the background of an mfe2Δ mutant. We also show that short-chain fatty acids induce cell death in U. maydis and that a block in β-oxidation leads to toxicity, likely because of the accumulation of toxic intermediates. Overall, this study reveals that β-oxidation has a complex influence on the formation of disease symptoms by U. maydis that includes potential metabolic contributions to proliferation in planta and an effect on virulence-related morphogenesis.

  19. Conserved and distinct functions of the "stunted" (StuA)-Homolog Ust1 during cell differentiation in the corn smut fungus Ustilago maydis

    Science.gov (United States)

    Ustilago maydis, causal agent of corn smut, is a model for obligate fungal plant pathogens because, although it can proliferate saprobically in its yeast form, the infectious filamentous form is absolutely dependent on the host to complete its life cycle. Maize responds to U. maydis colonization by...

  20. Ustilago maydis phosphodiesterases play a role in the dimorphic switch and in pathogenicity.

    Science.gov (United States)

    Agarwal, Charu; Aulakh, Kavita B; Edelen, Kaly; Cooper, Michael; Wallen, R Margaret; Adams, Seth; Schultz, David J; Perlin, Michael H

    2013-05-01

    Components of the cAMP (cyclic AMP) signalling cascades are conserved from fungi to humans, and are particularly important for fungal dimorphism and pathogenicity. Previous work has described two phosphodiesterases, UmPde1 and UmPde2, in Ustilago maydis which show strong phosphodiesterase activity. We further characterized the biological function(s) of these phosphodiesterases in U. maydis. Specifically, we examined their possible role(s) in regulation of the cAMP-dependent protein kinase A (PKA) pathway and their roles in filamentous growth and pathogenicity. We found that UmPde1, which shares 35 % similarity with Cryptococcus neoformans Pde1, also displays functional homology with this enzyme. UmPde1 complements the capsule-formation defect of C. neoformans strains deleted for Pde1. In U. maydis, the cell morphology of the umpde1 deletion mutant resembled the multiple budding phenotypes seen with the ubc1 mutant, which lacks the regulatory subunit of PKA. Interestingly, on low-ammonium medium, umpde2 deletion strains showed a reduction in filamentation that was comparable to that of ubc1 deletion strains; however, umpde1 deletion strains showed normal filamentation on low-ammonium medium. Furthermore, both the ubc1 deletion strain in which the PKA pathway was constitutively active and the umpde1 deletion strains were significantly reduced in pathogenicity, while the umpde2 deletion strains showed a trend for reduced pathogenicity compared with wild-type strains. These data support a role for the phosphodiesterases UmPde1 and UmPde2 in regulating the U. maydis cAMP-dependent PKA pathway through modulation of cAMP levels, thus affecting dimorphic growth and pathogenicity.

  1. In vitro interactions between Fusarium verticillioides and Ustilago maydis through real-time PCR and metabolic profiling.

    Science.gov (United States)

    Rodriguez Estrada, Alma E; Hegeman, Adrian; Kistler, H Corby; May, Georgiana

    2011-09-01

    The goal of this research was to determine mechanisms of interaction between endophytic strains of Fusarium verticillioides (Sacc.) Nirenberg and the pathogen, Ustilago maydis (DC) (Corda). Endophytic strains of the fungus F. verticillioides are commonly found in association with maize (Zea mays) and when co-inoculated with U. maydis, often lead to decreased disease severity caused by the pathogen. Here, we developed methods (liquid chromatography-mass spectrometry) to evaluate changes in relative concentration of metabolites produced during in vitro interactions between the endophyte and pathogen. Fungi were grown on two different media, in single and in confronted cultures. We used real-time PCR (qPCR) assays to measure relative changes in fungal biomass, that occurred in confronted cultures compared to single cultures. The results showed that most secondary metabolites are constitutively produced by each species. Metabolite profiles are complex for U. maydis (twenty chromatographic peaks detected) while relatively fewer compounds were detected for F. verticillioides (six chromatographic peaks). In confronted cultures, metabolite ratio (metabolite concentration/biomass) generally increases for U. maydis metabolites while no significant changes were observed for most F. verticillioides metabolites. The results show that F. verticillioides is a strong antagonist of U. maydis as its presence leads to large reductions in U. maydis biomass. We infer that few U. maydis metabolites likely serve antibiotic functions against F. verticillioides. The methods described here are sufficiently sensitive to detect small changes in biomass and metabolite concentration associated with differing genotypes of the interacting species.

  2. Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis.

    Science.gov (United States)

    van der Linde, Karina; Kastner, Christine; Kumlehn, Jochen; Kahmann, Regine; Doehlemann, Gunther

    2011-01-01

    Infection of maize (Zea mays) plants with the corn smut fungus Ustilago maydis leads to the formation of large tumors on the stem, leaves and inflorescences. In this biotrophic interaction, plant defense responses are actively suppressed by the pathogen, and previous transcriptome analyses of infected maize plants showed massive and stage-specific changes in host gene expression during disease progression. To identify maize genes that are functionally involved in the interaction with U. maydis, we adapted a virus-induced gene silencing (VIGS) system based on the brome mosaic virus (BMV) for maize. Conditions were established that allowed successful U. maydis infection of BMV-preinfected maize plants. This set-up enabled quantification of VIGS and its impact on U. maydis infection using a quantitative real-time PCR (qRT-PCR)-based readout. In proof-of-principle experiments, an U. maydis-induced terpene synthase was shown to negatively regulate disease development while a protein involved in cell death inhibition was required for full virulence of U. maydis. The results suggest that this system is a versatile tool for the rapid identification of maize genes that determine compatibility with U. maydis.

  3. The telomerase reverse transcriptase subunit from the dimorphic fungus Ustilago maydis.

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    Dolores Bautista-España

    Full Text Available In this study, we investigated the reverse transcriptase subunit of telomerase in the dimorphic fungus Ustilago maydis. This protein (Trt1 contains 1371 amino acids and all of the characteristic TERT motifs. Mutants created by disrupting trt1 had senescent traits, such as delayed growth, low replicative potential, and reduced survival, that were reminiscent of the traits observed in est2 budding yeast mutants. Telomerase activity was observed in wild-type fungus sporidia but not those of the disruption mutant. The introduction of a self-replicating plasmid expressing Trt1 into the mutant strain restored growth proficiency and replicative potential. Analyses of trt1 crosses in planta suggested that Trt1 is necessary for teliospore formation in homozygous disrupted diploids and that telomerase is haploinsufficient in heterozygous diploids. Additionally, terminal restriction fragment analysis in the progeny hinted at alternative survival mechanisms similar to those of budding yeast.

  4. Characterization and high expression of recombinant Ustilago maydis xylanase in Pichia pastoris.

    Science.gov (United States)

    Han, Hongjuan; You, Shuang; Zhu, Bo; Fu, Xiaoyan; Sun, Baihui; Qiu, Jin; Yu, Chengye; Chen, Lei; Peng, Rihe; Yao, Quanhong

    2015-03-01

    A recombinant xylanase gene (rxynUMB) from Ustilago maydis 521 was expressed in Pichia pastoris, and the enzyme was purified and characterized. Phylogenetic analysis demonstrated that rxynUMB belongs to glycosyl hydrolase family 11. The Trp84, Trp95, Glu93, and Glu189 residues are proposed to be present at the active site. The apparent molecular mass of the recombinant xylananse was approximately 24 kDa, and the optimum pH and temperature were 4.3 and 50 °C, respectively. Xylanase activity was enhanced by 166 and 115% with Fe(2+) and Mn(2+), respectively. The biochemical properties of this recombinant xylanase suggest that it may be a useful candidate for a variety of commercial applications.

  5. The telomerase reverse transcriptase subunit from the dimorphic fungus Ustilago maydis.

    Science.gov (United States)

    Bautista-España, Dolores; Anastacio-Marcelino, Estela; Horta-Valerdi, Guillermo; Celestino-Montes, Antonio; Kojic, Milorad; Negrete-Abascal, Erasmo; Reyes-Cervantes, Hortensia; Vázquez-Cruz, Candelario; Guzmán, Plinio; Sánchez-Alonso, Patricia

    2014-01-01

    In this study, we investigated the reverse transcriptase subunit of telomerase in the dimorphic fungus Ustilago maydis. This protein (Trt1) contains 1371 amino acids and all of the characteristic TERT motifs. Mutants created by disrupting trt1 had senescent traits, such as delayed growth, low replicative potential, and reduced survival, that were reminiscent of the traits observed in est2 budding yeast mutants. Telomerase activity was observed in wild-type fungus sporidia but not those of the disruption mutant. The introduction of a self-replicating plasmid expressing Trt1 into the mutant strain restored growth proficiency and replicative potential. Analyses of trt1 crosses in planta suggested that Trt1 is necessary for teliospore formation in homozygous disrupted diploids and that telomerase is haploinsufficient in heterozygous diploids. Additionally, terminal restriction fragment analysis in the progeny hinted at alternative survival mechanisms similar to those of budding yeast.

  6. Endoplasmic Reticulum Glucosidase II Is Required for Pathogenicity of Ustilago maydisW⃞

    Science.gov (United States)

    Schirawski, Jan; Böhnert, Heidi U.; Steinberg, Gero; Snetselaar, Karen; Adamikowa, Lubica; Kahmann, Regine

    2005-01-01

    We identified a nonpathogenic strain of Ustilago maydis by tagging mutagenesis. The affected gene, glucosidase1 (gas1), displays similarity to catalytic α-subunits of endoplasmic reticulum (ER) glucosidase II. We have shown that Gas1 localizes to the ER and complements the temperature-sensitive phenotype of a Saccharomyces cerevisiae mutant lacking ER glucosidase II. gas1 deletion mutants were normal in growth and mating but were more sensitive to calcofluor and tunicamycin. Mutant infection hyphae displayed significant alterations in the distribution of cell wall material and were able to form appressoria and penetrate the plant surface but arrested growth in the epidermal cell layer. Electron microscopy analysis revealed that the plant–fungal interface between mutant hyphae and the plant plasma membrane was altered compared with the interface of penetrating wild-type hyphae. This may indicate that gas1 mutants provoke a plant response. PMID:16272431

  7. Endoplasmic reticulum glucosidase II is required for pathogenicity of Ustilago maydis.

    Science.gov (United States)

    Schirawski, Jan; Böhnert, Heidi U; Steinberg, Gero; Snetselaar, Karen; Adamikowa, Lubica; Kahmann, Regine

    2005-12-01

    We identified a nonpathogenic strain of Ustilago maydis by tagging mutagenesis. The affected gene, glucosidase1 (gas1), displays similarity to catalytic alpha-subunits of endoplasmic reticulum (ER) glucosidase II. We have shown that Gas1 localizes to the ER and complements the temperature-sensitive phenotype of a Saccharomyces cerevisiae mutant lacking ER glucosidase II. gas1 deletion mutants were normal in growth and mating but were more sensitive to calcofluor and tunicamycin. Mutant infection hyphae displayed significant alterations in the distribution of cell wall material and were able to form appressoria and penetrate the plant surface but arrested growth in the epidermal cell layer. Electron microscopy analysis revealed that the plant-fungal interface between mutant hyphae and the plant plasma membrane was altered compared with the interface of penetrating wild-type hyphae. This may indicate that gas1 mutants provoke a plant response.

  8. Huitlacoche yield in some maize varieties in the Mediterranean region of Turkey

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    Mehmet Aydoğdu

    2015-06-01

    Full Text Available Huitlacoche is the Aztecs name given to the smut galls on ears of maize caused by the pathogenic plant fungus Ustilago maydis [(DC Corda.]. It is known as maize mushroom, and it has been considered a delicacy and in Mesoamerica. The aim of the present study was to determine the responses of some maize varieties to the growth of the fungus in order to evaluate the prospect production of these smutty ears as a maize mushroom. A 2-year study was conducted in the Mediterranean region of Turkey in 2010 and 2011. Inoculations were performed by injecting inoculum into the ear through the silk channel of plants in plots. Each treatment had control plots. Average ear-gall (huitlacoche severity and incidence of all the varieties were at the rates of 4.0 and 41.6%, respectively. However, the highest severity of ear-gall (6.5 and incidence (60.6% were found in Karadeniz Yıldızı flint maize variety; colossal smutty ears were formed in the maize cultivars. This study showed that certain maize cultivars (flint corn and dent corn can be used efficiently in the production of huitlacoche.

  9. Hxt1, a monosaccharide transporter and sensor required for virulence of the maize pathogen Ustilago maydis.

    Science.gov (United States)

    Schuler, David; Wahl, Ramon; Wippel, Kathrin; Vranes, Miroslav; Münsterkötter, Martin; Sauer, Norbert; Kämper, Jörg

    2015-05-01

    The smut Ustilago maydis, a ubiquitous pest of corn, is highly adapted to its host to parasitize on its organic carbon sources. We have identified a hexose transporter, Hxt1, as important for fungal development during both the saprophytic and the pathogenic stage of the fungus. Hxt1 was characterized as a high-affinity transporter for glucose, fructose, and mannose; ∆hxt1 strains show significantly reduced growth on these substrates, setting Hxt1 as the main hexose transporter during saprophytic growth. After plant infection, ∆hxt1 strains show decreased symptom development. However, expression of a Hxt1 protein with a mutation leading to constitutively active signaling in the yeast glucose sensors Snf3p and Rgt2p results in completely apathogenic strains. Fungal development is stalled immediately after plant penetration, implying a dual function of Hxt1 as transporter and sensor. As glucose sensors are only known for yeasts, 'transceptor' as Hxt1 may constitute a general mechanism for sensing of glucose in fungi. In U. maydis, Hxt1 links a nutrient-dependent environmental signal to the developmental program during pathogenic development.

  10. The Ustilago maydis effector Pep1 suppresses plant immunity by inhibition of host peroxidase activity.

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    Christoph Hemetsberger

    Full Text Available The corn smut Ustilago maydis establishes a biotrophic interaction with its host plant maize. This interaction requires efficient suppression of plant immune responses, which is attributed to secreted effector proteins. Previously we identified Pep1 (Protein essential during penetration-1 as a secreted effector with an essential role for U. maydis virulence. pep1 deletion mutants induce strong defense responses leading to an early block in pathogenic development of the fungus. Using cytological and functional assays we show that Pep1 functions as an inhibitor of plant peroxidases. At sites of Δpep1 mutant penetrations, H₂O₂ strongly accumulated in the cell walls, coinciding with a transcriptional induction of the secreted maize peroxidase POX12. Pep1 protein effectively inhibited the peroxidase driven oxidative burst and thereby suppresses the early immune responses of maize. Moreover, Pep1 directly inhibits peroxidases in vitro in a concentration-dependent manner. Using fluorescence complementation assays, we observed a direct interaction of Pep1 and the maize peroxidase POX12 in vivo. Functional relevance of this interaction was demonstrated by partial complementation of the Δpep1 mutant defect by virus induced gene silencing of maize POX12. We conclude that Pep1 acts as a potent suppressor of early plant defenses by inhibition of peroxidase activity. Thus, it represents a novel strategy for establishing a biotrophic interaction.

  11. A secreted Ustilago maydis effector promotes virulence by targeting anthocyanin biosynthesis in maize.

    Science.gov (United States)

    Tanaka, Shigeyuki; Brefort, Thomas; Neidig, Nina; Djamei, Armin; Kahnt, Jörg; Vermerris, Wilfred; Koenig, Stefanie; Feussner, Kirstin; Feussner, Ivo; Kahmann, Regine

    2014-01-01

    The biotrophic fungus Ustilago maydis causes smut disease in maize with characteristic tumor formation and anthocyanin induction. Here, we show that anthocyanin biosynthesis is induced by the virulence promoting secreted effector protein Tin2. Tin2 protein functions inside plant cells where it interacts with maize protein kinase ZmTTK1. Tin2 masks a ubiquitin-proteasome degradation motif in ZmTTK1, thus stabilizing the active kinase. Active ZmTTK1 controls activation of genes in the anthocyanin biosynthesis pathway. Without Tin2, enhanced lignin biosynthesis is observed in infected tissue and vascular bundles show strong lignification. This is presumably limiting access of fungal hyphae to nutrients needed for massive proliferation. Consistent with this assertion, we observe that maize brown midrib mutants affected in lignin biosynthesis are hypersensitive to U. maydis infection. We speculate that Tin2 rewires metabolites into the anthocyanin pathway to lower their availability for other defense responses. DOI: http://dx.doi.org/10.7554/eLife.01355.001.

  12. Programmed cell cycle arrest is required for infection of corn plants by the fungus Ustilago maydis.

    Science.gov (United States)

    Castanheira, Sónia; Mielnichuk, Natalia; Pérez-Martín, José

    2014-12-01

    Ustilago maydis is a plant pathogen that requires a specific structure called infective filament to penetrate the plant tissue. Although able to grow, this filament is cell cycle arrested on the plant surface. This cell cycle arrest is released once the filament penetrates the plant tissue. The reasons and mechanisms for this cell cycle arrest are unknown. Here, we have tried to address these questions. We reached three conclusions from our studies. First, the observed cell cycle arrest is the result of the cooperation of at least two distinct mechanisms: one involving the activation of the DNA damage response (DDR) cascade; and the other relying on the transcriptional downregulation of Hsl1, a kinase that modulates the G2/M transition. Second, a sustained cell cycle arrest during the infective filament step is necessary for the virulence in U. maydis, as a strain unable to arrest the cell cycle was severely impaired in its ability to infect corn plants. Third, production of the appressorium, a structure required for plant penetration, is incompatible with an active cell cycle. The inability to infect plants by strains defective in cell cycle arrest seems to be caused by their failure to induce the appressorium formation process. In summary, our findings uncover genetic circuits to arrest the cell cycle during the growth of this fungus on the plant surface, thus allowing the penetration into plant tissue.

  13. The Ustilago maydis effector Pep1 suppresses plant immunity by inhibition of host peroxidase activity.

    Science.gov (United States)

    Hemetsberger, Christoph; Herrberger, Christian; Zechmann, Bernd; Hillmer, Morten; Doehlemann, Gunther

    2012-01-01

    The corn smut Ustilago maydis establishes a biotrophic interaction with its host plant maize. This interaction requires efficient suppression of plant immune responses, which is attributed to secreted effector proteins. Previously we identified Pep1 (Protein essential during penetration-1) as a secreted effector with an essential role for U. maydis virulence. pep1 deletion mutants induce strong defense responses leading to an early block in pathogenic development of the fungus. Using cytological and functional assays we show that Pep1 functions as an inhibitor of plant peroxidases. At sites of Δpep1 mutant penetrations, H₂O₂ strongly accumulated in the cell walls, coinciding with a transcriptional induction of the secreted maize peroxidase POX12. Pep1 protein effectively inhibited the peroxidase driven oxidative burst and thereby suppresses the early immune responses of maize. Moreover, Pep1 directly inhibits peroxidases in vitro in a concentration-dependent manner. Using fluorescence complementation assays, we observed a direct interaction of Pep1 and the maize peroxidase POX12 in vivo. Functional relevance of this interaction was demonstrated by partial complementation of the Δpep1 mutant defect by virus induced gene silencing of maize POX12. We conclude that Pep1 acts as a potent suppressor of early plant defenses by inhibition of peroxidase activity. Thus, it represents a novel strategy for establishing a biotrophic interaction.

  14. Endoplasmic reticulum glucosidases and protein quality control factors cooperate to establish biotrophy in Ustilago maydis.

    Science.gov (United States)

    Fernández-Álvarez, Alfonso; Elías-Villalobos, Alberto; Jiménez-Martín, Alberto; Marín-Menguiano, Miriam; Ibeas, José I

    2013-11-01

    Secreted fungal effectors mediate plant-fungus pathogenic interactions. These proteins are typically N-glycosylated, a common posttranslational modification affecting their location and function. N-glycosylation consists of the addition, and subsequent maturation, of an oligosaccharide core in the endoplasmic reticulum (ER) and Golgi apparatus. In this article, we show that two enzymes catalyzing specific stages of this pathway in maize smut (Ustilago maydis), glucosidase I (Gls1) and glucosidase II β-subunit (Gas2), are essential for its pathogenic interaction with maize (Zea mays). Gls1 is required for the initial stages of infection following appressorium penetration, and Gas2 is required for efficient fungal spreading inside infected tissues. While U. maydis Δgls1 cells induce strong plant defense responses, Δgas2 hyphae are able to repress them, showing that slight differences in the N-glycoprotein processing can determine the extent of plant-fungus interactions. Interestingly, the calnexin protein, a central element of the ER quality control system for N-glycoproteins in eukaryotic cells, is essential for avoiding plant defense responses in cells with defective N-glycoproteins processing. Thus, N-glycoprotein maturation and this conserved checkpoint appear to play an important role in the establishment of an initial biotrophic state with the plant, which allows subsequent colonization.

  15. Crosstalk between the unfolded protein response and pathways that regulate pathogenic development in Ustilago maydis.

    Science.gov (United States)

    Heimel, Kai; Freitag, Johannes; Hampel, Martin; Ast, Julia; Bölker, Michael; Kämper, Jörg

    2013-10-01

    The unfolded protein response (UPR) is a conserved eukaryotic signaling pathway regulating endoplasmic reticulum (ER) homeostasis during ER stress, which results, for example, from an increased demand for protein secretion. Here, we characterize the homologs of the central UPR regulatory proteins Hac1 (for Homologous to ATF/CREB1) and Inositol Requiring Enzyme1 in the plant pathogenic fungus Ustilago maydis and demonstrate that the UPR is tightly interlinked with the b mating-type-dependent signaling pathway that regulates pathogenic development. Exact timing of UPR is required for virulence, since premature activation interferes with the b-dependent switch from budding to filamentous growth. In addition, we found crosstalk between UPR and the b target Clampless1 (Clp1), which is essential for cell cycle release and proliferation in planta. The unusual C-terminal extension of the U. maydis Hac1 homolog, Cib1 (for Clp1 interacting bZIP1), mediates direct interaction with Clp1. The interaction between Clp1 and Cib1 promotes stabilization of Clp1, resulting in enhanced ER stress tolerance that prevents deleterious UPR hyperactivation. Thus, the interaction between Cib1 and Clp1 constitutes a checkpoint to time developmental progression and increased secretion of effector proteins at the onset of biotrophic development. Crosstalk between UPR and the b mating-type regulated developmental program adapts ER homeostasis to the changing demands during biotrophy.

  16. Absence of repellents in Ustilago maydis induces genes encoding small secreted proteins.

    Science.gov (United States)

    Teertstra, Wieke R; Krijgsheld, Pauline; Wösten, Han A B

    2011-08-01

    The rep1 gene of the maize pathogen Ustilago maydis encodes a pre-pro-protein that is processed in the secretory pathway into 11 peptides. These so-called repellents form amphipathic amyloid fibrils at the surface of aerial hyphae. A SG200 strain in which the rep1 gene is inactivated (∆rep1 strain) is affected in aerial hyphae formation. We here assessed changes in global gene expression as a consequence of the inactivation of the rep1 gene. Microarray analysis revealed that only 31 genes in the ∆rep1 SG200 strain had a fold change in expression of ≥2. Twenty-two of these genes were up-regulated and half of them encode small secreted proteins (SSPs) with unknown functions. Seven of the SSP genes and two other genes that are over-expressed in the ∆rep1 SG200 strain encode proteins that can be classified as secreted cysteine-rich proteins (SCRPs). Interestingly, most of the SCRPs are predicted to form amyloids. The SCRP gene um00792 showed the highest up-regulation in the ∆rep1 strain. Using GFP as a reporter, it was shown that this gene is over-expressed in the layer of hyphae at the medium-air interface. Taken together, it is concluded that inactivation of rep1 hardly affects the expression profile of U. maydis, despite the fact that the mutant strain has a strong reduced ability to form aerial hyphae.

  17. Phenotypic comparison of samdc and spe mutants reveals complex relationships of polyamine metabolism in Ustilago maydis.

    Science.gov (United States)

    Valdés-Santiago, Laura; Cervantes-Chávez, José Antonio; Winkler, Robert; León-Ramírez, Claudia G; Ruiz-Herrera, José

    2012-03-01

    Synthesis of spermidine involves the action of two enzymes, spermidine synthase (Spe) and S-adenosylmethionine decarboxylase (Samdc). Previously we cloned and disrupted the gene encoding Spe as a first approach to unravel the biological function of spermidine in Ustilago maydis. With this background, the present study was designed to provide a better understanding of the role played by Samdc in the regulation of the synthesis of this polyamine. With this aim we proceeded to isolate and delete the gene encoding Samdc from U. maydis, and made a comparative analysis of the phenotypes of samdc and spe mutants. Both spe and samdc mutants behaved as spermidine auxotrophs, and were more sensitive than the wild-type strain to different stress conditions. However, the two mutants displayed significant differences: in contrast to spe mutants, samdc mutants were more sensitive to LiCl stress, high spermidine concentrations counteracted their dimorphic deficiency, and they were completely avirulent. It is suggested that these differences are possibly related to differences in exogenous spermidine uptake or the differential location of the respective enzymes in the cell. Alternatively, since samdc mutants accumulate higher levels of S-adenosylmethionine (SAM), whereas spe mutants accumulate decarboxylated SAM, the known opposite roles of these metabolites in the processes of methylation and differentiation offer an additional attractive hypothesis to explain the phenotypic differences of the two mutants, and provide insights into the additional roles of polyamine metabolism in the physiology of the cell.

  18. Insights into Host Cell Modulation and Induction of New Cells by the Corn Smut Ustilago maydis

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    Amey Redkar

    2017-05-01

    Full Text Available Many filamentous fungal pathogens induce drastic modulation of host cells causing abnormal infectious structures such as galls, or tumors that arise as a result of re-programming in the original developmental cell fate of a colonized host cell. Developmental consequences occur predominantly with biotrophic phytopathogens. This suggests that these host structures result as an outcome of efficient defense suppression and intimate fungal–host interaction to suit the pathogen’s needs for completion of its infection cycle. This mini-review mainly summarizes host cell re-programming that occurs in the Ustilago maydis – maize interaction, in which the pathogen deploys cell-type specific effector proteins with varying activities. The fungus senses the physiological status and identity of colonized host cells and re-directs the endogenous developmental program of its host. The disturbance of host cell physiology and cell fate leads to novel cell shapes, increased cell size, and/or the number of host cells. We particularly highlight the strategies of U. maydis to induce physiologically varied host organs to form the characteristic tumors in both vegetative and floral parts of maize.

  19. A ferroxidation/permeation iron uptake system is required for virulence in Ustilago maydis.

    Science.gov (United States)

    Eichhorn, Heiko; Lessing, Franziska; Winterberg, Britta; Schirawski, Jan; Kämper, Jörg; Müller, Philip; Kahmann, Regine

    2006-11-01

    In the smut fungus Ustilago maydis, a tightly regulated cAMP signaling cascade is necessary for pathogenic development. Transcriptome analysis using whole genome microarrays set up to identify putative target genes of the protein kinase A catalytic subunit Adr1 revealed nine genes with putative functions in two high-affinity iron uptake systems. These genes locate to three gene clusters on different chromosomes and include the previously identified complementing siderophore auxotroph genes sid1 and sid2 involved in siderophore biosynthesis. Transcription of all nine genes plus three additional genes associated with the gene clusters was also coregulated by iron through the Urbs1 transcription factor. Two components of a high-affinity iron uptake system were characterized in more detail: fer2, encoding a high-affinity iron permease; and fer1, encoding an iron multicopper oxidase. Fer2 localized to the plasma membrane and complemented an ftr1 mutant of Saccharomyces cerevisiae lacking a high-affinity iron permease. During pathogenic development, fer2 expression was confined to the phase of hyphal proliferation inside the plant. fer2 as well as fer1 deletion mutants were strongly affected in virulence. These data highlight the importance of the high-affinity iron uptake system via an iron permease and a multicopper oxidase for biotrophic development in the U. maydis/maize (Zea mays) pathosystem.

  20. A Ferroxidation/Permeation Iron Uptake System Is Required for Virulence in Ustilago maydis[W

    Science.gov (United States)

    Eichhorn, Heiko; Lessing, Franziska; Winterberg, Britta; Schirawski, Jan; Kämper, Jörg; Müller, Philip; Kahmann, Regine

    2006-01-01

    In the smut fungus Ustilago maydis, a tightly regulated cAMP signaling cascade is necessary for pathogenic development. Transcriptome analysis using whole genome microarrays set up to identify putative target genes of the protein kinase A catalytic subunit Adr1 revealed nine genes with putative functions in two high-affinity iron uptake systems. These genes locate to three gene clusters on different chromosomes and include the previously identified complementing siderophore auxotroph genes sid1 and sid2 involved in siderophore biosynthesis. Transcription of all nine genes plus three additional genes associated with the gene clusters was also coregulated by iron through the Urbs1 transcription factor. Two components of a high-affinity iron uptake system were characterized in more detail: fer2, encoding a high-affinity iron permease; and fer1, encoding an iron multicopper oxidase. Fer2 localized to the plasma membrane and complemented an ftr1 mutant of Saccharomyces cerevisiae lacking a high-affinity iron permease. During pathogenic development, fer2 expression was confined to the phase of hyphal proliferation inside the plant. fer2 as well as fer1 deletion mutants were strongly affected in virulence. These data highlight the importance of the high-affinity iron uptake system via an iron permease and a multicopper oxidase for biotrophic development in the U. maydis/maize (Zea mays) pathosystem. PMID:17138696

  1. Two phosphodiesterases from Ustilago maydis share structural and biochemical properties with non-fungal phosphodiesterases

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    Charu eAgarwal

    2010-11-01

    Full Text Available The dependence of Protein Kinase A (PKA activity on cAMP levels is an important facet of the dimorphic switch between budding and filamentous growth as well as for pathogenicity in some fungi. To better understand these processes in the pathogenic fungus Ustilago maydis, we characterized the structure and biochemical functions of two phosphodiesterase (PDE genes. Phosphodiesterases are enzymes involved in cAMP turnover and thus, contribute to the regulation of the cAMP-PKA signaling pathway. Two predicted homologues of PDEs were identified in the genome of U. maydis and hypothesized to be involved in cAMP turnover, thus regulating activity of the PKA catalytic subunit. Both umpde1 and umpde2 genes contain domains associated with phosphodiesterase activity predicted by InterPro analysis. Biochemical characterization of recombinantly produced UmPde1 (U. maydis Phosphodiesterase I and UmPde2 demonstrated that both enzymes have phosphodiesterase activity in vitro, yet neither was inhibited by the phosphodiesterase inhibitor IBMX. Moreover, UmPde1 is specific for cAMP, while UmPde2 has broader substrate specificity, utilizing cAMP and cGMP as substrates. In addition, UmPde2 was also found to have nucleotide phosphatase activity that was higher with GMP compared to AMP. These results demonstrate that UmPde1 is a bona fide phosphodiesterase, while UmPde2 has more general activity as a cyclic nucleotide phosphodiesterase and/or GMP/AMP phosphatase. Thus, UmPde1 and UmPde2 likely have important roles in cell morphology and development and share some characteristics with a variety of non-fungal phosphodiesterases.

  2. Isolation of UmRrm75, a gene involved in dimorphism and virulence of Ustilago maydis.

    Science.gov (United States)

    Rodríguez-Kessler, Margarita; Baeza-Montañez, Lourdes; García-Pedrajas, María D; Tapia-Moreno, Alejandro; Gold, Scott; Jiménez-Bremont, Juan F; Ruiz-Herrera, José

    2012-05-20

    Ustilago maydis displays dimorphic growth, alternating between a saprophytic haploid yeast form and a filamentous dikaryon, generated by mating of haploid cells and which is an obligate parasite. Induction of the dimorphic transition of haploid strains in vitro by change in ambient pH has been used to understand the mechanisms governing this differentiation process. In this study we used suppression subtractive hybridization to generate a cDNA library of U. maydis genes up-regulated in the filamentous form induced in vitro at acid pH. Expression analysis using quantitative RT-PCR showed that the induction of two unigenes identified in this library coincided with the establishment of filamentous growth in the acid pH medium. This expression pattern suggested that they were specifically associated to hyphal development rather than merely acid pH-induced genes. One of these genes, UmRrm75, encodes a protein containing three RNA recognition motifs and glycine-rich repeats and was selected for further study. The UmRrm75 gene contains 4 introns, and produces a splicing variant by a 3'-alternative splicing site within the third exon. Mutants deleted for UmRrm75 showed a slower growth rate than wild type strains in liquid and solid media, and their colonies showed a donut-like morphology on solid medium. Interestingly, although ΔUmRrm75 strains were not affected in filamentous growth induced by acid pH and oleic acid, they exhibited reduced mating, post-mating filamentous growth and virulence. Our data suggest that UmRrm75 is probably involved in cell growth, morphogenesis, and pathogenicity in U. maydis.

  3. Unh1, an Ustilago maydis Ndt80-like protein, controls completion of tumor maturation, teliospore development, and meiosis.

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    Doyle, Colleen E; Kitty Cheung, H Y; Spence, Kelsey L; Saville, Barry J

    2016-09-01

    In this study, Ustilago maydis Ndt80 homolog one, unh1, of the obligate sexual pathogen U. maydis,is described. Unh1 is the sole Ndt80-like DNA-binding protein inU. maydis. In this model basidiomycete, Unh1 plays a role in sexual development, influencing tumor maturation, teliospore development and subsequent meiotic completion. Teliospore formation was reduced in deletion mutants, and those that did form had unpigmented, hyaline cell walls, and germinated without completing meiosis. Constitutively expressing unh1 in haploid cells resulted in abnormal pigmentation, when grown in both potato dextrose broth and minimal medium, suggesting that pigmentation may be triggered by unh1 in U. maydis. The function of Unh1 in sexual development and pigment production depends on the presence of the Ndt80-like DNA-binding domain, identified within Unh1. In the absence of this domain, or when the binding domain was altered with targeted amino acid changes, ectopic expression of Unh1 failed to complement the unh1 deletion with regards to pigment production and sexual development. An investigation of U. maydis genes with upstream motifs similar to Ndt80 recognition sequences revealed that some have altered transcript levels in Δunh1 strains. We propose that the first characterized Ndt80-like DNA-binding protein in a basidiomycete, Unh1, acts as a transcription factor that is required for teliospore maturation and the completion of meiosis in U. maydis.

  4. Gene discovery and transcript analyses in the corn smut pathogen Ustilago maydis: expressed sequence tag and genome sequence comparison

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    Saville Barry J

    2007-09-01

    Full Text Available Abstract Background Ustilago maydis is the basidiomycete fungus responsible for common smut of corn and is a model organism for the study of fungal phytopathogenesis. To aid in the annotation of the genome sequence of this organism, several expressed sequence tag (EST libraries were generated from a variety of U. maydis cell types. In addition to utility in the context of gene identification and structure annotation, the ESTs were analyzed to identify differentially abundant transcripts and to detect evidence of alternative splicing and anti-sense transcription. Results Four cDNA libraries were constructed using RNA isolated from U. maydis diploid teliospores (U. maydis strains 518 × 521 and haploid cells of strain 521 grown under nutrient rich, carbon starved, and nitrogen starved conditions. Using the genome sequence as a scaffold, the 15,901 ESTs were assembled into 6,101 contiguous expressed sequences (contigs; among these, 5,482 corresponded to predicted genes in the MUMDB (MIPS Ustilago maydis database, while 619 aligned to regions of the genome not yet designated as genes in MUMDB. A comparison of EST abundance identified numerous genes that may be regulated in a cell type or starvation-specific manner. The transcriptional response to nitrogen starvation was assessed using RT-qPCR. The results of this suggest that there may be cross-talk between the nitrogen and carbon signalling pathways in U. maydis. Bioinformatic analysis identified numerous examples of alternative splicing and anti-sense transcription. While intron retention was the predominant form of alternative splicing in U. maydis, other varieties were also evident (e.g. exon skipping. Selected instances of both alternative splicing and anti-sense transcription were independently confirmed using RT-PCR. Conclusion Through this work: 1 substantial sequence information has been provided for U. maydis genome annotation; 2 new genes were identified through the discovery of 619

  5. The fungus Ustilago maydis and humans share disease-related proteins that are not found in Saccharomyces cerevisiae

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    Steinberg Gero

    2007-12-01

    Full Text Available Abstract Background The corn smut fungus Ustilago maydis is a well-established model system for molecular phytopathology. In addition, it recently became evident that U. maydis and humans share proteins and cellular processes that are not found in the standard fungal model Saccharomyces cerevisiae. This prompted us to do a comparative analysis of the predicted proteome of U. maydis, S. cerevisiae and humans. Results At a cut off at 20% identity over protein length, all three organisms share 1738 proteins, whereas both fungi share only 541 conserved proteins. Despite the evolutionary distance between U. maydis and humans, 777 proteins were shared. When applying a more stringent criterion (≥ 20% identity with a homologue in one organism over at least 50 amino acids and ≥ 10% less in the other organism, we found 681 proteins for the comparison of U. maydis and humans, whereas the both fungi share only 622 fungal specific proteins. Finally, we found that S. cerevisiae and humans shared 312 proteins. In the U. maydis to H. sapiens homology set 454 proteins are functionally classified and 42 proteins are related to serious human diseases. However, a large portion of 222 proteins are of unknown function. Conclusion The fungus U. maydis has a long history of being a model system for understanding DNA recombination and repair, as well as molecular plant pathology. The identification of functionally un-characterized genes that are conserved in humans and U. maydis opens the door for experimental work, which promises new insight in the cell biology of the mammalian cell.

  6. A chlorogenic acid esterase with a unique substrate specificity from Ustilago maydis.

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    Nieter, Annabel; Haase-Aschoff, Paul; Kelle, Sebastian; Linke, Diana; Krings, Ulrich; Popper, Lutz; Berger, Ralf G

    2015-03-01

    An extracellular chlorogenic acid esterase from Ustilago maydis (UmChlE) was purified to homogeneity by using three separation steps, including anion-exchange chromatography on a Q Sepharose FF column, preparative isoelectric focusing (IEF), and, finally, a combination of affinity chromatography and hydrophobic interaction chromatography on polyamide. SDS-PAGE analysis suggested a monomeric protein of ∼71 kDa. The purified enzyme showed maximal activity at pH 7.5 and at 37°C and was active over a wide pH range (3.5 to 9.5). Previously described chlorogenic acid esterases exhibited a comparable affinity for chlorogenic acid, but the enzyme from Ustilago was also active on typical feruloyl esterase substrates. Kinetic constants for chlorogenic acid, methyl p-coumarate, methyl caffeate, and methyl ferulate were as follows: Km values of 19.6 μM, 64.1 μM, 72.5 μM, and 101.8 μM, respectively, and kcat/Km values of 25.83 mM(-1) s(-1), 7.63 mM(-1) s(-1), 3.83 mM(-1) s(-1) and 3.75 mM(-1) s(-1), respectively. UmChlE released ferulic, p-coumaric, and caffeic acids from natural substrates such as destarched wheat bran (DSWB) and coffee pulp (CP), confirming activity on complex plant biomass. The full-length gene encoding UmChlE consisted of 1,758 bp, corresponding to a protein of 585 amino acids, and was functionally produced in Pichia pastoris GS115. Sequence alignments with annotated chlorogenic acid and feruloyl esterases underlined the uniqueness of this enzyme. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Ustilago maydis Rho1 and 14-3-3 homologues participate in pathways controlling cell separation and cell polarity.

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    Pham, Cau D; Yu, Zhanyang; Sandrock, Björn; Bölker, Michael; Gold, Scott E; Perlin, Michael H

    2009-07-01

    Proteins of the 14-3-3 and Rho-GTPase families are functionally conserved eukaryotic proteins that participate in many important cellular processes such as signal transduction, cell cycle regulation, malignant transformation, stress response, and apoptosis. However, the exact role(s) of these proteins in these processes is not entirely understood. Using the fungal maize pathogen, Ustilago maydis, we were able to demonstrate a functional connection between Pdc1 and Rho1, the U. maydis homologues of 14-3-3epsilon and Rho1, respectively. Our experiments suggest that Pdc1 regulates viability, cytokinesis, chromosome condensation, and vacuole formation. Similarly, U. maydis Rho1 is also involved in these three essential processes and exerts an additional function during mating and filamentation. Intriguingly, yeast two-hybrid and epistasis experiments suggest that both Pdc1 and Rho1 could be constituents of the same regulatory cascade(s) controlling cell growth and filamentation in U. maydis. Overexpression of rho1 ameliorated the defects of cells depleted for Pdc1. Furthermore, we found that another small G protein, Rac1, was a suppressor of lethality for both Pdc1 and Rho1. In addition, deletion of cla4, encoding a Rac1 effector kinase, could also rescue cells with Pdc1 depleted. Inferring from these data, we propose a model for Rho1 and Pdc1 functions in U. maydis.

  8. Pep1, a secreted effector protein of Ustilago maydis, is required for successful invasion of plant cells.

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    Gunther Doehlemann

    2009-02-01

    Full Text Available The basidiomycete Ustilago maydis causes smut disease in maize. Colonization of the host plant is initiated by direct penetration of cuticle and cell wall of maize epidermis cells. The invading hyphae are surrounded by the plant plasma membrane and proliferate within the plant tissue. We identified a novel secreted protein, termed Pep1, that is essential for penetration. Disruption mutants of pep1 are not affected in saprophytic growth and develop normal infection structures. However, Deltapep1 mutants arrest during penetration of the epidermal cell and elicit a strong plant defense response. Using Affymetrix maize arrays, we identified 116 plant genes which are differentially regulated in Deltapep1 compared to wild type infections. Most of these genes are related to plant defense. By in vivo immunolocalization, live-cell imaging and plasmolysis approaches, we detected Pep1 in the apoplastic space as well as its accumulation at sites of cell-to-cell passages. Site-directed mutagenesis identified two of the four cysteine residues in Pep1 as essential for function, suggesting that the formation of disulfide bridges is crucial for proper protein folding. The barley covered smut fungus Ustilago hordei contains an ortholog of pep1 which is needed for penetration of barley and which is able to complement the U. maydis Deltapep1 mutant. Based on these results, we conclude that Pep1 has a conserved function essential for establishing compatibility that is not restricted to the U. maydis / maize interaction.

  9. Characterization of a new aryl-alcohol oxidase secreted by the phytopathogenic fungus Ustilago maydis.

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    Couturier, Marie; Mathieu, Yann; Li, Ai; Navarro, David; Drula, Elodie; Haon, Mireille; Grisel, Sacha; Ludwig, Roland; Berrin, Jean-Guy

    2016-01-01

    The discovery of novel fungal lignocellulolytic enzymes is essential to improve the breakdown of plant biomass for the production of second-generation biofuels or biobased materials in green biorefineries. We previously reported that Ustilago maydis grown on maize secreted a diverse set of lignocellulose-acting enzymes including hemicellulases and putative oxidoreductases. One of the most abundant proteins of the secretome was a putative glucose-methanol-choline (GMC) oxidoreductase. The phylogenetic prediction of its function was hampered by the few characterized members within its clade. Therefore, we cloned the gene and produced the recombinant protein to high yield in Pichia pastoris. Functional screening using a library of substrates revealed that this enzyme was able to oxidize several aromatic alcohols. Of the tested aryl-alcohols, the highest oxidation rate was obtained with 4-anisyl alcohol. Oxygen, 1,4-benzoquinone, and 2,6-dichloroindophenol can serve as electron acceptors. This GMC oxidoreductase displays the characteristics of an aryl-alcohol oxidase (E.C.1.1.3.7), which is suggested to act on the lignin fraction in biomass.

  10. Appressorium formation in the corn smut fungus Ustilago maydis requires a G2 cell cycle arrest.

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    Castanheira, Sónia; Pérez-Martín, José

    2015-01-01

    Many of the most important plant diseases are caused by fungal pathogens that form specialized cell structures to breach the leaf surface as well as to proliferate inside the plant. To initiate pathogenic development, the fungus responds to a set of inductive cues. Some of them are of extracellular nature (environmental signals) while others respond to intracellular conditions (developmental signals). These signals have to be integrated into a single response that has as a major outcome changes in the morphogenesis of the fungus. The cell cycle regulation is pivotal during these cellular differentiations, and we hypothesized that cell cycle regulation would be likely to provide control points for infection development by fungal pathogens. Although efforts have been done in various fungal systems, there is still limited information available regarding the relationship of these processes with the induction of the virulence programs. Hence, the role of fungal cell cycle regulators -which are wide conserved elements- as true virulence factors, has yet to be defined. Here we discuss the recent finding that the formation of the appressorium, a structure required for plant penetration, in the corn smut fungus Ustilago maydis seems to be incompatible with an active cell cycle and, therefore genetic circuits evolved in this fungus to arrest the cell cycle during the growth of this fungus on plant surface, before the appressorium-mediated penetration into the plant tissue.

  11. Ustilago maydis natural antisense transcript expression alters mRNA stability and pathogenesis.

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    Donaldson, Michael E; Saville, Barry J

    2013-07-01

    Ustilago maydis infection of Zea mays leads to the production of thick-walled diploid teliospores that are the dispersal agent for this pathogen. Transcriptome analyses of this model biotrophic basidiomycete fungus identified natural antisense transcripts (NATs) complementary to 247 open reading frames. The U. maydis NAT cDNAs were fully sequenced and annotated. Strand-specific RT-PCR screens confirmed expression and identified NATs preferentially expressed in the teliospore. Targeted screens revealed four U. maydis NATs that are conserved in a related fungus. Expression of NATs in haploid cells, where they are not naturally occurring, resulted in increased steady-state levels of some complementary mRNAs. The expression of one NAT, as-um02151, in haploid cells resulted in a twofold increase in complementary mRNA levels, the formation of sense-antisense double-stranded RNAs, and unchanged Um02151 protein levels. This led to a model for NAT function in the maintenance and expression of stored teliospore mRNAs. In testing this model by deletion of the regulatory region, it was determined that alteration in NAT expression resulted in decreased pathogenesis in both cob and seedling infections. This annotation and functional analysis supports multiple roles for U. maydis NATs in controlling gene expression and influencing pathogenesis.

  12. Regulation of the expression of the whole genome of Ustilago maydis by a MAPK pathway.

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    Martínez-Soto, Domingo; Ruiz-Herrera, José

    2015-05-01

    The operation of mitogen-activated protein kinase (MAPK) signal transduction pathways is one of the most important mechanisms for the transfer of extracellular information into the cell. These pathways are highly conserved in eukaryotic organisms. In fungi, MAPK pathways are involved in the regulation of a number of cellular processes such as metabolism, homeostasis, pathogenesis and cell differentiation and morphogenesis. Considering the importance of pathways, in the present work we proceeded to identify all the genes that are regulated by the signal transduction pathway involved in mating, pathogenesis and morphogenesis of Ustilago maydis. Accordingly we made a comparison between the transcriptomes from a wild-type strain and an Ubc2 mutant affected in the interacting protein of this pathway by use of microarrays. By this methodology, we identified 939 genes regulated directly or indirectly by the MAPK pathway. Of them, 432 were positively, and 507 were negatively found regulated. By functional grouping, genes encoding cyclin-dependent kinases, transcription factors, proteins involved in signal transduction, in synthesis of wall and cell membrane, and involved in dimorphism were identified as differentially regulated. These data reveal the importance of these global studies, and the large (and unsuspected) number of functions of the fungus under the control of this MAPK, providing clues to the possible mechanisms involved.

  13. Production of itaconic acid by Ustilago maydis from agro wastes in solid state fermentation

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    MOKULA MD. RAFI

    2014-08-01

    Full Text Available Itaconic acid (IA is one of the hopeful substances within the cluster of organic acids. IA is used in artificial glass, bioactive compounds in pharmacy, medicine, agriculture, for the synthesis of fiber, resin, plastic, rubber, paints, surfactant, ion-exchange resins and lubricant. Most recurrently used microorganism for commercial production of IA is Aspergillus terreus. Some filamentous fungi belonging to Ustilaginales also produce IA. In the present work, an attempt was made to produce IA by Ustilago maydis employing Solid State Fermentation (SSF from various agro wastes like ground nut shells, rice bran, rice husk, orange pulp, ground nut oil cake, orange pulp and sugarcane bagasse as carbon substrates, which were used after pretreatment. 10 g of each substrate was taken in a 500 ml conical flasks separately and supplemented with 20 mL nutrient solution containing glucose, at pH 3. One milliliter inoculum containing 1×107 spores was added and moisture was maintained at 60%. After incubation at 32°C for 5 days, the acid production was estimated by spectrophotometric method and by HPLC analysis. Interestingly, the yield of itaconic acid was promising with all the above substrates, where orange pulp, sugarcane bagasse and rice bran supported higher yields.

  14. Differential chromosomal and mitochondrial DNA synthesis in temperature-sensitive mutants of Ustilago maydis

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    Unrau, P.

    1977-01-01

    The amount and type of residual DNA synthesis was determined in eight temperature-sensitive mutants of the smut fungus Ustilago maydis after incubation at the restrictive temperature (32/sup 0/C) for eight hours. Mutants ts-220, ts-207, ts-432 and ts-346 were found to have an overall reduction in the synthesis of both nuclear and mitochondrial DNA in comparison to the wild-type. In mutants ts-20, tsd 1-1, ts-84 and pol 1-1 nuclear DNA synthesis was depressed relative to mitochondrial synthesis. The DNA-polymerase mutant pol 1-1 had persistent nuclear synthesis at about 50% of the rate of synthesis of mitochondrial DNA and similar behavior was observed in a diploid homozygous strain. Mutant ts-84 had an initial burst of DNA synthesis which was reduced for nuclear but not mitochondrial synthesis after three hours preincubation at 32/sup 0/C. tsd 1-1 and ts-20 had nuclear residual synthesis amounting to about 25% of the relative rate of mitochondrial synthesis which correlates to increasing UV sensitivity of these strains on incubation at 32/sup 0/C. A pol 1-1 ts-84 double mutant had an additive loss of nuclear DNA synthesis which indicates that the steps of replication involved may be sequential.

  15. Regulation of genes involved in cell wall synthesis and structure during Ustilago maydis dimorphism.

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    Robledo-Briones, Mariana; Ruiz-Herrera, José

    2013-02-01

    The cell wall is the structure that provides the shape to fungal cells and protects them from the difference in osmotic pressure existing between the cytosol and the external medium. Accordingly, changes in structure and composition of the fungal wall must occur during cell differentiation, including the dimorphic transition of fungi. We analyzed, by use of microarrays, the transcriptional regulation of the 639 genes identified to be involved in cell wall synthesis and structure plus the secretome of the Basidiomycota species Ustilago maydis during its dimorphic transition induced by a change in pH. Of these, 189 were differentially expressed during the process, and using as control two monomorphic mutants, one yeast like and the other mycelium constitutive, 66 genes specific of dimorphism were identified. Most of these genes were up-regulated in the mycelial phase. These included CHS genes, genes involved in β-1,6-glucan synthesis, N-glycosylation, and proteins containing a residue of glycosylphosphatidylinositol, and a number of genes from the secretome. The possible significance of these data on cell wall plasticity is discussed.

  16. Applying unconventional secretion of the endochitinase Cts1 to export heterologous proteins in Ustilago maydis.

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    Stock, Janpeter; Sarkari, Parveen; Kreibich, Saskia; Brefort, Thomas; Feldbrügge, Michael; Schipper, Kerstin

    2012-10-15

    The demand on the biotechnological production of proteins for pharmaceutical, medical and industrial applications is steadily growing. For the production of challenging proteins, we aim to establish a novel expression platform in the well characterized eukaryotic microorganism Ustilago maydis. In filaments of this fungus, secretion of the endochitinase Cts1 depends on mRNA transport along microtubules, which is mediated by the key RNA-binding protein Rrm4. Here, we report two important findings: (i) Cts1 secretion occurs via a novel unconventional route and (ii) this secretory mechanism can be exploited for the export of active heterologous proteins. Initially, we used β-glucuronidase (Gus) as a reporter for unconventional secretion. This bacterial enzyme is inactivated by N-glycosylation during its passage through the conventional eukaryotic secretory pathway. By contrast, in our system Gus was exported in its active form by fusion to Cts1 confirming its secretion by an unconventional route. As a proof-of-principle for economically important biopharmaceuticals we expressed an active single-chain antibody. Importantly, the novel protein export pathway circumvents N-glycosylation which is advantageous in many applications, e.g., to avoid undesired immune reactions in humans. Thus, the unconventional Cts1 secretion machinery has a high potential for the production of biotechnologically relevant proteins.

  17. Applying Unconventional Secretion in Ustilago maydis for the Export of Functional Nanobodies

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    Marius Terfrüchte

    2017-04-01

    Full Text Available Exploiting secretory pathways for production of heterologous proteins is highly advantageous with respect to efficient downstream processing. In eukaryotic systems the vast majority of heterologous proteins for biotechnological application is exported via the canonical endoplasmic reticulum–Golgi pathway. In the endomembrane system target proteins are often glycosylated and may thus be modified with foreign glycan patterns. This can be destructive for their activity or cause immune reactions against therapeutic proteins. Hence, using unconventional secretion for protein expression is an attractive alternative. In the fungal model Ustilago maydis, chitinase Cts1 is secreted via an unconventional pathway connected to cell separation which can be used to co-export heterologous proteins. Here, we apply this mechanism for the production of nanobodies. First, we achieved expression and unconventional secretion of a functional nanobody directed against green fluorescent protein (Gfp. Second, we found that Cts1 binds to chitin and that this feature can be applied to generate a Gfp-trap. Thus, we demonstrated the dual use of Cts1 serving both as export vehicle and as purification tag. Finally, we established and optimized the production of a nanobody against botulinum toxin A and hence describe the first pharmaceutically relevant target exported by Cts1-mediated unconventional secretion.

  18. An unusual MAP kinase is required for efficient penetration of the plant surface by Ustilago maydis.

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    Brachmann, Andreas; Schirawski, Jan; Müller, Philip; Kahmann, Regine

    2003-05-01

    In Ustilago maydis, pathogenic development is controlled by a heterodimer of the two homeodomain proteins bW and bE. We have identified by RNA fingerprinting a b-regulated gene, kpp6, which encodes an unusual MAP kinase. Kpp6 is similar to a number of other fungal MAP kinases involved in mating and pathogenicity, but contains an additional N-terminal domain unrelated to other proteins. Transcription of the kpp6 gene yields two transcripts differing in length, but encoding proteins of identical mass. One transcript is upregulated by the bW/bE heterodimer, while the other is induced after pheromone stimulation. kpp6 deletion mutants are attenuated in pathogenicity. kpp6(T355A,Y357F) mutants carrying a non-activatable allele of kpp6 are more severely compromised in pathogenesis. These strains can still form appressoria, but are defective in the subsequent penetration of the plant cuticle. Kpp6 is expressed during all stages of the sexual life cycle except mature spores. We speculate that Kpp6 may respond to a plant signal and regulate the genes necessary for efficient penetration of plant tissue.

  19. Nucleic acid-binding glycoproteins which solubilize nucleic acids in dilute acid: re-examination of the Ustilago maydis glycoproteins

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    Unrau, P.; Champ, D.R.; Young, J.L.; Grant, C.E.

    1980-01-01

    Holloman reported the isolation from Ustilago maydis of a glycoprotein which prevented the precipitation of nucleic acids in cold 5% trichloroacetic acid. Two glycoprotein fractions from U. maydis with this nucleic acid-solubilizing activity were isolated in our laboratory using improved purification procedures. The activity was not due to nuclease contamination. The glycoproteins are distinguished by: their ability to bind to concanavalin A-Sepharose; their differential binding to double- and single-stranded deoxyribonucleic acid, and to ribonucleic acid; their molecular weights (46,000 and 69,000); and the relative amounts present in growing versus nongrowing cells. Both fractions required sulfhydryl-reducing conditions for optimal yields, specific activity, and stability. Nucleic acid binding was cooperative, the minimum number of glycoproteins required to make a native T7 DNA molecule soluble in dilute acid being estimated at 2 and 15, respectively.

  20. Characterization of ApB73, a virulence factor important for colonization of Zea mays by the smut Ustilago maydis.

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    Stirnberg, Alexandra; Djamei, Armin

    2016-12-01

    The biotrophic fungus Ustilago maydis, the causal agent of corn smut disease, uses numerous small secreted effector proteins to suppress plant defence responses and reshape the host metabolism. However, the role of specific effectors remains poorly understood. Here, we describe the identification of ApB73 (Apathogenic in B73), an as yet uncharacterized protein essential for the successful colonization of maize by U. maydis. We show that apB73 is transcriptionally induced during the biotrophic stages of the fungal life cycle. The deletion of the apB73 gene results in cultivar-specific loss of gall formation in the host. The ApB73 protein is conserved among closely related smut fungi. However, using virulence assays, we show that only the orthologue of the maize-infecting head smut Sporisorium reilianum can complement the mutant phenotype of U. maydis. Although microscopy shows that ApB73 is secreted into the biotrophic interface, it seems to remain associated with fungal cell wall components or the fungal plasma membrane. Taken together, the results show that ApB73 is a conserved and important virulence factor of U. maydis that localizes to the interface between the pathogen and its host Zea mays.

  1. Compatibility in the Ustilago maydis-maize interaction requires inhibition of host cysteine proteases by the fungal effector Pit2.

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    André N Mueller

    2013-02-01

    Full Text Available The basidiomycete Ustilago maydis causes smut disease in maize, with large plant tumors being formed as the most prominent disease symptoms. During all steps of infection, U. maydis depends on a biotrophic interaction, which requires an efficient suppression of plant immunity. In a previous study, we identified the secreted effector protein Pit2, which is essential for maintenance of biotrophy and induction of tumors. Deletion mutants for pit2 successfully penetrate host cells but elicit various defense responses, which stops further fungal proliferation. We now show that Pit2 functions as an inhibitor of a set of apoplastic maize cysteine proteases, whose activity is directly linked with salicylic-acid-associated plant defenses. Consequently, protease inhibition by Pit2 is required for U. maydis virulence. Sequence comparisons with Pit2 orthologs from related smut fungi identified a conserved sequence motif. Mutation of this sequence caused loss of Pit2 function. Consequently, expression of the mutated protein in U. maydis could not restore virulence of the pit2 deletion mutant, indicating that the protease inhibition by Pit2 is essential for fungal virulence. Moreover, synthetic peptides of the conserved sequence motif showed full activity as protease inhibitor, which identifies this domain as a new, minimal protease inhibitor domain in plant-pathogenic fungi.

  2. Compatibility in the Ustilago maydis-maize interaction requires inhibition of host cysteine proteases by the fungal effector Pit2.

    Science.gov (United States)

    Mueller, André N; Ziemann, Sebastian; Treitschke, Steffi; Aßmann, Daniela; Doehlemann, Gunther

    2013-02-01

    The basidiomycete Ustilago maydis causes smut disease in maize, with large plant tumors being formed as the most prominent disease symptoms. During all steps of infection, U. maydis depends on a biotrophic interaction, which requires an efficient suppression of plant immunity. In a previous study, we identified the secreted effector protein Pit2, which is essential for maintenance of biotrophy and induction of tumors. Deletion mutants for pit2 successfully penetrate host cells but elicit various defense responses, which stops further fungal proliferation. We now show that Pit2 functions as an inhibitor of a set of apoplastic maize cysteine proteases, whose activity is directly linked with salicylic-acid-associated plant defenses. Consequently, protease inhibition by Pit2 is required for U. maydis virulence. Sequence comparisons with Pit2 orthologs from related smut fungi identified a conserved sequence motif. Mutation of this sequence caused loss of Pit2 function. Consequently, expression of the mutated protein in U. maydis could not restore virulence of the pit2 deletion mutant, indicating that the protease inhibition by Pit2 is essential for fungal virulence. Moreover, synthetic peptides of the conserved sequence motif showed full activity as protease inhibitor, which identifies this domain as a new, minimal protease inhibitor domain in plant-pathogenic fungi.

  3. Identificación de las proteínas secretadas por el hongo Ustilago maydis (DeCandole Corda (Basidiomiceto cultivado en condiciones in vitro

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    Andrés Adolfo Estrada-Luna

    2010-01-01

    Full Text Available Introducción: Ustilago maydis es un hongo basidiomiceto que infecta al maíz y teozintle produciendo una enfermedad conocida como carbón común o huitlacoche. Actualmente no existen reportes acerca del secretoma del hongo cultivado bajo condiciones in vitro. Un estudio de esta naturaleza permitiría caracterizar los genes involucrados en varios procesos importantes, entre los que se tienen aquellos relacionados con la nutrición, la patogenicidad y la diferenciación del hongo. El objetivo de esta investigación fue identificar las proteínas secretadas al medio de cultivo por las formas de levadura o micelio de este hongo cultivado en dos condiciones de pH. Método: Se generaron las formas de micelio o levadura de Ustilago maydis (cepa FB2¿a2b2 a través del cultivo en medios mínimos con pH 3 y 7 respectivamente y se determinó su cinética de crecimiento. Las proteínas secretadas al medio se concentraron en una columna de fase reversa Sep-Pak Plus C18 y se eluyeron con una solución de acetonitrilo (60 % + ácido trifluoroacético (0.1 %, seguida de su liofilización parcial, y precipitación con ácido tricloroacético-acetona. Posteriormente las muestras fueron sometidas a electroforesis en poliacrilamida (SDS-PAGE y los geles teñidos con azul de Coomassie. Las bandas de proteína se cortaron del gel y se digirieron con tripsina. Las mezclas de péptidos fueron inyectados para su análisis en un espectrómetro de masas y el espectro MS/MS obtenido fue procesado en Masslynx 4.0 antes de someterlo al programa MASCOT (Matrix Science para realizar las búsquedas no-redundantes en la base de datos del National Center for Biotechnology Information. Resultados: El crecimiento de U. maydis a pH 7 fue mayor que a pH 3 (D.O. a 600 nm= 1.35 y 0.85, respectivamente a las 30 h de incubación. El proceso dimórfico de levadura a micelio a pH 3 se inició a las 8 h después de iniciados los cultivos. A las 30 h de cultivo se observó que el 100 % de

  4. The impact of common smut(Ustilago maydis) on aflatoxin and fumonisin in transgenic Bt and non-Bt maize (Zea mays)

    Science.gov (United States)

    Corn infected with Ustilago maydis (common smut), produces galls that are valued food in certain cultures, but may be contaminated with mycotoxins. Field studies conducted in Elizabeth, Mississippi used near-isogenic Bt and non-Bt corn hybrids. The levels of aflatoxin and fumonisin were determined ...

  5. The Filament-specific Rep1-1 Repellent of the Phytopathogen Ustilago maydis Forms Functional Surface-active Amyloid-like Fibrils

    NARCIS (Netherlands)

    Teertstra, Wieke R.; van der Velden, Gisela J.; de Jong, Jan F.; Kruijtzer, John A. W.; Liskamp, Rob M. J.; Kroon-Batenburg, Loes M. J.; Muller, Wally H.; Gebbink, Martijn F. B. G.; Wosten, Han A. B.

    2009-01-01

    Repellents of the maize pathogen Ustilago maydis are involved in formation of hydrophobic aerial hyphae and in cellular attachment. These peptides, called Rep1-1 to Rep1-11, are encoded by the rep1 gene and result from cleavage of the precursor protein Rep1 during passage of the secretion pathway. U

  6. The Filament-specific Rep1-1 Repellent of the Phytopathogen Ustilago maydis Forms Functional Surface-active Amyloid-like Fibrils

    NARCIS (Netherlands)

    Teertstra, Wieke R.; van der Velden, Gisela J.; de Jong, Jan F.; Kruijtzer, John A. W.; Liskamp, Rob M. J.; Kroon-Batenburg, Loes M. J.; Muller, Wally H.; Gebbink, Martijn F. B. G.; Wosten, Han A. B.

    2009-01-01

    Repellents of the maize pathogen Ustilago maydis are involved in formation of hydrophobic aerial hyphae and in cellular attachment. These peptides, called Rep1-1 to Rep1-11, are encoded by the rep1 gene and result from cleavage of the precursor protein Rep1 during passage of the secretion pathway. U

  7. The cell end marker Tea4 regulates morphogenesis and pathogenicity in the basidiomycete fungus Ustilago maydis.

    Science.gov (United States)

    Valinluck, Michael; Woraratanadharm, Tad; Lu, Ching-yu; Quintanilla, Rene H; Banuett, Flora

    2014-05-01

    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.

  8. The basidiomycete Ustilago maydis has two plasma membrane H⁺-ATPases related to fungi and plants.

    Science.gov (United States)

    Robles-Martínez, Leobarda; Pardo, Juan Pablo; Miranda, Manuel; Mendez, Tavis L; Matus-Ortega, Macario Genaro; Mendoza-Hernández, Guillermo; Guerra-Sánchez, Guadalupe

    2013-10-01

    The fungal and plant plasma membrane H⁺-ATPases play critical roles in the physiology of yeast, plant and protozoa cells. We identified two genes encoding two plasma membrane H⁺-ATPases in the basidiomycete Ustilago maydis, one protein with higher identity to fungal (um02581) and the other to plant (um01205) H⁺-ATPases. Proton pumping activity was 5-fold higher when cells were grown in minimal medium with ethanol compared to cells cultured in rich YPD medium, but total vanadate-sensitive ATPase activity was the same in both conditions. In contrast, the activity in cells cultured in minimal medium with glucose was 2-fold higher than in YPD or ethanol, implicating mechanisms for the regulation of the plasma membrane ATPase activity in U. maydis. Analysis of gene expression of the H⁺-ATPases from cells grown under different conditions, showed that the transcript expression of um01205 (plant-type) was higher than that of um02581 (fungal-type). The translation of the two proteins was confirmed by mass spectrometry analysis. Unlike baker's yeast and plant H⁺-ATPases, where the activity is increased by a short incubation with glucose or sucrose, respectively, U. maydis H⁺-ATPase activity did not change in response to these sugars. Sequence analysis of the two U. maydis H⁺-ATPases revealed the lack of canonical threonine and serine residues which are targets of protein kinases in Saccharomyces cerevisiae and Arabidopsis thaliana plasma membrane H⁺-ATPases, suggesting that phosphorylation of the U. maydis enzymes occurs at different amino acid residues.

  9. Tartronate semialdehyde reductase defines a novel rate-limiting step in assimilation and bioconversion of glycerol in Ustilago maydis.

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    Yanbin Liu

    Full Text Available BACKGROUND: Glycerol is a by-product of biodiesel production. Currently, it has limited applications with low bioconversion efficiency to most metabolites reported. This is partly attributed to the poor knowledge on the glycerol metabolic pathway in bacteria and fungi. METHODOLOGY/PRINCIPAL FINDINGS: We have established a fast screening method for identification of genes that improve glycerol utilization in Ustilago maydis. This was done by comparing the growth rates of T-DNA tagged mutant colonies on solid medium using glycerol as the sole carbon source. We present a detailed characterization of one of the mutants, GUM1, which contains a T-DNA element inserted into the promoter region of UM02592 locus (MIPS Ustilago maydis database, MUMDB, leading to enhanced and constitutive expression of its mRNA. We have demonstrated that um02592 encodes a functional tartronate semialdehyde reductase (Tsr1, which showed dual specificity to cofactors NAD(+ and NADP(+ and strong substrate specificity and enantioselectivity for D-glycerate. Improved glycerol assimilation in GUM1 was associated with elevated expression of tsr1 mRNA and this could be phenocopied by over-expression of the gene. Glycolipid accumulation was reduced by 45.2% in the knockout mutant whereas introduction of an extra copy of tsr1 driven by the glyceraldehyde phosphate dehydrogenase promoter increased it by 40.4%. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that tartronate semialdehyde reductase (TSR plays an important role in glycerol assimilation in U. maydis and defines a novel target in genetic engineering for improved conversion of glycerol to higher value products. Our results add significant depth to the understanding of the glycerol metabolic pathway in fungi. We have demonstrated, for the first time, a biological role of a eukaryotic TSR.

  10. Uml2 is a novel CalB-type lipase of Ustilago maydis with phospholipase A activity.

    Science.gov (United States)

    Buerth, Christoph; Kovacic, Filip; Stock, Janpeter; Terfrüchte, Marius; Wilhelm, Susanne; Jaeger, Karl-Erich; Feldbrügge, Michael; Schipper, Kerstin; Ernst, Joachim F; Tielker, Denis

    2014-06-01

    CalB of Pseudozyma aphidis (formerly named Candida antarctica) is one of the most widely applied enzymes in industrial biocatalysis. Here, we describe a protein with 66 % sequence identity to CalB, designated Ustilago maydis lipase 2 (Uml2), which was identified as the product of gene um01422 of the corn smut fungus U. maydis. Sequence analysis of Uml2 revealed the presence of a typical lipase catalytic triad, Ser-His-Asp with Ser125 located in a Thr-Xaa-Ser-Xaa-Gly pentapeptide. Deletion of the uml2 gene in U. maydis diminished the ability of cells to hydrolyse fatty acids from tributyrin or Tween 20/80 substrates, thus demonstrating that Uml2 functions as a lipase that may contribute to nutrition of this fungal pathogen. Uml2 was heterologously produced in Pichia pastoris and recombinant N-glycosylated Uml2 protein was purified from the culture medium. Purified Uml2 released short- and long-chain fatty acids from p-nitrophenyl esters and Tween 20/80 substrates. Furthermore, phosphatidylcholine substrates containing long-chain saturated or unsaturated fatty acids were effectively hydrolysed. Both esterase and phospholipase A activity of Uml2 depended on the Ser125 catalytic residue. These results indicate that Uml2, in contrast to CalB, exhibits not only esterase and lipase activity but also phospholipase A activity. Thus, by genome mining, we identified a novel CalB-like lipase with different substrate specificities.

  11. The Hos2 Histone Deacetylase Controls Ustilago maydis Virulence through Direct Regulation of Mating-Type Genes.

    Science.gov (United States)

    Elías-Villalobos, Alberto; Fernández-Álvarez, Alfonso; Moreno-Sánchez, Ismael; Helmlinger, Dominique; Ibeas, José I

    2015-08-01

    Morphological changes are critical for host colonisation in plant pathogenic fungi. These changes occur at specific stages of their pathogenic cycle in response to environmental signals and are mediated by transcription factors, which act as master regulators. Histone deacetylases (HDACs) play crucial roles in regulating gene expression, for example by locally modulating the accessibility of chromatin to transcriptional regulators. It has been reported that HDACs play important roles in the virulence of plant fungi. However, the specific environment-sensing pathways that control fungal virulence via HDACs remain poorly characterised. Here we address this question using the maize pathogen Ustilago maydis. We find that the HDAC Hos2 is required for the dimorphic switch and pathogenic development in U. maydis. The deletion of hos2 abolishes the cAMP-dependent expression of mating type genes. Moreover, ChIP experiments detect Hos2 binding to the gene bodies of mating-type genes, which increases in proportion to their expression level following cAMP addition. These observations suggest that Hos2 acts as a downstream component of the cAMP-PKA pathway to control the expression of mating-type genes. Interestingly, we found that Clr3, another HDAC present in U. maydis, also contributes to the cAMP-dependent regulation of mating-type gene expression, demonstrating that Hos2 is not the only HDAC involved in this control system. Overall, our results provide new insights into the role of HDACs in fungal phytopathogenesis.

  12. The Hos2 Histone Deacetylase Controls Ustilago maydis Virulence through Direct Regulation of Mating-Type Genes.

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    Alberto Elías-Villalobos

    2015-08-01

    Full Text Available Morphological changes are critical for host colonisation in plant pathogenic fungi. These changes occur at specific stages of their pathogenic cycle in response to environmental signals and are mediated by transcription factors, which act as master regulators. Histone deacetylases (HDACs play crucial roles in regulating gene expression, for example by locally modulating the accessibility of chromatin to transcriptional regulators. It has been reported that HDACs play important roles in the virulence of plant fungi. However, the specific environment-sensing pathways that control fungal virulence via HDACs remain poorly characterised. Here we address this question using the maize pathogen Ustilago maydis. We find that the HDAC Hos2 is required for the dimorphic switch and pathogenic development in U. maydis. The deletion of hos2 abolishes the cAMP-dependent expression of mating type genes. Moreover, ChIP experiments detect Hos2 binding to the gene bodies of mating-type genes, which increases in proportion to their expression level following cAMP addition. These observations suggest that Hos2 acts as a downstream component of the cAMP-PKA pathway to control the expression of mating-type genes. Interestingly, we found that Clr3, another HDAC present in U. maydis, also contributes to the cAMP-dependent regulation of mating-type gene expression, demonstrating that Hos2 is not the only HDAC involved in this control system. Overall, our results provide new insights into the role of HDACs in fungal phytopathogenesis.

  13. Interactions between Fusarium verticillioides, Ustilago maydis, and Zea mays: an endophyte, a pathogen, and their shared plant host.

    Science.gov (United States)

    Rodriguez Estrada, Alma E; Jonkers, Wilfried; Kistler, H Corby; May, Georgiana

    2012-07-01

    Highly diverse communities of microbial symbionts occupy eukaryotic organisms, including plants. While many well-studied symbionts may be characterized as either parasites or as mutualists, the prevalent but cryptic endophytic fungi are less easily qualified because they do not cause observable symptoms of their presence within their host. Here, we investigate the interactions of an endophytic fungus, Fusarium verticillioides with a pathogen, Ustilago maydis, as they occur within maize (Zea mays). We used experimental inoculations to evaluate metabolic mechanisms by which these three organisms might interact. We assessed the impacts of fungal-fungal interactions on endophyte and pathogen growth within the plant, and on plant growth. We find that F. verticillioides modulates the growth of U. maydis and thus decreases the pathogen's aggressiveness toward the plant. With co-inoculation of the endophyte with the pathogen, plant growth is similar to that which would be gained without the pathogen present. However, the endophyte may also break down plant compounds that limit U. maydis growth, and obtains a growth benefit from the presence of the pathogen. Thus, an endophyte such as F. verticillioides may function as both a defensive mutualist and a parasite, and express nutritional modes that depend on ecological context. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Activation of the cell wall integrity pathway promotes escape from G2 in the fungus Ustilago maydis.

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    Natalia Carbó

    2010-07-01

    Full Text Available It is widely accepted that MAPK activation in budding and fission yeasts is often associated with negative effects on cell cycle progression, resulting in delay or arrest at a specific stage in the cell cycle, thereby enabling cells to adapt to changing environmental conditions. For instance, activation of the Cell Wall Integrity (CWI pathway in the budding yeast Saccharomyces cerevisiae signals an increase in CDK inhibitory phosphorylation, which leads cells to remain in the G2 phase. Here we characterized the CWI pathway of Ustilago maydis, a fungus evolutionarily distant from budding and fission yeasts, and show that activation of the CWI pathway forces cells to escape from G2 phase. In spite of these disparate cell cycle responses in S. cerevisiae and U. maydis, the CWI pathway in both organisms appears to respond to the same class cell wall stressors. To understand the basis of such a difference, we studied the mechanism behind the U. maydis response. We found that activation of CWI pathway in U. maydis results in a decrease in CDK inhibitory phosphorylation, which depends on the mitotic phosphatase Cdc25. Moreover, in response to activation of the CWI pathway, Cdc25 accumulates in the nucleus, providing a likely explanation for the increase in the unphosphorylated form of CDK. We also found that the extended N-terminal domain of Cdc25, which is dispensable under normal growth conditions, is required for this G2 escape as well as for resistance to cell wall stressors. We propose that the process of cell cycle adaptation to cell stress evolved differently in these two divergent organisms so that each can move towards a cell cycle phase most appropriate for responding to the environmental signals encountered.

  15. Ustilago maydis killer toxin as a new tool for the biocontrol of the wine spoilage yeast Brettanomyces bruxellensis.

    Science.gov (United States)

    Santos, Antonio; Navascués, Eva; Bravo, Enrique; Marquina, Domingo

    2011-01-31

    Brettanomyces bruxellensis is one of the most damaging species for wine quality, and tools for controlling its growth are limited. In this study, thirty-nine strains belonging to Saccharomyces cerevisiae and B. bruxellensis have been isolated from wineries, identified and then tested against a panel of thirty-nine killer yeasts. Here, for the first time, the killer activity of Ustilago maydis is proven to be effective against B. bruxellensis. Mixed cultures in winemaking conditions show that U. maydis CYC 1410 has the ability to inhibit B. bruxellensis, while S. cerevisiae is fully resistant to its killer activity, indicating that it could be used in wine fermentation to avoid the development of B. bruxellensis without undesirable effects on the fermentative yeast. The characterization of the dsRNAs isolated and purified from U. maydis CYC 1410 indicated that this strain produces a KP6-related toxin. Killer toxin extracts were active against B. bruxellensis at pH values between 3.0 and 4.5 and temperatures comprised between 15 °C and 25 °C, confirming their biocontrol activity in winemaking and wine aging conditions. Furthermore, small amounts (100 AU/ml) of killer toxin extracts from U. maydis significantly reduced the amount of 4-ethylphenol produced by B. bruxellensis, indicating that in addition to the growth inhibition observed for high killer toxin concentrations (ranging from 400 to 2000 AU/ml), small amounts of the toxin are able to reduce the production of volatile phenols responsible for the aroma defects in wines caused by B. bruxellensis.

  16. Activation of the cell wall integrity pathway promotes escape from G2 in the fungus Ustilago maydis.

    Science.gov (United States)

    Carbó, Natalia; Pérez-Martín, José

    2010-07-01

    It is widely accepted that MAPK activation in budding and fission yeasts is often associated with negative effects on cell cycle progression, resulting in delay or arrest at a specific stage in the cell cycle, thereby enabling cells to adapt to changing environmental conditions. For instance, activation of the Cell Wall Integrity (CWI) pathway in the budding yeast Saccharomyces cerevisiae signals an increase in CDK inhibitory phosphorylation, which leads cells to remain in the G2 phase. Here we characterized the CWI pathway of Ustilago maydis, a fungus evolutionarily distant from budding and fission yeasts, and show that activation of the CWI pathway forces cells to escape from G2 phase. In spite of these disparate cell cycle responses in S. cerevisiae and U. maydis, the CWI pathway in both organisms appears to respond to the same class cell wall stressors. To understand the basis of such a difference, we studied the mechanism behind the U. maydis response. We found that activation of CWI pathway in U. maydis results in a decrease in CDK inhibitory phosphorylation, which depends on the mitotic phosphatase Cdc25. Moreover, in response to activation of the CWI pathway, Cdc25 accumulates in the nucleus, providing a likely explanation for the increase in the unphosphorylated form of CDK. We also found that the extended N-terminal domain of Cdc25, which is dispensable under normal growth conditions, is required for this G2 escape as well as for resistance to cell wall stressors. We propose that the process of cell cycle adaptation to cell stress evolved differently in these two divergent organisms so that each can move towards a cell cycle phase most appropriate for responding to the environmental signals encountered.

  17. The snf1 gene of Ustilago maydis acts as a dual regulator of cell wall degrading enzymes.

    Science.gov (United States)

    Nadal, Marina; Garcia-Pedrajas, Maria D; Gold, Scott E

    2010-12-01

    Many fungal plant pathogens are known to produce extracellular enzymes that degrade cell wall elements required for host penetration and infection. Due to gene redundancy, single gene deletions generally do not address the importance of these enzymes in pathogenicity. Cell wall degrading enzymes (CWDEs) in fungi are often subject to carbon catabolite repression at the transcriptional level such that, when glucose is available, CWDE-encoding genes, along with many other genes, are repressed. In Saccharomyces cerevisiae, one of the main players controlling this process is SNF1, which encodes a protein kinase. In this yeast, Snf1p is required to release glucose repression when this sugar is depleted from the growth medium. We have employed a reverse genetic approach to explore the role of the SNF1 ortholog as a potential regulator of CWDE gene expression in Ustilago maydis. We identified U. maydis snf1 and deleted it from the fungal genome. Consistent with our hypothesis, the relative expression of an endoglucanase and a pectinase was higher in the wild type than in the Δsnf1 mutant strain when glucose was depleted from the growth medium. However, when cells were grown in derepressive conditions, the relative expression of two xylanase genes was unexpectedly higher in the Δsnf1 strain than in the wild type, indicating that, in this case, snf1 negatively regulated the expression of these genes. Additionally, we found that, contrary to several other fungal species, U. maydis Snf1 was not required for utilization of alternative carbon sources. Also, unlike in ascomycete plant pathogens, deletion of snf1 did not profoundly affect virulence in U. maydis.

  18. In Silico Analysis of the Structural and Biochemical Features of the NMD Factor UPF1 in Ustilago maydis.

    Science.gov (United States)

    Martínez-Montiel, Nancy; Morales-Lara, Laura; Hernández-Pérez, Julio M; Martínez-Contreras, Rebeca D

    2016-01-01

    The molecular mechanisms regulating the accuracy of gene expression are still not fully understood. Among these mechanisms, Nonsense-mediated Decay (NMD) is a quality control process that detects post-transcriptionally abnormal transcripts and leads them to degradation. The UPF1 protein lays at the heart of NMD as shown by several structural and functional features reported for this factor mainly for Homo sapiens and Saccharomyces cerevisiae. This process is highly conserved in eukaryotes but functional diversity can be observed in various species. Ustilago maydis is a basidiomycete and the best-known smut, which has become a model to study molecular and cellular eukaryotic mechanisms. In this study, we performed in silico analysis to investigate the structural and biochemical properties of the putative UPF1 homolog in Ustilago maydis. The putative homolog for UPF1 was recognized in the annotated genome for the basidiomycete, exhibiting 66% identity with its human counterpart at the protein level. The known structural and functional domains characteristic of UPF1 homologs were also found. Based on the crystal structures available for UPF1, we constructed different three-dimensional models for umUPF1 in order to analyze the secondary and tertiary structural features of this factor. Using these models, we studied the spatial arrangement of umUPF1 and its capability to interact with UPF2. Moreover, we identified the critical amino acids that mediate the interaction of umUPF1 with UPF2, ATP, RNA and with UPF1 itself. Mutating these amino acids in silico showed an important effect over the native structure. Finally, we performed molecular dynamic simulations for UPF1 proteins from H. sapiens and U. maydis and the results obtained show a similar behavior and physicochemical properties for the protein in both organisms. Overall, our results indicate that the putative UPF1 identified in U. maydis shows a very similar sequence, structural organization, mechanical stability

  19. In Silico Analysis of the Structural and Biochemical Features of the NMD Factor UPF1 in Ustilago maydis.

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    Nancy Martínez-Montiel

    Full Text Available The molecular mechanisms regulating the accuracy of gene expression are still not fully understood. Among these mechanisms, Nonsense-mediated Decay (NMD is a quality control process that detects post-transcriptionally abnormal transcripts and leads them to degradation. The UPF1 protein lays at the heart of NMD as shown by several structural and functional features reported for this factor mainly for Homo sapiens and Saccharomyces cerevisiae. This process is highly conserved in eukaryotes but functional diversity can be observed in various species. Ustilago maydis is a basidiomycete and the best-known smut, which has become a model to study molecular and cellular eukaryotic mechanisms. In this study, we performed in silico analysis to investigate the structural and biochemical properties of the putative UPF1 homolog in Ustilago maydis. The putative homolog for UPF1 was recognized in the annotated genome for the basidiomycete, exhibiting 66% identity with its human counterpart at the protein level. The known structural and functional domains characteristic of UPF1 homologs were also found. Based on the crystal structures available for UPF1, we constructed different three-dimensional models for umUPF1 in order to analyze the secondary and tertiary structural features of this factor. Using these models, we studied the spatial arrangement of umUPF1 and its capability to interact with UPF2. Moreover, we identified the critical amino acids that mediate the interaction of umUPF1 with UPF2, ATP, RNA and with UPF1 itself. Mutating these amino acids in silico showed an important effect over the native structure. Finally, we performed molecular dynamic simulations for UPF1 proteins from H. sapiens and U. maydis and the results obtained show a similar behavior and physicochemical properties for the protein in both organisms. Overall, our results indicate that the putative UPF1 identified in U. maydis shows a very similar sequence, structural organization

  20. Phytohormone Involvement in the Ustilago maydis- Zea mays Pathosystem: Relationships between Abscisic Acid and Cytokinin Levels and Strain Virulence in Infected Cob Tissue.

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    Erin N Morrison

    Full Text Available Ustilago maydis is the causative agent of common smut of corn. Early studies noted its ability to synthesize phytohormones and, more recently these growth promoting substances were confirmed as cytokinins (CKs. Cytokinins comprise a group of phytohormones commonly associated with actively dividing tissues. Lab analyses identified variation in virulence between U. maydis dikaryon and solopathogen infections of corn cob tissue. Samples from infected cob tissue were taken at sequential time points post infection and biochemical profiling was performed using high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI MS/MS. This hormone profiling revealed that there were altered levels of ABA and major CKs, with a marked reduction in CK glucosides, increases in methylthiol CKs and a particularly dramatic increase in cisZ CK forms, in U. maydis infected tissue. These changes were more pronounced in the more virulent dikaryon relative to the solopathogenic strain suggesting a role for cytokinins in moderating virulence during biotrophic infection. These findings highlight the fact that U. maydis does not simply mimic a fertilized seed but instead reprograms the host tissue. Results underscore the suitability of the Ustilago maydis- Zea mays model as a basis for investigating the control of phytohormone dynamics during biotrophic infection of plants.

  1. Phytohormone Involvement in the Ustilago maydis- Zea mays Pathosystem: Relationships between Abscisic Acid and Cytokinin Levels and Strain Virulence in Infected Cob Tissue.

    Science.gov (United States)

    Morrison, Erin N; Emery, R J Neil; Saville, Barry J

    2015-01-01

    Ustilago maydis is the causative agent of common smut of corn. Early studies noted its ability to synthesize phytohormones and, more recently these growth promoting substances were confirmed as cytokinins (CKs). Cytokinins comprise a group of phytohormones commonly associated with actively dividing tissues. Lab analyses identified variation in virulence between U. maydis dikaryon and solopathogen infections of corn cob tissue. Samples from infected cob tissue were taken at sequential time points post infection and biochemical profiling was performed using high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI MS/MS). This hormone profiling revealed that there were altered levels of ABA and major CKs, with a marked reduction in CK glucosides, increases in methylthiol CKs and a particularly dramatic increase in cisZ CK forms, in U. maydis infected tissue. These changes were more pronounced in the more virulent dikaryon relative to the solopathogenic strain suggesting a role for cytokinins in moderating virulence during biotrophic infection. These findings highlight the fact that U. maydis does not simply mimic a fertilized seed but instead reprograms the host tissue. Results underscore the suitability of the Ustilago maydis- Zea mays model as a basis for investigating the control of phytohormone dynamics during biotrophic infection of plants.

  2. The transcription factor Rbf1 is the master regulator for b-mating type controlled pathogenic development in Ustilago maydis.

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    Kai Heimel

    Full Text Available In the phytopathogenic basidiomycete Ustilago maydis, sexual and pathogenic development are tightly connected and controlled by the heterodimeric bE/bW transcription factor complex encoded by the b-mating type locus. The formation of the active bE/bW heterodimer leads to the formation of filaments, induces a G2 cell cycle arrest, and triggers pathogenicity. Here, we identify a set of 345 bE/bW responsive genes which show altered expression during these developmental changes; several of these genes are associated with cell cycle coordination, morphogenesis and pathogenicity. 90% of the genes that show altered expression upon bE/bW-activation require the zinc finger transcription factor Rbf1, one of the few factors directly regulated by the bE/bW heterodimer. Rbf1 is a novel master regulator in a multilayered network of transcription factors that facilitates the complex regulatory traits of sexual and pathogenic development.

  3. Transcriptional regulation of the genes encoding chitin and β-1,3-glucan synthases from Ustilago maydis.

    Science.gov (United States)

    Robledo-Briones, Mariana; Ruiz-Herrera, José

    2012-07-01

    Transcriptional regulation of genes encoding chitin synthases (CHS) and β-1,3-glucan synthase (GLS) from Ustilago maydis was studied. Transcript levels were measured during the growth curve of yeast and mycelial forms, in response to ionic and osmotic stress, and during infection of maize plants. Expression of the single GLS gene was constitutive. In contrast, CHS genes expression showed differences depending on environmental conditions. Transcript levels were slightly higher in the mycelial forms, the highest levels occurring at the log phase. Ionic and osmotic stress induced alterations in the expression of CHS genes, but not following a defined pattern, some genes were induced and others repressed by the tested compounds. Changes in transcripts were more apparent during the pathogenic process. At early infection stages, only CHS6 gene showed significant transcript levels, whereas at the period of tumor formation CHS7 and CHS8 genes were also were induced.

  4. The transcription factor Rbf1 is the master regulator for b-mating type controlled pathogenic development in Ustilago maydis.

    Science.gov (United States)

    Heimel, Kai; Scherer, Mario; Vranes, Miroslav; Wahl, Ramon; Pothiratana, Chetsada; Schuler, David; Vincon, Volker; Finkernagel, Florian; Flor-Parra, Ignacio; Kämper, Jörg

    2010-08-05

    In the phytopathogenic basidiomycete Ustilago maydis, sexual and pathogenic development are tightly connected and controlled by the heterodimeric bE/bW transcription factor complex encoded by the b-mating type locus. The formation of the active bE/bW heterodimer leads to the formation of filaments, induces a G2 cell cycle arrest, and triggers pathogenicity. Here, we identify a set of 345 bE/bW responsive genes which show altered expression during these developmental changes; several of these genes are associated with cell cycle coordination, morphogenesis and pathogenicity. 90% of the genes that show altered expression upon bE/bW-activation require the zinc finger transcription factor Rbf1, one of the few factors directly regulated by the bE/bW heterodimer. Rbf1 is a novel master regulator in a multilayered network of transcription factors that facilitates the complex regulatory traits of sexual and pathogenic development.

  5. Transcriptomic analysis of the dimorphic transition of Ustilago maydis induced in vitro by a change in pH.

    Science.gov (United States)

    Martínez-Soto, Domingo; Ruiz-Herrera, José

    2013-01-01

    Dimorphism is the property of fungi to grow as budding yeasts or mycelium, depending on the environmental conditions. This phenomenon is important as a model of differentiation in eukaryotic organisms, and since a large number of fungal diseases are caused by dimorphic fungi, its study is important for practical reasons. In this work, we examined the transcriptome during the dimorphic transition of the basidiomycota phytopathogenic fungus Ustilago maydis using microarrays, utilizing yeast and mycelium monomorphic mutants as controls. This way, we thereby identified 154 genes of the fungus that are specifically involved in the dimorphic transition induced by a pH change. Of these, 82 genes were up-regulated, and 72 were down-regulated. Differential categorization of these genes revealed that they mostly belonged to the classes of metabolism, cell cycle and DNA processing, transcription and protein fate, transport and cellular communication, stress, cell differentiation and biogenesis of cellular components, while a significant number of them corresponded to unclassified proteins. The data reported in this work are important for our understanding of the molecular bases of dimorphism in U. maydis, and possibly of other fungi.

  6. Transcriptomic analysis of the GCN5 gene reveals mechanisms of the epigenetic regulation of virulence and morphogenesis in Ustilago maydis.

    Science.gov (United States)

    Martínez-Soto, Domingo; González-Prieto, Juan Manuel; Ruiz-Herrera, José

    2015-09-01

    Chromatin in the eukaryotic nucleus is highly organized in the form of nucleosomes where histones wrap DNA. This structure may be altered by some chemical modifications of histones, one of them, acetylation by histone acetyltransferases (HATs) that originates relaxation of the nucleosome structure, providing access to different transcription factors and other effectors. In this way, HATs regulate cellular processes including DNA replication, and gene transcription. Previously, we isolated Ustilago maydis mutants deficient in the GCN5 HAT that are avirulent, and grow constitutively as mycelium. In this work, we proceeded to identify the genes differentially regulated by GCN5, comparing the transcriptomes of the mutant and the wild type using microarrays, to analyse the epigenetic control of virulence and morphogenesis. We identified 1203 genes, 574 positively and 629 negatively regulated in the wild type. We found that genes belonging to different categories involved in pathogenesis were downregulated in the mutant, and that genes involved in mycelial growth were negatively regulated in the wild type, offering a working hypothesis on the epigenetic control of virulence and morphogenesis of U. maydis. Interestingly, several differentially regulated genes appeared in clusters, suggesting a common regulation. Some of these belonged to pathogenesis or secondary metabolism.

  7. Molecular cloning and heterologous expression in Pichia pastoris of X-prolyl-dipeptidyl aminopeptidase from basidiomycete Ustilago maydis.

    Science.gov (United States)

    Juárez-Montiel, Margarita; Ibarra, J Antonio; Chávez-Camarillo, Griselda; Hernández-Rodríguez, César; Villa-Tanaca, Lourdes

    2014-03-01

    Dipeptidyl aminopeptidases are enzymes involved in the posttranslational control of bioactive peptides. Here we identified the gene dapUm in Ustilago maydis by homology with other fungal dipeptidyl aminopeptidases. Analysis of the dapUm-deduced amino acid sequence indicated that it encodes for membrane-type serine protease with a characteristic prolyl oligopeptidase catalytic motif triad: Ser, Asp, His. In order to overexpress the DapUm, the gene encoding for it was cloned and transformed into Pichia. Using this system, we observed a ∼ 125-kDa recombinant protein with an optimal enzymatic activity at pH 6.0 and at 40 °C for the Ala-Pro-p-nitroanilide substrate and an experimental pH of 6.9. U. maydis DapUm was specifically inhibited by phenylmethylsulfonyl fluoride and Pefabloc, confirming the presence of a serine residue in the active site. To our knowledge, this study is the first report on the cloning and expression of a DPP IV dipeptidyl aminopeptidase from a basidiomycete organism. Moreover, the use of recombinant DapUm will allow us to further study and characterize this enzyme, in addition to testing chemical compounds for pharmaceutical purposes.

  8. Analysis of the regulation of the Ustilago maydis proteome by dimorphism, pH or MAPK and GCN5 genes.

    Science.gov (United States)

    Martínez-Salgado, José L; León-Ramírez, Claudia G; Pacheco, Alberto Barrera; Ruiz-Herrera, José; de la Rosa, Ana P Barba

    2013-02-21

    Ustilago maydis is a dimorphic corn pathogenic basidiomycota whose haploid cells grow in yeast form at pH7, while at pH3 they grow in the mycelial form. Two-dimensional gel electrophoresis (2-DE) coupled with LC-ESI/MS-MS was used to analyze the differential accumulation of proteins in yeast against mycelial morphologies. 2-DE maps were obtained in the pH range of 5-8 and 404 total protein spots were separated. From these, 43 were differentially accumulated when comparing strains FB2wt, constitutive yeast CL211, and constitutive mycelial GP25 growing at pH7 against pH3. Differentially accumulated proteins in response to pH are related with defense against reactive oxygen species or toxic compounds. Up-accumulation of CipC and down-accumulation of Hmp1 were specifically related with mycelial growth. Changes in proteins that were affected by mutation in the gene encoding the adaptor of a MAPK pathway (CL211 strain) were UM521* and transcription factors Btf3, Sol1 and Sti1. Mutation of GCN5 (GP25 strain) affected the accumulation of Rps19-ribosomal protein, Mge1-heath shock protein, and Lpd1-dihydrolipoamide dehydrogenase. Our results complement the information about the genes and proteins related with the dimorphic transition in U. maydis and changes in proteins affected by mutations in a MAPK pathway and GCN5 gene.

  9. The Ustilago maydis Nit2 homolog regulates nitrogen utilization and is required for efficient induction of filamentous growth.

    Science.gov (United States)

    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.

  10. The tryptophan aminotransferase Tam1 catalyses the single biosynthetic step for tryptophan-dependent pigment synthesis in Ustilago maydis.

    Science.gov (United States)

    Zuther, Katja; Mayser, Peter; Hettwer, Ursula; Wu, Wenying; Spiteller, Peter; Kindler, Bernhard L J; Karlovsky, Petr; Basse, Christoph W; Schirawski, Jan

    2008-04-01

    Tryptophan is a precursor for many biologically active secondary metabolites. We have investigated the origin of indole pigments first described in the pityriasis versicolor-associated fungus Malassezia furfur. Some of the identified indole pigments have properties potentially explaining characteristics of the disease. As M. furfur is not amenable to genetic manipulation, we used Ustilago maydis to investigate the pathway leading to pigment production from tryptophan. We show by high-performance liquid chromatography, mass spectrometry and nuclear magnetic resonance analysis that the compounds produced by U. maydis include those putatively involved in the etiology of pityriasis versicolor. Using a reverse genetics approach, we demonstrate that the tryptophan aminotransferase Tam1 catalyses pigment biosynthesis by conversion of tryptophan into indolepyruvate. A forward genetics approach led to the identification of mutants incapable of producing the pigments. These mutants were affected in the sir1 gene, presumably encoding a sulphite reductase. In vitro experiments with purified Tam1 showed that 2-oxo 4-methylthio butanoate serves as a substrate linking tryptophan deamination to sulphur metabolism. We provide the first direct evidence that these indole pigments form spontaneously from indolepyruvate and tryptophan without any enzymatic activity. This suggests that compounds with a proposed function in M. furfur-associated disease consist of indolepyruvate-derived spontaneously generated metabolic by-products.

  11. Post-genomic analyses of fungal lignocellulosic biomass degradation reveal the unexpected potential of the plant pathogen Ustilago maydis

    Directory of Open Access Journals (Sweden)

    Couturier Marie

    2012-02-01

    Full Text Available Abstract Background Filamentous fungi are potent biomass degraders due to their ability to thrive in ligno(hemicellulose-rich environments. During the last decade, fungal genome sequencing initiatives have yielded abundant information on the genes that are putatively involved in lignocellulose degradation. At present, additional experimental studies are essential to provide insights into the fungal secreted enzymatic pools involved in lignocellulose degradation. Results In this study, we performed a wide analysis of 20 filamentous fungi for which genomic data are available to investigate their biomass-hydrolysis potential. A comparison of fungal genomes and secretomes using enzyme activity profiling revealed discrepancies in carbohydrate active enzymes (CAZymes sets dedicated to plant cell wall. Investigation of the contribution made by each secretome to the saccharification of wheat straw demonstrated that most of them individually supplemented the industrial Trichoderma reesei CL847 enzymatic cocktail. Unexpectedly, the most striking effect was obtained with the phytopathogen Ustilago maydis that improved the release of total sugars by 57% and of glucose by 22%. Proteomic analyses of the best-performing secretomes indicated a specific enzymatic mechanism of U. maydis that is likely to involve oxido-reductases and hemicellulases. Conclusion This study provides insight into the lignocellulose-degradation mechanisms by filamentous fungi and allows for the identification of a number of enzymes that are potentially useful to further improve the industrial lignocellulose bioconversion process.

  12. Physical and genetic interaction between ammonium transporters and the signaling protein Rho1 in the plant pathogen Ustilago maydis.

    Science.gov (United States)

    Paul, Jinny A; Barati, Michelle T; Cooper, Michael; Perlin, Michael H

    2014-10-01

    Dimorphic transitions between yeast-like and filamentous forms occur in many fungi and are often associated with pathogenesis. One of the cues for such a dimorphic switch is the availability of nutrients. Under conditions of nitrogen limitation, fungal cells (such as those of Saccharomyces cerevisiae and Ustilago maydis) switch from budding to pseudohyphal or filamentous growth. Ammonium transporters (AMTs) are responsible for uptake and, in some cases, for sensing the availability of ammonium, a preferred nitrogen source. Homodimer and/or heterodimer formation may be required for regulating the activity of the AMTs. To investigate the potential interactions of Ump1 and Ump2, the AMTs of the maize pathogen U. maydis, we first used the split-ubiquitin system, followed by a modified split-YFP (yellow fluorescent protein) system, to validate the interactions in vivo. This analysis showed the formation of homo- and hetero-oligomers by Ump1 and Ump2. We also demonstrated the interaction of the high-affinity ammonium transporter, Ump2, with the Rho1 GTPase, a central protein in signaling, with roles in controlling polarized growth. This is the first demonstration in eukaryotes of the physical interaction in vivo of an ammonium transporter with the signaling protein Rho1. Moreover, the Ump proteins interact with Rho1 during the growth of cells in low ammonium concentrations, a condition required for the expression of the Umps. Based on these results and the genetic evidence for the interaction of Ump2 with both Rho1 and Rac1, another small GTPase, we propose a model for the role of these interactions in controlling filamentation, a fundamental aspect of development and pathogenesis in U. maydis.

  13. A rapid and efficient method for assessing pathogenicity of ustilago maydis on maize and teosinte lines.

    Science.gov (United States)

    Chavan, Suchitra; Smith, Shavannor M

    2014-01-03

    Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.(1) Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut(2). However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually(3-5). Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants. Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

  14. Protein glycosylation in the phytopathogen Ustilago maydis: From core oligosaccharide synthesis to the ER glycoprotein quality control system, a genomic analysis.

    Science.gov (United States)

    Fernández-Alvarez, Alfonso; Elías-Villalobos, Alberto; Ibeas, José I

    2010-09-01

    The corn smut fungus Ustilago maydis has, over recent decades, become established as a robust pathogenic model for studying fungi-plant relationships. This use of U. maydis can be attributed to its biotrophic host interaction, easy culture and genetic manipulation in the laboratory, and the severe disease symptoms it induces in infected maize. Recent studies have shown that normal protein glycosylation is essential for pathogenic development, but dispensable for the saprophytic growth or mating. Given the relevance of protein glycosylation for U. maydis virulence, and consequently its role in the plant pathogenesis, here we review the main actors and events implicated in protein glycosylation. Furthermore, we describe the results of an in silico search, where we identify all the conserved members of the N- and O-glycosylation pathways in U. maydis at each stage: core oligosaccharide synthesis, addition of the core oligosaccharide to nascent target proteins, maturation and extension of the core oligosaccharide, and the quality control system used by the cell to avoid the presence of unfolded glycoproteins. Finally, we discuss how these genes could affect U. maydis virulence and their biotechnological implications.

  15. The SPF27 homologue Num1 connects splicing and kinesin 1-dependent cytoplasmic trafficking in Ustilago maydis.

    Science.gov (United States)

    Kellner, Nikola; Heimel, Kai; Obhof, Theresa; Finkernagel, Florian; Kämper, Jörg

    2014-01-01

    The conserved NineTeen protein complex (NTC) is an integral subunit of the spliceosome and required for intron removal during pre-mRNA splicing. The complex associates with the spliceosome and participates in the regulation of conformational changes of core spliceosomal components, stabilizing RNA-RNA- as well as RNA-protein interactions. In addition, the NTC is involved in cell cycle checkpoint control, response to DNA damage, as well as formation and export of mRNP-particles. We have identified the Num1 protein as the homologue of SPF27, one of NTC core components, in the basidiomycetous fungus Ustilago maydis. Num1 is required for polarized growth of the fungal hyphae, and, in line with the described NTC functions, the num1 mutation affects the cell cycle and cell division. The num1 deletion influences splicing in U. maydis on a global scale, as RNA-Seq analysis revealed increased intron retention rates. Surprisingly, we identified in a screen for Num1 interacting proteins not only NTC core components as Prp19 and Cef1, but several proteins with putative functions during vesicle-mediated transport processes. Among others, Num1 interacts with the motor protein Kin1 in the cytoplasm. Similar phenotypes with respect to filamentous and polar growth, vacuolar morphology, as well as the motility of early endosomes corroborate the genetic interaction between Num1 and Kin1. Our data implicate a previously unidentified connection between a component of the splicing machinery and cytoplasmic transport processes. As the num1 deletion also affects cytoplasmic mRNA transport, the protein may constitute a novel functional interconnection between the two disparate processes of splicing and trafficking.

  16. The SPF27 homologue Num1 connects splicing and kinesin 1-dependent cytoplasmic trafficking in Ustilago maydis.

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    Nikola Kellner

    2014-01-01

    Full Text Available The conserved NineTeen protein complex (NTC is an integral subunit of the spliceosome and required for intron removal during pre-mRNA splicing. The complex associates with the spliceosome and participates in the regulation of conformational changes of core spliceosomal components, stabilizing RNA-RNA- as well as RNA-protein interactions. In addition, the NTC is involved in cell cycle checkpoint control, response to DNA damage, as well as formation and export of mRNP-particles. We have identified the Num1 protein as the homologue of SPF27, one of NTC core components, in the basidiomycetous fungus Ustilago maydis. Num1 is required for polarized growth of the fungal hyphae, and, in line with the described NTC functions, the num1 mutation affects the cell cycle and cell division. The num1 deletion influences splicing in U. maydis on a global scale, as RNA-Seq analysis revealed increased intron retention rates. Surprisingly, we identified in a screen for Num1 interacting proteins not only NTC core components as Prp19 and Cef1, but several proteins with putative functions during vesicle-mediated transport processes. Among others, Num1 interacts with the motor protein Kin1 in the cytoplasm. Similar phenotypes with respect to filamentous and polar growth, vacuolar morphology, as well as the motility of early endosomes corroborate the genetic interaction between Num1 and Kin1. Our data implicate a previously unidentified connection between a component of the splicing machinery and cytoplasmic transport processes. As the num1 deletion also affects cytoplasmic mRNA transport, the protein may constitute a novel functional interconnection between the two disparate processes of splicing and trafficking.

  17. Patterns of variation at Ustilago maydis virulence clusters 2A and 19A largely reflect the demographic history of its populations.

    Science.gov (United States)

    Kellner, Ronny; Hanschke, Christian; Begerow, Dominik

    2014-01-01

    The maintenance of an intimate interaction between plant-biotrophic fungi and their hosts over evolutionary times involves strong selection and adaptative evolution of virulence-related genes. The highly specialised maize pathogen Ustilago maydis is assigned with a high evolutionary capability to overcome host resistances due to its high rates of sexual recombination, large population sizes and long distance dispersal. Unlike most studied fungus-plant interactions, the U. maydis - Zea mays pathosystem lacks a typical gene-for-gene interaction. It exerts a large set of secreted fungal virulence factors that are mostly organised in gene clusters. Their contribution to virulence has been experimentally demonstrated but their genetic diversity within U. maydis remains poorly understood. Here, we report on the intraspecific diversity of 34 potential virulence factor genes of U. maydis. We analysed their sequence polymorphisms in 17 isolates of U. maydis from Europe, North and Latin America. We focused on gene cluster 2A, associated with virulence attenuation, cluster 19A that is crucial for virulence, and the cluster-independent effector gene pep1. Although higher compared to four house-keeping genes, the overall levels of intraspecific genetic variation of virulence clusters 2A and 19A, and pep1 are remarkably low and commensurate to the levels of 14 studied non-virulence genes. In addition, each gene is present in all studied isolates and synteny in cluster 2A is conserved. Furthermore, 7 out of 34 virulence genes contain either no polymorphisms or only synonymous substitutions among all isolates. However, genetic variation of clusters 2A and 19A each resolve the large scale population structure of U. maydis indicating subpopulations with decreased gene flow. Hence, the genetic diversity of these virulence-related genes largely reflect the demographic history of U. maydis populations.

  18. Patterns of variation at Ustilago maydis virulence clusters 2A and 19A largely reflect the demographic history of its populations.

    Directory of Open Access Journals (Sweden)

    Ronny Kellner

    Full Text Available The maintenance of an intimate interaction between plant-biotrophic fungi and their hosts over evolutionary times involves strong selection and adaptative evolution of virulence-related genes. The highly specialised maize pathogen Ustilago maydis is assigned with a high evolutionary capability to overcome host resistances due to its high rates of sexual recombination, large population sizes and long distance dispersal. Unlike most studied fungus-plant interactions, the U. maydis - Zea mays pathosystem lacks a typical gene-for-gene interaction. It exerts a large set of secreted fungal virulence factors that are mostly organised in gene clusters. Their contribution to virulence has been experimentally demonstrated but their genetic diversity within U. maydis remains poorly understood. Here, we report on the intraspecific diversity of 34 potential virulence factor genes of U. maydis. We analysed their sequence polymorphisms in 17 isolates of U. maydis from Europe, North and Latin America. We focused on gene cluster 2A, associated with virulence attenuation, cluster 19A that is crucial for virulence, and the cluster-independent effector gene pep1. Although higher compared to four house-keeping genes, the overall levels of intraspecific genetic variation of virulence clusters 2A and 19A, and pep1 are remarkably low and commensurate to the levels of 14 studied non-virulence genes. In addition, each gene is present in all studied isolates and synteny in cluster 2A is conserved. Furthermore, 7 out of 34 virulence genes contain either no polymorphisms or only synonymous substitutions among all isolates. However, genetic variation of clusters 2A and 19A each resolve the large scale population structure of U. maydis indicating subpopulations with decreased gene flow. Hence, the genetic diversity of these virulence-related genes largely reflect the demographic history of U. maydis populations.

  19. Unfolded Protein Response (UPR Regulator Cib1 Controls Expression of Genes Encoding Secreted Virulence Factors in Ustilago maydis.

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    Martin Hampel

    Full Text Available The unfolded protein response (UPR, a conserved eukaryotic signaling pathway to ensure protein homeostasis in the endoplasmic reticulum (ER, coordinates biotrophic development in the corn smut fungus Ustilago maydis. Exact timing of UPR activation is required for virulence and presumably connected to the elevated expression of secreted effector proteins during infection of the host plant Zea mays. In the baker's yeast Saccharomyces cerevisiae, expression of UPR target genes is induced upon binding of the central regulator Hac1 to unfolded protein response elements (UPREs in their promoters. While a role of the UPR in effector secretion has been described previously, we investigated a potential UPR-dependent regulation of genes encoding secreted effector proteins. In silico prediction of UPREs in promoter regions identified the previously characterized effector genes pit2 and tin1-1, as bona fide UPR target genes. Furthermore, direct binding of the Hac1-homolog Cib1 to the UPRE containing promoter fragments of both genes was confirmed by quantitative chromatin immunoprecipitation (qChIP analysis. Targeted deletion of the UPRE abolished Cib1-dependent expression of pit2 and significantly affected virulence. Furthermore, ER stress strongly increased Pit2 expression and secretion. This study expands the role of the UPR as a signal hub in fungal virulence and illustrates, how biotrophic fungi can coordinate cellular physiology, development and regulation of secreted virulence factors.

  20. Biological Characteristics of Ustilago maydis%玉米瘤黑粉病菌生物学特性研究

    Institute of Scientific and Technical Information of China (English)

    肖淑芹; 姜晓颖; 黄伟东; 张悦; 魏二琴; 薛春生; 石洁

    2011-01-01

    Conditions of teliospore germination and dicaryophyte growth of Ustilago maydis were studied. The results showed that the optimum temperature, pH, carbon source and nitrogen source for teliospore germination were 30℃, 3, sucrose and Cysteine, the optimum temperature, pH, carbon source and nitrogen source for dicaryophyte growth were 30℃, 8, glucose and Aspartic acid. Teliospore germination and dicaryophyte growth were not significantly effective on different light conditions.%对影响玉蜀黍黑粉菌冬孢子萌发和双核菌丝生长的生物学特性进行研究.结果表明,冬孢子萌发最适温度为30℃,pH为3,碳源为蔗糖、麦芽糖和葡萄糖,氮源为半胱氨酸、天冬氨酸和甘氨酸;双核菌丝生长的最适温度为30℃,pH为8,碳源为葡萄糖和蔗糖,氮源为天冬氨酸、半胱氨酸和甘氨酸.光照条件对冬孢子萌发和菌丝生长的影响不显著.

  1. The UmGcn5 gene encoding histone acetyltransferase from Ustilago maydis is involved in dimorphism and virulence.

    Science.gov (United States)

    González-Prieto, Juan Manuel; Rosas-Quijano, Raymundo; Domínguez, Angel; Ruiz-Herrera, José

    2014-10-01

    We isolated a gene encoding a histone acetyltransferase from Ustilago maydis (DC.) Cda., which is orthologous to the Saccharomyces cerevisiae GCN5 gene. The gene was isolated from genomic clones identified by their specific hybridization to a gene fragment obtained by the polymerase chain reaction (PCR). This gene (Umgcn5; um05168) contains an open reading frame (ORF) of 1421bp that encodes a putative protein of 473 amino acids with a Mr. of 52.6kDa. The protein exhibits a high degree of homology with histone acetyltransferases from different organisms. Null a2b2 ΔUmgcn5 mutants were constructed by substitution of the region encoding the catalytic site with a hygromycin B resistance cassette. Null a1b1 ΔUmgcn5 mutants were isolated from genetic crosses of a2b2 ΔUmgcn5 and a1b1 wild-type strains in maize. Mutants displayed a slight reduction in growth rate under different conditions, and were more sensitive than the wild type to stress conditions, but more important, they grew as long mycelial cells, and formed fuzz-like colonies under all conditions where wild-type strains grew in the yeast-like morphology and formed smooth colonies. This phenotype was not reverted by cAMP addition. Mutants were not virulent to maize plants, and were unable to form teliospores. These phenotypic alterations of the mutants were reverted by their transformation with the wild-type gene.

  2. A novel high-affinity sucrose transporter is required for virulence of the plant pathogen Ustilago maydis.

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    Ramon Wahl

    2010-02-01

    Full Text Available Plant pathogenic fungi cause massive yield losses and affect both quality and safety of food and feed produced from infected plants. The main objective of plant pathogenic fungi is to get access to the organic carbon sources of their carbon-autotrophic hosts. However, the chemical nature of the carbon source(s and the mode of uptake are largely unknown. Here, we present a novel, plasma membrane-localized sucrose transporter (Srt1 from the corn smut fungus Ustilago maydis and its characterization as a fungal virulence factor. Srt1 has an unusually high substrate affinity, is absolutely sucrose specific, and allows the direct utilization of sucrose at the plant/fungal interface without extracellular hydrolysis and, thus, without the production of extracellular monosaccharides known to elicit plant immune responses. srt1 is expressed exclusively during infection, and its deletion strongly reduces fungal virulence. This emphasizes the central role of this protein both for efficient carbon supply and for avoidance of apoplastic signals potentially recognized by the host.

  3. Unfolded Protein Response (UPR) Regulator Cib1 Controls Expression of Genes Encoding Secreted Virulence Factors in Ustilago maydis.

    Science.gov (United States)

    Hampel, Martin; Jakobi, Mareike; Schmitz, Lara; Meyer, Ute; Finkernagel, Florian; Doehlemann, Gunther; Heimel, Kai

    2016-01-01

    The unfolded protein response (UPR), a conserved eukaryotic signaling pathway to ensure protein homeostasis in the endoplasmic reticulum (ER), coordinates biotrophic development in the corn smut fungus Ustilago maydis. Exact timing of UPR activation is required for virulence and presumably connected to the elevated expression of secreted effector proteins during infection of the host plant Zea mays. In the baker's yeast Saccharomyces cerevisiae, expression of UPR target genes is induced upon binding of the central regulator Hac1 to unfolded protein response elements (UPREs) in their promoters. While a role of the UPR in effector secretion has been described previously, we investigated a potential UPR-dependent regulation of genes encoding secreted effector proteins. In silico prediction of UPREs in promoter regions identified the previously characterized effector genes pit2 and tin1-1, as bona fide UPR target genes. Furthermore, direct binding of the Hac1-homolog Cib1 to the UPRE containing promoter fragments of both genes was confirmed by quantitative chromatin immunoprecipitation (qChIP) analysis. Targeted deletion of the UPRE abolished Cib1-dependent expression of pit2 and significantly affected virulence. Furthermore, ER stress strongly increased Pit2 expression and secretion. This study expands the role of the UPR as a signal hub in fungal virulence and illustrates, how biotrophic fungi can coordinate cellular physiology, development and regulation of secreted virulence factors.

  4. The AGC Ser/Thr kinase Aga1 is essential for appressorium formation and maintenance of the actin cytoskeleton in the smut fungus Ustilago maydis.

    Science.gov (United States)

    Berndt, Patrick; Lanver, Daniel; Kahmann, Regine

    2010-12-01

    On the plant surface the dimorphic fungus Ustilago maydis switches from budding to hyphal growth and differentiates appressoria. To get more insight into these highly regulated processes we report on the role of a conserved Ser/Thr kinase of the AGC kinase family, Aga1. U. maydis Aga1 could functionally replace Ypk1p in Saccharomyces cerevisiae. aga1 deletion mutants were affected in growth, cell wall integrity, mating as well as the ability to form appressoria and showed defects in actin organization and actin-dependent endocytosis. With respect to appressorium formation and endocytosis, the aga1 deletion phenotype could be mimicked by inhibiting the formation of actin filaments with Latrunculin A. These data suggest a critical role of Aga1 in F-actin organization during the morphological changes accompanying the development of appressoria.

  5. Emergence and prevention measures of Ustilago maydis in sweet corn seed production%制种田玉米瘤黑粉病的发生及防治

    Institute of Scientific and Technical Information of China (English)

    王辉; 何云龙; 谢立剑

    2003-01-01

    1 病原、寄主植物及分布玉米瘤黑粉病由瘤黑粉菌( Ustilago maydis)引起发病,属担子菌亚门,常为害玉米叶、秆、雄穗和果穗等部位的幼嫩组织,产生大小不一的病瘤,是我国玉米上分布最广的主要病害之一,北方发生较为普遍而严重.

  6. Utilizing virus-induced gene silencing for the functional characterization of maize genes during infection with the fungal pathogen Ustilago maydis.

    Science.gov (United States)

    van der Linde, Karina; Doehlemann, Gunther

    2013-01-01

    While in dicotyledonous plants virus-induced gene silencing (VIGS) is well established to study plant-pathogen interaction, in monocots only few examples of efficient VIGS have been reported so far. One of the available systems is based on the brome mosaic virus (BMV) which allows gene silencing in different cereals including barley (Hordeum vulgare), wheat (Triticum aestivum), and maize (Zea mays).Infection of maize plants by the corn smut fungus Ustilago maydis leads to the formation of large tumors on stem, leaves, and inflorescences. During this biotrophic interaction, plant defense responses are actively suppressed by the pathogen, and previous transcriptome analyses of infected maize plants showed comprehensive and stage-specific changes in host gene expression during disease progression.To identify maize genes that are functionally involved in the interaction with U. maydis, we adapted a VIGS system based on the Brome mosaic virus (BMV) to maize at conditions that allow successful U. maydis infection of BMV pre-infected maize plants. This setup enables quantification of VIGS and its impact on U. maydis infection using a quantitative real-time PCR (q(RT)-PCR)-based readout.

  7. Indole-3-acetic acid (IAA) biosynthesis in the smut fungus Ustilago maydis and its relevance for increased IAA levels in infected tissue and host tumour formation.

    Science.gov (United States)

    Reineke, Gavin; Heinze, Bernadette; Schirawski, Jan; Buettner, Hermann; Kahmann, Regine; Basse, Christoph W

    2008-05-01

    Infection of maize (Zea mays) plants with the smut fungus Ustilago maydis is characterized by excessive host tumour formation. U. maydis is able to produce indole-3-acetic acid (IAA) efficiently from tryptophan. To assess a possible connection to the induction of host tumours, we investigated the pathways leading to fungal IAA biosynthesis. Besides the previously identified iad1 gene, we identified a second indole-3-acetaldehyde dehydrogenase gene, iad2. Deltaiad1Deltaiad2 mutants were blocked in the conversion of both indole-3-acetaldehyde and tryptamine to IAA, although the reduction in IAA formation from tryptophan was not significantly different from Deltaiad1 mutants. To assess an influence of indole-3-pyruvic acid on IAA formation, we deleted the aromatic amino acid aminotransferase genes tam1 and tam2 in Deltaiad1Deltaiad2 mutants. This revealed a further reduction in IAA levels by five- and tenfold in mutant strains harbouring theDeltatam1 andDeltatam1Deltatam2 deletions, respectively. This illustrates that indole-3-pyruvic acid serves as an efficient precursor for IAA formation in U. maydis. Interestingly, the rise in host IAA levels upon U. maydis infection was significantly reduced in tissue infected with Deltaiad1Deltaiad2Deltatam1 orDeltaiad1Deltaiad2Deltatam1Deltatam2 mutants, whereas induction of tumours was not compromised. Together, these results indicate that fungal IAA production critically contributes to IAA levels in infected tissue, but this is apparently not important for triggering host tumour formation.

  8. Mycosarcoma (Ustilaginaceae), a resurrected generic name for corn smut (Ustilago maydis) and its close relatives with hypertrophied, tubular sori

    NARCIS (Netherlands)

    McTaggart, Alistair R; Shivas, Roger G; Boekhout, Teun; Oberwinkler, Franz; Vánky, Kálmán; Pennycook, Shaun R; Begerow, Dominik

    2016-01-01

    Ustilago is a polyphyletic genus of smut fungi found mainly on Poaceae. The development of a taxonomy that reflects phylogeny requires subdivision of Ustilago into smaller monophyletic genera. Several separate systematic analyses have determined that Macalpinomyces mackinlayi, M. tubiformis, Tolypos

  9. Mre11 and Blm-Dependent Formation of ALT-Like Telomeres in Ku-Deficient Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Eun Young Yu

    2015-10-01

    Full Text Available A subset of human cancer cells uses a specialized, aberrant recombination pathway known as ALT to maintain telomeres, which in these cells are characterized by complex aberrations including length heterogeneity, high levels of unpaired C-strand, and accumulation of extra-chromosomal telomere repeats (ECTR. These phenotypes have not been recapitulated in any standard budding or fission yeast mutant. We found that eliminating Ku70 or Ku80 in the yeast-like fungus Ustilago maydis results initially in all the characteristic telomere aberrations of ALT cancer cells, including C-circles, a highly specific marker of ALT. Subsequently the ku mutants experience permanent G2 cell cycle arrest, accompanied by loss of telomere repeats from chromosome ends and even more drastic accumulation of very short ECTRs (vsECTRs. The deletion of atr1 or chk1 rescued the lethality of the ku mutant, and "trapped" the telomere aberrations in the early ALT-like stage. Telomere abnormalities are telomerase-independent, but dramatically suppressed by deletion of mre11 or blm, suggesting major roles for these factors in the induction of the ALT pathway. In contrast, removal of other DNA damage response and repair factors such as Rad51 has disparate effects on the ALT phenotypes, suggesting that these factors process ALT intermediates or products. Notably, the antagonism of Ku and Mre11 in the induction of ALT is reminiscent of their roles in DSB resection, in which Blm is also known to play a key role. We suggest that an aberrant resection reaction may constitute an early trigger for ALT telomeres, and that the outcomes of ALT are distinct from DSB because of the unique telomere nucleoprotein structure.

  10. Mre11 and Blm-Dependent Formation of ALT-Like Telomeres in Ku-Deficient Ustilago maydis.

    Science.gov (United States)

    Yu, Eun Young; Pérez-Martín, José; Holloman, William K; Lue, Neal F

    2015-10-01

    A subset of human cancer cells uses a specialized, aberrant recombination pathway known as ALT to maintain telomeres, which in these cells are characterized by complex aberrations including length heterogeneity, high levels of unpaired C-strand, and accumulation of extra-chromosomal telomere repeats (ECTR). These phenotypes have not been recapitulated in any standard budding or fission yeast mutant. We found that eliminating Ku70 or Ku80 in the yeast-like fungus Ustilago maydis results initially in all the characteristic telomere aberrations of ALT cancer cells, including C-circles, a highly specific marker of ALT. Subsequently the ku mutants experience permanent G2 cell cycle arrest, accompanied by loss of telomere repeats from chromosome ends and even more drastic accumulation of very short ECTRs (vsECTRs). The deletion of atr1 or chk1 rescued the lethality of the ku mutant, and "trapped" the telomere aberrations in the early ALT-like stage. Telomere abnormalities are telomerase-independent, but dramatically suppressed by deletion of mre11 or blm, suggesting major roles for these factors in the induction of the ALT pathway. In contrast, removal of other DNA damage response and repair factors such as Rad51 has disparate effects on the ALT phenotypes, suggesting that these factors process ALT intermediates or products. Notably, the antagonism of Ku and Mre11 in the induction of ALT is reminiscent of their roles in DSB resection, in which Blm is also known to play a key role. We suggest that an aberrant resection reaction may constitute an early trigger for ALT telomeres, and that the outcomes of ALT are distinct from DSB because of the unique telomere nucleoprotein structure.

  11. Growth at high pH and sodium and potassium tolerance in media above the cytoplasmic pH depend on ENA ATPases in Ustilago maydis.

    Science.gov (United States)

    Benito, Begoña; Garciadeblás, Blanca; Pérez-Martín, José; Rodríguez-Navarro, Alonso

    2009-06-01

    Potassium and Na(+) effluxes across the plasma membrane are crucial processes for the ionic homeostasis of cells. In fungal cells, these effluxes are mediated by cation/H(+) antiporters and ENA ATPases. We have cloned and studied the functions of the two ENA ATPases of Ustilago maydis, U. maydis Ena1 (UmEna1) and UmEna2. UmEna1 is a typical K(+) or Na(+) efflux ATPase whose function is indispensable for growth at pH 9.0 and for even modest Na(+) or K(+) tolerances above pH 8.0. UmEna1 locates to the plasma membrane and has the characteristics of the low-Na(+)/K(+)-discrimination ENA ATPases. However, it still protects U. maydis cells in high-Na(+) media because Na(+) showed a low cytoplasmic toxicity. The UmEna2 ATPase is phylogenetically distant from UmEna1 and is located mainly at the endoplasmic reticulum. The function of UmEna2 is not clear, but we found that it shares several similarities with Neurospora crassa ENA2, which suggests that endomembrane ENA ATPases may exist in many fungi. The expression of ena1 and ena2 transcripts in U. maydis was enhanced at high pH and at high K(+) and Na(+) concentrations. We discuss that there are two modes of Na(+) tolerance in fungi: the high-Na(+)-content mode, involving ENA ATPases with low Na(+)/K(+) discrimination, as described here for U. maydis, and the low-Na(+)-content mode, involving Na(+)-specific ENA ATPases, as in Neurospora crassa.

  12. Lipid, membrane, and mitochondrial characteristics of Ustilago maydis following exposure to ergosterol biosynthesis inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Waterfield, W.F. III

    1986-01-01

    Pencoazole at 0.5 ..mu..g/ml inhibited ergosterol biosynthesis in U. maydis. Polar lipids of sporidia grown with 0.5 ..mu..g/ml penconazole for 7.5 or 22 hr or 1.0 ..mu..g/ml fenarimol for 7.5 hr contained more 18:2 than 18:1 fatty acids. There was usually more 18:1 than 18:2 fatty acids in polar lipids of untreated sporidia but this ratio was influenced by culture cell density. The high 18:2 to 18:1 ratio in the polar lipids from penconazole grown cells was unaffected by cell density. There was an increase in free fatty acids and these were enriched with 18:2 members in cells grown with 0.5 ..mu..g/ml penconazole for 22 hr. Unsaturation of triglycerides fatty acids did not differ appreciably from that of untreated sporidia. Untreated WT U. maydis protoplasts lysed more slowly in 0.3 M sorbitol than those prepared from WT sporidia grown for 16 hr with 1.0 ..mu..g/ml penconazole or 2.0 ..mu..g/ml fenarimol or from untreated erg-40 sporidia. Protoplasts were more permeable to crystal violet than were those from untreated WT sporidia. Mitochondria from untreated WT sporidia oxidizing pyruvate plus malate or succinate yielded higher ADP/O rations than mitochondria from erg-40 or penconazole grown WT sporidia. The mitochondrial ATPase of control cells had a Km of 0.8 mM ATP whereas the mitochondrial ATPase of penconazole grown WT and erg-40 had a Km value of 3.7 and 3.2 mM ATP, respectively. When the mitochondrial catalytic subunit of the ATPase from these mitochondria were solubilized, the Km did not differ. These studies suggest that changes in sterols and membrane fatty acids resulting from treatments with EBI fungicides cause increased membrane fluidity which affects membrane stability, permeability and activity of the mitochondrial ATPase.

  13. Deletion of the Ustilago maydis ortholog of the Aspergillus sporulation regulator medA affects mating and virulence through pheromone response.

    Science.gov (United States)

    Chacko, Nadia; Gold, Scott

    2012-06-01

    Mating of compatible haploid cells of Ustilago maydis is essential for infection and disease development in the host. For mating and subsequent filamentous growth and pathogenicity, the transcription factor, prf1 is necessary. Prf1 is in turn regulated by the cAMP and MAPK pathways and other regulators like rop1 and hap1. Here we describe the identification of another putative Prf1 regulator, med1, the ortholog of the Aspergillus nidulans medusa (medA) transcription factor and show that it is required for mating and full virulence in U. maydis. med1 deletion mutants show both pre- and post-mating defects and are unresponsive to external pheromone. The expression of prf1 is down-regulated in Δmed1 compared to the wild type, suggesting that med1 is upstream of prf1. Additionally, indicative of a role in secondary metabolism regulation, deletion of the med1 gene de-represses the production of glycolipids in U. maydis.

  14. Transcriptome Analysis of a Ustilago maydis ust1 Deletion Mutant Uncovers Involvement of Laccase and Polyketide Synthase Genes in Spore Development.

    Science.gov (United States)

    Islamovic, Emir; García-Pedrajas, María D; Chacko, Nadia; Andrews, David L; Covert, Sarah F; Gold, Scott E

    2015-01-01

    Ustilago maydis, causal agent of corn smut disease, is a dimorphic fungus alternating between a saprobic budding haploid and an obligate pathogenic filamentous dikaryon. Maize responds to U. maydis colonization by producing tumorous structures, and only within these does the fungus sporulate, producing melanized sexual teliospores. Previously we identified Ust1, an APSES (Asm1p, Phd1p, Sok2p, Efg1p, and StuAp) transcription factor, whose deletion led to filamentous haploid growth and the production of highly pigmented teliospore-like structures in culture. In this study, we analyzed the transcriptome of a ust1 deletion mutant and functionally characterized two highly upregulated genes with potential roles in melanin biosynthesis: um05361, encoding a putative laccase (lac1), and um06414, encoding a polyketide synthase (pks1). The Δlac1 mutant strains showed dramatically reduced virulence on maize seedlings and fewer, less-pigmented teliospores in adult plants. The Δpks1 mutant was unaffected in seedling virulence but adult plant tumors generated hyaline, nonmelanized teliospores. Thus, whereas pks1 appeared to be restricted to the synthesis of melanin, lac1 showed a broader role in virulence. In conclusion, the ust1 deletion mutant provided an in vitro model for sporulation in U. maydis, and functional analysis supports the efficacy of this in vitro mutant analysis for identification of genes involved in in planta teliosporogenesis.

  15. MRN- and 9-1-1-Independent Activation of the ATR-Chk1 Pathway during the Induction of the Virulence Program in the Phytopathogen Ustilago maydis.

    Science.gov (United States)

    Tenorio-Gómez, María; de Sena-Tomás, Carmen; Pérez-Martín, Jose

    2015-01-01

    DNA damage response (DDR) leads to DNA repair, and depending on the extent of the damage, to further events, including cell death. Evidence suggests that cell differentiation may also be a consequence of the DDR. During the formation of the infective hypha in the phytopathogenic fungus Ustilago maydis, two DDR kinases, Atr1 and Chk1, are required to induce a G2 cell cycle arrest, which in turn is essential to display the virulence program. However, the triggering factor of DDR in this process has remained elusive. In this report we provide data suggesting that no DNA damage is associated with the activation of the DDR during the formation of the infective filament in U. maydis. We have analyzed bulk DNA replication during the formation of the infective filament, and we found no signs of impaired DNA replication. Furthermore, using RPA-GFP fusion as a surrogate marker of the presence of DNA damage, we were unable to detect any sign of DNA damage at the cellular level. In addition, neither MRN nor 9-1-1 complexes, both instrumental to transmit the DNA damage signal, are required for the induction of the above mentioned cell cycle arrest, as well as for virulence. In contrast, we have found that the claspin-like protein Mrc1, which in other systems serves as scaffold for Atr1 and Chk1, was required for both processes. We discuss possible alternative ways to trigger the DDR, independent of DNA damage, in U. maydis during virulence program activation.

  16. Conserved and Distinct Functions of the “Stunted” (StuA)-Homolog Ust1 During Cell Differentiation in the Corn Smut Fungus Ustilago maydis.

    Science.gov (United States)

    Baeza-Montañez, Lourdes; Gold, Scott E; Espeso, Eduardo A; García-Pedrajas, María D

    2015-01-01

    Ustilago maydis, causal agent of corn smut, can proliferate saprobically in a yeast form but its infectious filamentous form is an obligate parasite. Previously, we showed that Ust1, the first APSES (Asm1p, Phd1p, Sok2p, Efg1p, and StuAp) transcription factor functionally characterized in the phylum Basidiomycota, controlled morphogenesis and virulence in this species. Here, we further analyzed Ust1 function using multiple experimental approaches and determined that i) Ust1 activity was able to partially reverse stuA− conidiophore defects in Aspergillus nidulans; ii) in U. maydis, normal development and virulence were strongly dependent on precise induction or repression of Ust1 activity; iii) consistent with its role as a transcription factor regulating multiple processes, Ust1 accumulated in the nucleus at various stages of the life cycle; iv) however, it was undetectable at specific stages of pathogenic growth, indicating that Ust1 repression is part of normal development in planta; v) StuA response elements upstream of the ust1 open reading frame exhibited affinity for U. maydis DNA-binding proteins; vi) however, loss of regulated ust1 transcription had minor phenotypic effects; and vii) Ust1 was subject to post-translational phosphorylation but is not a target of cAMP signaling. Thus, the broad functional conservation between Ust1 and Ascomycota APSES proteins does not extend to the mechanisms regulating their activity.

  17. Brh2 and Rad51 promote telomere maintenance in Ustilago maydis, a new model system of DNA repair proteins at telomeres.

    Science.gov (United States)

    Yu, Eun Young; Kojic, Milorad; Holloman, William K; Lue, Neal F

    2013-07-01

    Recent studies implicate a number of DNA repair proteins in mammalian telomere maintenance. However, because several key repair proteins in mammals are missing from the well-studied budding and fission yeast, their roles at telomeres cannot be modeled in standard fungi. In this report, we explored the dimorphic fungus Ustilago maydis as an alternative model for telomere research. This fungus, which belongs to the phylum Basidiomycota, has a telomere repeat unit that is identical to the mammalian repeat, as well as a constellation of DNA repair proteins that more closely mimic the mammalian collection. We showed that the two core components of homology-directed repair (HDR) in U. maydis, namely Brh2 and Rad51, both promote telomere maintenance in telomerase positive cells, just like in mammals. In addition, we found that Brh2 is localized to telomeres in vivo, suggesting that it acts directly at chromosome ends. We surveyed a series of mutants with DNA repair defects, and found many of them to have short telomeres. Our results indicate that factors involved in DNA repair are probably also needed for optimal telomere maintenance in U. maydis, and that this fungus is a useful alternative model system for telomere research.

  18. The WOPR Protein Ros1 Is a Master Regulator of Sporogenesis and Late Effector Gene Expression in the Maize Pathogen Ustilago maydis

    Science.gov (United States)

    Tollot, Marie; Assmann, Daniela; Becker, Christian; Altmüller, Janine; Dutheil, Julien Y.; Wegner, Carl-Eric; Kahmann, Regine

    2016-01-01

    The biotrophic basidiomycete fungus Ustilago maydis causes smut disease in maize. Hallmarks of the disease are large tumors that develop on all aerial parts of the host in which dark pigmented teliospores are formed. We have identified a member of the WOPR family of transcription factors, Ros1, as major regulator of spore formation in U. maydis. ros1 expression is induced only late during infection and hence Ros1 is neither involved in plant colonization of dikaryotic fungal hyphae nor in plant tumor formation. However, during late stages of infection Ros1 is essential for fungal karyogamy, massive proliferation of diploid fungal cells and spore formation. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP-seq analyses uncovered that Ros1 remodels expression of about 30% of all U. maydis genes with 40% of these being direct targets. In total the expression of 80 transcription factor genes is controlled by Ros1. Four of the upregulated transcription factor genes were deleted and two of the mutants were affected in spore development. A large number of b-dependent genes were differentially regulated by Ros1, suggesting substantial changes in this regulatory cascade that controls filamentation and pathogenic development. Interestingly, 128 genes encoding secreted effectors involved in the establishment of biotrophic development were downregulated by Ros1 while a set of 70 “late effectors” was upregulated. These results indicate that Ros1 is a master regulator of late development in U. maydis and show that the biotrophic interaction during sporogenesis involves a drastic shift in expression of the fungal effectome including the downregulation of effectors that are essential during early stages of infection. PMID:27332891

  19. The WOPR Protein Ros1 Is a Master Regulator of Sporogenesis and Late Effector Gene Expression in the Maize Pathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Marie Tollot

    2016-06-01

    Full Text Available The biotrophic basidiomycete fungus Ustilago maydis causes smut disease in maize. Hallmarks of the disease are large tumors that develop on all aerial parts of the host in which dark pigmented teliospores are formed. We have identified a member of the WOPR family of transcription factors, Ros1, as major regulator of spore formation in U. maydis. ros1 expression is induced only late during infection and hence Ros1 is neither involved in plant colonization of dikaryotic fungal hyphae nor in plant tumor formation. However, during late stages of infection Ros1 is essential for fungal karyogamy, massive proliferation of diploid fungal cells and spore formation. Premature expression of ros1 revealed that Ros1 counteracts the b-dependent filamentation program and induces morphological alterations resembling the early steps of sporogenesis. Transcriptional profiling and ChIP-seq analyses uncovered that Ros1 remodels expression of about 30% of all U. maydis genes with 40% of these being direct targets. In total the expression of 80 transcription factor genes is controlled by Ros1. Four of the upregulated transcription factor genes were deleted and two of the mutants were affected in spore development. A large number of b-dependent genes were differentially regulated by Ros1, suggesting substantial changes in this regulatory cascade that controls filamentation and pathogenic development. Interestingly, 128 genes encoding secreted effectors involved in the establishment of biotrophic development were downregulated by Ros1 while a set of 70 "late effectors" was upregulated. These results indicate that Ros1 is a master regulator of late development in U. maydis and show that the biotrophic interaction during sporogenesis involves a drastic shift in expression of the fungal effectome including the downregulation of effectors that are essential during early stages of infection.

  20. The distinct interaction between cell cycle regulation and the widely conserved morphogenesis-related (MOR) pathway in the fungus Ustilago maydis determines morphology.

    Science.gov (United States)

    Sartorel, Elodie; Pérez-Martín, José

    2012-10-01

    The morphogenesis-related NDR kinase (MOR) pathway regulates morphogenesis in fungi. In spite of the high conservation of its components, impairing their functions results in highly divergent cellular responses depending on the fungal species. The reasons for such differences are unclear. Here we propose that the species-specific connections between cell cycle regulation and the MOR pathway could be partly responsible for these divergences. We based our conclusion on the characterization of the MOR pathway in the fungus Ustilago maydis. Each gene that encodes proteins of this pathway in U. maydis was deleted. All mutants exhibited a constitutive hyperpolarized growth, contrasting with the loss of polarity observed in other fungi. Using a conditional allele of the central NDR kinase Ukc1, we found that impairing MOR function resulted in a prolonged G2 phase. This cell cycle delay appears to be the consequence of an increase in Cdk1 inhibitory phosphorylation. Strikingly, prevention of the inhibitory Cdk1 phosphorylation abolished the hyperpolarized growth associated with MOR pathway depletion. We found that the prolonged G2 phase resulted in higher levels of expression of crk1, a conserved kinase that promotes polar growth in U. maydis. Deletion of crk1 also abolished the dramatic activation of polar growth in cells lacking the MOR pathway. Taken together, our results suggest that Cdk1 inhibitory phosphorylation may act as an integrator of signaling cascades regulating fungal morphogenesis and that the distinct morphological response observed in U. maydis upon impairment of the MOR pathway could be due to a cell cycle deregulation.

  1. Role of Hsl7 in morphology and pathogenicity and its interaction with other signaling components in the plant pathogen Ustilago maydis.

    Science.gov (United States)

    Lovely, C Ben; Aulakh, Kavita Burman; Perlin, Michael H

    2011-07-01

    The phytopathogenic fungus Ustilago maydis undergoes a dimorphic transition in response to mating pheromone, host, and environmental cues. On a solid medium deficient in ammonium (SLAD [0.17% yeast nitrogen base without ammonium sulfate or amino acids, 2% dextrose, 50 μM ammonium sulfate]), U. maydis produces a filamentous colony morphology, while in liquid SLAD, the cells do not form filaments. The p21-activated protein kinases (PAKs) play a substantial role in regulating the dimorphic transition in fungi. The PAK-like Ste20 homologue Smu1 is required for a normal response to pheromone, via upregulation of pheromone expression, and virulence, and its disruption affects both processes. Our experiments suggest that Smu1 also regulates cell length and the filamentous response on solid SLAD medium. Yeast two-hybrid analysis suggested an Hsl7 homologue as a potential interacting partner of Smu1, and a unique open reading frame for such an arginine methyltransferase was detected in the U. maydis genome sequence. Hsl7 regulates cell length and the filamentous response to solid SLAD in a fashion opposite to that of Smu1, but neither overexpression nor disruption of hsl7 attenuates virulence. Simultaneous disruption of hsl7 and overexpression of smu1 lead to a hyperfilamentous response on solid SLAD. Moreover, only this double mutant strain forms filaments in liquid SLAD. The double mutant strain was also significantly reduced in virulence. A similar filamentous response in both solid and liquid SLAD was observed in strains lacking another PAK-like protein kinase involved in cytokinesis and polar growth, Cla4. Our data suggest that Hsl7 may regulate cell cycle progression, while both Smu1 and Cla4 appear to be involved in the filamentous response in U. maydis.

  2. The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity.

    Science.gov (United States)

    Brundiek, Henrike; Saß, Stefan; Evitt, Andrew; Kourist, Robert; Bornscheuer, Uwe T

    2012-04-01

    The Ustilago maydis lipase UM03410 belongs to the mostly unexplored Candida antarctica lipase (CAL-A) subfamily. The two lipases with [corrected] the highest identity are a lipase from Sporisorium reilianum and the prototypic CAL-A. In contrast to the other CAL-A-type lipases, this hypothetical U. maydis lipase is annotated to possess a prolonged N-terminus of unknown function. Here, we show for the first time the recombinant expression of two versions of lipase UM03410: the full-length form (lipUMf) and an Nterminally truncated form (lipUMs). For comparison to the prototype, the expression of recombinant CAL-A in E. coli was investigated. Although both forms of lipase UM03410 could be expressed functionally in E. coli, the N-terminally truncated form (lipUMs) demonstrated significantly higher activities towards p-nitrophenyl esters. The functional expression of the N-terminally truncated lipase was further optimized by the appropriate choice of the E. coli strain, lowering the cultivation temperature to 20 °C and enrichment of the cultivation medium with glucose. Primary characteristics of the recombinant lipase are its pH optimum in the range of 6.5-7.0 and its temperature optimum at 55 °C. As is typical for lipases, lipUM03410 shows preference for long chain fatty acid esters with myristic acid ester (C14:0 ester) being the most preferred one.More importantly, lipUMs exhibits an inherent preference for C18:1Δ9 trans and C18:1Δ11 trans-fatty acid esters similar to CAL-A. Therefore, the short form of this U. maydis lipase is the only other currently known lipase with a distinct trans-fatty acid selectivity.

  3. Investigating the Ustilago maydis/Zea mays pathosystem: transcriptional responses and novel functional aspects of a fungal calcineurin regulatory B subunit.

    Science.gov (United States)

    Donaldson, Michael E; Meng, Shaowu; Gagarinova, Alla; Babu, Mohan; Lambie, Scott C; Swiadek, Alexander A; Saville, Barry J

    2013-01-01

    The sustainable control of basidiomycete biotrophic plant pathogenesis requires an understanding of host responses to infection, as well as the identification and functional analysis of fungal genes involved in disease development. The creation and analysis of a suppressive subtractive hybridization (SSH) cDNA library from Ustilago maydis-infected Zea mays seedlings enabled the identification of fungal and plant genes expressed during disease development, and uncovered new insights into the interactions of this model system. Candidate U. maydis pathogenesis genes were identified by using the current SSH cDNA library analysis, and by knowledge generated from previous cDNA microarray and comparative genomic analyses. These identifications were supported by the independent determination of transcript level changes in different cell-types and during pathogenic development. The basidiomycete specific um01632, the highly in planta expressed um03046 (zig1), and the calcineurin regulatory B subunit (um10226, cnb1), were chosen for deletion experiments. um01632 and zig1 mutants showed no difference in morphology and did not have a statistically significant impact on pathogenesis. cnb1 mutants had a distinct cell division phenotype and reduced virulence in seedling assays. Infections with reciprocal wild-type×Δcnb1 haploid strain crosses revealed that the wild-type allele was unable to fully compensate for the lack of a second cnb1 allele. This haploinsufficiency was undetected in other fungal cnb1 mutational analyses. The reported data improves U. maydis genome annotation and expands on the current understanding of pathogenesis genes in this model basidiomycete.

  4. MRN- and 9-1-1-Independent Activation of the ATR-Chk1 Pathway during the Induction of the Virulence Program in the Phytopathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    María Tenorio-Gómez

    Full Text Available DNA damage response (DDR leads to DNA repair, and depending on the extent of the damage, to further events, including cell death. Evidence suggests that cell differentiation may also be a consequence of the DDR. During the formation of the infective hypha in the phytopathogenic fungus Ustilago maydis, two DDR kinases, Atr1 and Chk1, are required to induce a G2 cell cycle arrest, which in turn is essential to display the virulence program. However, the triggering factor of DDR in this process has remained elusive. In this report we provide data suggesting that no DNA damage is associated with the activation of the DDR during the formation of the infective filament in U. maydis. We have analyzed bulk DNA replication during the formation of the infective filament, and we found no signs of impaired DNA replication. Furthermore, using RPA-GFP fusion as a surrogate marker of the presence of DNA damage, we were unable to detect any sign of DNA damage at the cellular level. In addition, neither MRN nor 9-1-1 complexes, both instrumental to transmit the DNA damage signal, are required for the induction of the above mentioned cell cycle arrest, as well as for virulence. In contrast, we have found that the claspin-like protein Mrc1, which in other systems serves as scaffold for Atr1 and Chk1, was required for both processes. We discuss possible alternative ways to trigger the DDR, independent of DNA damage, in U. maydis during virulence program activation.

  5. The mitochondrial LSU rRNA group II intron of Ustilago maydis encodes an active homing endonuclease likely involved in intron mobility.

    Directory of Open Access Journals (Sweden)

    Anja Pfeifer

    Full Text Available BACKGROUND: The a2 mating type locus gene lga2 is critical for uniparental mitochondrial DNA inheritance during sexual development of Ustilago maydis. Specifically, the absence of lga2 results in biparental inheritance, along with efficient transfer of intronic regions in the large subunit rRNA gene between parental molecules. However, the underlying role of the predicted LAGLIDADG homing endonuclease gene I-UmaI located within the group II intron LRII1 has remained unresolved. METHODOLOGY/PRINCIPAL FINDINGS: We have investigated the enzymatic activity of I-UmaI in vitro based on expression of a tagged full-length and a naturally occurring mutant derivative, which harbors only the N-terminal LAGLIDADG domain. This confirmed Mg²⁺-dependent endonuclease activity and cleavage at the LRII1 insertion site to generate four base pair extensions with 3' overhangs. Specifically, I-UmaI recognizes an asymmetric DNA sequence with a minimum length of 14 base pairs (5'-GACGGGAAGACCCT-3' and tolerates subtle base pair substitutions within the homing site. Enzymatic analysis of the mutant variant indicated a correlation between the activity in vitro and intron homing. Bioinformatic analyses revealed that putatively functional or former functional I-UmaI homologs are confined to a few members within the Ustilaginales and Agaricales, including the phylogenetically distant species Lentinula edodes, and are linked to group II introns inserted into homologous positions in the LSU rDNA. CONCLUSIONS/SIGNIFICANCE: The present data provide strong evidence that intron homing efficiently operates under conditions of biparental inheritance in U. maydis. Conversely, uniparental inheritance may be critical to restrict the transmission of mobile introns. Bioinformatic analyses suggest that I-UmaI-associated introns have been acquired independently in distant taxa and are more widespread than anticipated from available genomic data.

  6. Transcriptomic analysis of the role of Rim101/PacC in the adaptation of Ustilago maydis to an alkaline environment.

    Science.gov (United States)

    Franco-Frías, Eduardo; Ruiz-Herrera, Jose; Aréchiga-Carvajal, Elva T

    2014-09-01

    Alkaline pH triggers an adaptation mechanism in fungi that is mediated by Rim101/PacCp, a zinc finger transcription factor. To identify the genes under its control in Ustilago maydis, we performed microarray analyses, comparing gene expression in a wild-type strain versus a rim101/pacC mutation strain of the fungus. In this study we obtained evidence of the large number of genes regulated mostly directly, but also indirectly (probably through regulation of other transcription factors), by Rim101/PacCp, including proteins involved in a large number of physiological activities of the fungus. Our analyses suggest that the response to alkaline conditions under the control of the Pal/Rim pathway involves changes in the cell wall and plasma membrane through alterations in their lipid, protein and polysaccharide composition, changes in cell polarity, actin cytoskeleton organization, and budding patterns. Also as expected, adaptation involves regulation by Rim101/PacC of genes involved in meiotic functions, such as recombination and segregation, and expression of genes involved in ion and nutrient transport, as well as general vacuole functions.

  7. Septation of infectious hyphae is critical for appressoria formation and virulence in the smut fungus Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Johannes Freitag

    2011-05-01

    Full Text Available Differentiation of hyphae into specialized infection structures, known as appressoria, is a common feature of plant pathogenic fungi that penetrate the plant cuticle. Appressorium formation in U. maydis is triggered by environmental signals but the molecular mechanism of this hyphal differentiation is largely unknown. Infectious hyphae grow on the leaf surface by inserting regularly spaced retraction septa at the distal end of the tip cell leaving empty sections of collapsed hyphae behind. Here we show that formation of retraction septa is critical for appressorium formation and virulence in U. maydis. We demonstrate that the diaphanous-related formin Drf1 is necessary for actomyosin ring formation during septation of infectious hyphae. Drf1 acts as an effector of a Cdc42 GTPase signaling module, which also consists of the Cdc42-specific guanine nucleotide exchange factor Don1 and the Ste20-like kinase Don3. Deletion of drf1, don1 or don3 abolished formation of retraction septa resulting in reduced virulence. Appressorium formation in these mutants was not completely blocked but infection structures were found only at the tip of short filaments indicating that retraction septa are necessary for appressorium formation in extended infectious hyphae. In addition, appressoria of drf1 mutants penetrated the plant tissue less frequently.

  8. Mutation avoidance and DNA repair proficiency in Ustilago maydis are differentially lost with progressive truncation of the REC1 gene product

    Energy Technology Data Exchange (ETDEWEB)

    Onel, K.; Thelen, M.P.; Ferguson, D.O.; Bennett, R.L.; Holloman, W.K. [Cornell Univ. Medical College, NY, NY (United States)

    1995-10-01

    The REC1 gene of Ustilago maydis has an uninterrupted open reading frame, predicted from the genomic sequence to encode a protein of 522 amino acid residues. Nevertheless, an intron is present, and functional activity of the gene in mitotic cells requires an RNA processing event to remove the intron. This results in a change in reading frame and production of a protein of 463 amino acid residues. The 3{prime}{r_arrow}5{prime} exonuclease activity of proteins derived form the REC1 genomic open reading frame, the intronless open reading frame, and several mutants was investigated. The mutants included a series of deletions constructed by removing restriction fragments at the 3{prime} end of the cloned REC1 gene and a set of mutant alleles previously isolated in screens for radiation sensitivity. The results indicated that elimination of the C-terminal third of the protein did not result in a serious reduction in 3{prime}{r_arrow}5{prime} exonuclease activity, but deletion into the midsection caused a severe loss of activity. The biological activity of the rec1-1 allele, which encodes a truncated polypeptide with full 3{prime}{r_arrow}5{prime} exonuclease activity, and the rec1-5 allele, which encodes a more severely truncated polypeptide with no exonuclease activity, was investigated. The two mutants were equally sensitive to the lethal effect of UV light, but the spontaneous mutation rate was elevated 10-fold over the wild-type rate in the rec1-1 mutant and 100-fold in the rec1-5 mutant. The elevated spontaneous mutation rate correlated with the ablation of exonuclease activity, but the radiation sensitivity did not. These results indicate that the C-terminal portion of the Rec1 protein is not essential for exonuclease activity but is crucial in the role of REC1 in DNA damage repair. 49 refs., 3 figs., 1 tab.

  9. 玉米瘤黑粉菌的遗传交配型%The Mating Type of Ustilago maydis

    Institute of Scientific and Technical Information of China (English)

    陈三凤; 刘德虎; 李季伦

    2000-01-01

    @@玉米瘤黑粉病是玉米的一种重要病害,普遍分布于世界各玉米产区,我国各地也有不同程度的发生,主要症状是在玉米的茎、叶、雄花、雌穗等部位形成肿瘤[1].其病原菌为玉米瘤黑粉菌(Ustilalgo maydis),属于担子菌亚门,异宗配合.在玉米瘤黑粉菌的生活史中,有两种不同形态的细胞,即单倍体细胞(担孢子)和双核菌丝体.单倍体细胞没有致病性,在特定培养基上芽殖产生"酵母”状菌落.不同遗传型的单倍体细胞融合形成双核菌丝,双核菌丝能在寄主植物体内迅速发育,刺激寄主组织形成肿瘤,并继而经过细胞核融合,产生双倍体的冬孢子[2].

  10. 玉米黑粉菌实时荧光定量PCR检测方法的建立%Development of Real-Time Quantitative PCR Method for Detection of Ustilago maydis

    Institute of Scientific and Technical Information of China (English)

    魏国荣; 李凯瑞; 黄雪玲

    2014-01-01

    目的:建立利用实时荧光定量PCR对玉米黑粉菌[Ustilago maydis(DC.) Corda]快速定量检测的方法.方法:以玉米黑粉菌β actin基因为内参基因,以含有βactin基因的具浓度梯度pMD19 T-Simple质粒为标准品,接种黑粉菌后不同时间点的玉米叶片作为样本用以检验该方法的实用性.结果:建立了利用实时荧光定量PCR对玉米黑粉菌进行定量分析的方法.该方法重复性好、特异性强、灵敏性高,对玉米黑粉菌的最低检出率为19拷贝/反应,可以准确检测经注射接种后1d的玉米叶片中的黑粉菌量.结论:建立的实时荧光定量PCR方法可用于玉米黑粉菌菌量的检测.

  11. 玉米黑粉菌培养条件响应面法优化研究%Optimization of cultivation conditions for extracellular polysaccharide fermented by Ustilago maydis using response surface methodology

    Institute of Scientific and Technical Information of China (English)

    王永斌; 王允祥

    2006-01-01

    采用P1ackett-Burman设计(Plackett-Burman Design,P-B)对影响玉米黑粉菌(Ustilago maydis)AS 5.128发酵胞外多糖的内在和外在因素进行了筛选,所选取的6个相关因素为:初始pH、培养温度、发酵时间、装液量、接种量、摇床转速.在此基础上,采用响应曲面法(Response Surface Methodology,RSM)对影响玉米黑粉菌发酵胞外多糖的关键影响因素培养温度、发酵时间和接种量的最佳水平范围作了进一步的研究,通过对二次多项回归方程求解得知,培养温度2637℃、发酵时间7.03d和接种量9.49%时,胞外多糖产量的最大预测值为1650.92μg/mL.

  12. Draft Genome Sequence of Ustilago trichophora RK089, a Promising Malic Acid Producer

    Science.gov (United States)

    Zambanini, Thiemo; Buescher, Joerg M.; Meurer, Guido; Blank, Lars M.

    2016-01-01

    The basidiomycetous smut fungus Ustilago trichophora RK089 produces malate from glycerol. De novo genome sequencing revealed a 20.7-Mbp genome (301 gap-closed contigs, 246 scaffolds). A comparison to the genome of Ustilago maydis 521 revealed all essential genes for malate production from glycerol contributing to metabolic engineering for improving malate production. PMID:27469969

  13. Estudo dos mecanismos de distribuição e motilidade peroxissomal baseados em microtubulos, utilizando o fungo filamentoso U. maydis

    OpenAIRE

    Guimarães, Ana Sofia da Cunha

    2016-01-01

    Peroxisomes are ubiquitous subcellular organelles, which fulfil important metabolic functions, notably the β-oxidation of fatty acids and the metabolism of hydrogen peroxide, and are thus essential for human health and development. The filamentous fungus Ustilago maydis is a biotrophic, basidiomycete responsible for corn smut disease. U. maydis exhibits several features similar to mammals including polar growth, microtubule-dependent organelle trafficking and open mitosis. I...

  14. 玉米黑粉菌CYP51稀有密码子和mRNA二级结构分析及与杀菌剂戊唑醇分子对接%0Analysis of Rare Codon and mRNA Structure About Ustilago maydis CYP51 and Molecular Docking With Fungicide Tebuconazole

    Institute of Scientific and Technical Information of China (English)

    李书祥; 韩睿; 袁利玲; 熊丽; 袁永泽; 杨江科; 闫云君; 刘德立

    2011-01-01

    通过截短玉米黑粉菌CYP51(P450-14DM,UmCYP51)基因(去除编码跨膜区部分)和选取不同的表达载体,构建了9种重组表达质粒,在大肠杆菌中进行UmCYP51基因的表达,发现只有BL21(DE3)/PET32-Um-35重组表达工程菌获得了表达.对稀有密码子和mRNA翻译起始区二级结构进行分析,结果表明稀有密码子和mRNA翻译起始区二级结构对UmCYP51蛋白的表达都有影响.适用于稀有密码子表达的菌株Rosetta(DE3)不利于UmCYP51蛋白的表达;同时只有翻译起始区二级结构自由能值最低的重组载体pET32-Um-35可以表达.为了设计以UmCYP51为靶标的新型抗真菌抑制剂,基于最新解析的真核生物人类的CYP51晶体结构,利用同源模建的方法构建了UmCYP51的三维结构并进行了分子动力学模拟优化.通过与商品化杀菌剂戊唑醇进行分子对接获得了此类抑制剂与UmCYP51的理论结合方式,阐述了戊唑醇分子的杀菌机理,为开发新型的抗真菌抑制剂奠定了基础.%To get a better optimization expression of the Ustilago maydis CYP51 (P450-14DM, UmCYP51) protein in E. Coli BL21 (DE3), the different lengths of UmCYP51 gene that lacked the coding region for the putative membrane-spanning segment of the N-terminus were truncated. The first one is the wild type, the second one with 20 amino acids (60 base pairs) in N-terminus was truncated and the third one with 35 amino acids (105 base pairs) was truncated. Then these genes were incorporated into different expression vectors (pET28, pET32 and pGEX-KG) to construct nine recombinant expression plasmids (pET28-Um, pET28-Um-20, pET28-Um-35, pET32-Um, pET32-Um-20, pET32-Um-35, pGEXKG-Um, pGEXKG-Um-20 and pGEXKG-Um-35). The expression of recombinant plasmids were performed using 0.5 mmol/L of isopropyl β-D-thiogalactoside (IPTG) at 30°C. The culture harvested every 2 h up to 8 h. It was found that only recombinant plasmid pET32-Um-35 was expressed in E. Coli BL21(DE3). Codon

  15. Helminthosporium Maydis en Colombia

    Directory of Open Access Journals (Sweden)

    Castaño A. J. J.

    1971-09-01

    Full Text Available Antes de Diciembre de 1970, había dudas acerca de la ocurrencia de Helminthosporium maydis Nisikado & Miyake, en los cultivos demaíz en Colombia. Es bien sabido que este hongo ha causado, a través de 1970-1971, desastres en extensos cultivos de maíz de la zona maicera de los Estados Unidos, y que en aquel país las entidades federales y estatales se han visto precisadas a dedicar grandes esfuerzos técnicos y científicos, como también apreciables recursos económicos, para solucionar oportunamente este grave problema. De una muestra de hojas de maíz procedente del Centro Nacional de Investigaciones Agropecuarias "Turipaná", recibida por el suscrito como consulta fitosanitaria, en Diciembre de 1970, fue aislado un hongo con características que correspondían a las del Helminthosporium maydis Nisikado & Miyake. Para más evidencia en cuanto a la identificación de la especie, un cultivo puro del hongo fue enviado con tal fin al doctor Lekh R. Batra, micólogo del Departamento de Agricultura de los Estados Unidos, quien a su vez también remitió un duplicado al doctor E. S. Luttrell, profesor del Departamento de Fitopatología y Genética de Plantas de la Universidad de Georgia, para su estudio y determinación. El interés Que tan gentilme'.1.te dedicaron dichos científicos al estudio específico del hongo, dio como resultado su plena identificación como H. maydis, locual reconfirmó también la nuestra, y por consiguiente se corroboró de manera indudable la ocurrencia del hongo como fitoparásito del maíz en Colombia. En este caso, el paso siguiente podría ser el de determinar a cuál raza corresponde el hongo, si a la universal, o sea a la Raza O, o a la Raza T. Esta última es precisamente la que en los Estados Unidos ha presentado alarmantes proporciones epifitóticas durante 1970-1971, especialmente en los maíces cuyo citoplasma macho estéril posee factores provenientes de los maíces de Texas (Tms.

  16. KP4 to control Ustilago tritici in wheat: Enhanced greenhouse resistance to loose smut and changes in transcript abundance of pathogen related genes in infected KP4 plants.

    Science.gov (United States)

    Quijano, Carolina Diaz; Wichmann, Fabienne; Schlaich, Thomas; Fammartino, Alessandro; Huckauf, Jana; Schmidt, Kerstin; Unger, Christoph; Broer, Inge; Sautter, Christof

    2016-09-01

    Ustilago tritici causes loose smut, which is a seed-borne fungal disease of wheat, and responsible for yield losses up to 40%. Loose smut is a threat to seed production in developing countries where small scale farmers use their own harvest as seed material. The killer protein 4 (KP4) is a virally encoded toxin from Ustilago maydis and inhibits growth of susceptible races of fungi from the Ustilaginales. Enhanced resistance in KP4 wheat to stinking smut, which is caused by Tilletia caries, had been reported earlier. We show that KP4 in genetically engineered wheat increased resistance to loose smut up to 60% compared to the non-KP4 control under greenhouse conditions. This enhanced resistance is dose and race dependent. The overexpression of the transgene kp4 and its effect on fungal growth have indirect effects on the expression of endogenous pathogen defense genes.

  17. KP4 to control Ustilago tritici in wheat: Enhanced greenhouse resistance to loose smut and changes in transcript abundance of pathogen related genes in infected KP4 plants

    Directory of Open Access Journals (Sweden)

    Carolina Diaz Quijano

    2016-09-01

    Full Text Available Ustilago tritici causes loose smut, which is a seed-borne fungal disease of wheat, and responsible for yield losses up to 40%. Loose smut is a threat to seed production in developing countries where small scale farmers use their own harvest as seed material. The killer protein 4 (KP4 is a virally encoded toxin from Ustilago maydis and inhibits growth of susceptible races of fungi from the Ustilaginales. Enhanced resistance in KP4 wheat to stinking smut, which is caused by Tilletia caries, had been reported earlier. We show that KP4 in genetically engineered wheat increased resistance to loose smut up to 60% compared to the non-KP4 control under greenhouse conditions. This enhanced resistance is dose and race dependent. The overexpression of the transgene kp4 and its effect on fungal growth have indirect effects on the expression of endogenous pathogen defense genes.

  18. THE IDENTITY OF USTILAGO AMADELPHA VAR. GLABRIUSCULA

    Directory of Open Access Journals (Sweden)

    MIEN A. RIFAI

    1980-03-01

    Full Text Available The host of this smut variety is identified as Panicum repens L. Thefungus is considered conspecific with Ustilago overeemii Cif., which sub-sequently is reclassified as Sporisorium overeemii (Cif. Rifai, comb. nov.

  19. Compensatory role for Rad52 during recombinational repair in Ustilago maydis

    DEFF Research Database (Denmark)

    Kojic, Milorad; Mao, Ninghui; Zhou, Qingwen

    2008-01-01

    in spontaneous mutator activity, allelic recombination or meiosis. GFP-Rad51 foci were formed in rad52 cells following DNA damage, but were initially less intense than normal suggesting a possible role for Rad52 in formation of the Rad51 nucleoprotein filament. A search for interacting genes that confer...

  20. Ultrastructure and phylogeny of Ustilago coicis *

    Science.gov (United States)

    Zhang, Jing-ze; Guan, Pei-gang; Tao, Gang; Ojaghian, Mohammad Reza; Hyde, Kevin David

    2013-01-01

    Ustilago coicis causes serious smut on Coix lacryma-jobi in Dayang Town, Jinyun County, Zhejiang Province of China. In this paper, ultrastructural assessments on fungus-host interactions and teliospore development are presented, and molecular phylogenetic analyses have been done to elucidate the phylogenetic placement of the taxon. Hyphal growth within infected tissues was both intracellular and intercellular and on the surface of fungus-host interaction, and the fungal cell wall and the invaginated host plasma membrane were separated by a sheath comprising two distinct layers between the fungal cell wall and the invaginated host plasma membrane. Ornamentation development of teliospore walls was unique as they appeared to be originated from the exosporium. In addition, internal transcribed spacer (ITS) and large subunit (LSU) sequence data showed that U. coicis is closely related to Ustilago trichophora which infects grass species of the genus Echinochloa (Poaceae). PMID:23549851

  1. Ultrastructure and phylogeny of Ustilago coicis

    Institute of Scientific and Technical Information of China (English)

    Jing-ze ZHANG; Pei-gang GUAN; Gang TAO; Mohammad Reza OJAGHIAN; Kevin David HYDE

    2013-01-01

    Ustilago coicis causes serious smut on Coix lacryma-jobi in Dayang Town,Jinyun County,Zhejiang Province of China.In this paper,ultrastructural assessments on fungus-host interactions and teliospore development are presented,and molecular phylogenetic analyses have been done to elucidate the phylogenetic placement of the taxon.Hyphal growth within infected tissues was both intracellular and intercellular and on the surface of fungus-host interaction,and the fungal cell wall and the invaginated host plasma membrane were separated by a sheath comprising two distinct layers between the fungal cell wall and the invaginated host plasma membrane.Ornamentation development of teliospore walls was unique as they appeared to be originated from the exosporium.In addition,internal transcribed spacer (ITS) and large subunit (LSU) sequence data showed that U.coicis is closely related to Ustilago trichophora which infects grass species of the genus Echinochloa (Poaceae).

  2. 玉米黑粉菌孢子粉的无机元素和氨基酸含量测定%Determination of Inorganic Elements and Amino Acids from Spore Powder of Usitilago maydis

    Institute of Scientific and Technical Information of China (English)

    王辉; 斯琴格日乐; 马端辉; 包海鹰

    2006-01-01

    对玉米黑粉菌(Ustilago maydis DC.Corola.)孢子粉的无机元素和氨基酸含量进行了测定.结果表明:玉米黑粉菌孢子粉含有丰富的无机元素,其中铁、钙、镁、磷、锌、锰含量分别为100μg/g、3 561 μg/g、1 382μg/g、3 500μg/g、101μg/g和24μg/g;同时含有8种必须由食物蛋白质提供的氨基酸,以苏氨酸含量最多,为340.18 mg/100g.

  3. Interactions between Fusarium verticillioides, Ustilago maydis, and Zea mays: an endophyte, a pathogen, and their shared plant host

    Science.gov (United States)

    Highly diverse communities of microbial symbionts occupy eukaryotic organisms, including plants. While many well-studied symbionts may be characterized as either parasites or as mutualists, the prevalent but cryptic endophytic fungi are less easily qualified because they do not cause observable symp...

  4. An immunity-triggering effector from the Barley smut fungus Ustilago hordei resides in an Ustilaginaceae-specific cluster bearing signs of transposable element-assisted evolution.

    Science.gov (United States)

    Ali, Shawkat; Laurie, John D; Linning, Rob; Cervantes-Chávez, José Antonio; Gaudet, Denis; Bakkeren, Guus

    2014-07-01

    The basidiomycete smut fungus Ustilago hordei was previously shown to comprise isolates that are avirulent on various barley host cultivars. Through genetic crosses we had revealed that a dominant avirulence locus UhAvr1 which triggers immunity in barley cultivar Hannchen harboring resistance gene Ruh1, resided within an 80-kb region. DNA sequence analysis of this genetically delimited region uncovered the presence of 7 candidate secreted effector proteins. Sequence comparison of their coding sequences among virulent and avirulent parental and field isolates could not distinguish UhAvr1 candidates. Systematic deletion and complementation analyses revealed that UhAvr1 is UHOR_10022 which codes for a small effector protein of 171 amino acids with a predicted 19 amino acid signal peptide. Virulence in the parental isolate is caused by the insertion of a fragment of 5.5 kb with similarity to a common U. hordei transposable element (TE), interrupting the promoter of UhAvr1 and thereby changing expression and hence recognition of UhAVR1p. This rearrangement is likely caused by activities of TEs and variation is seen among isolates. Using GFP-chimeric constructs we show that UhAvr1 is induced only in mated dikaryotic hyphae upon sensing and infecting barley coleoptile cells. When infecting Hannchen, UhAVR1p causes local callose deposition and the production of reactive oxygen species and necrosis indicative of the immune response. UhAvr1 does not contribute significantly to overall virulence. UhAvr1 is located in a cluster of ten effectors with several paralogs and over 50% of TEs. This cluster is syntenous with clusters in closely-related U. maydis and Sporisorium reilianum. In these corn-infecting species, these clusters harbor however more and further diversified homologous effector families but very few TEs. This increased variability may have resulted from past selection pressure by resistance genes since U. maydis is not known to trigger immunity in its corn host.

  5. Amplified Fragment Length Polymorphism Diversity in Cephalosporium maydis from Egypt.

    Science.gov (United States)

    Saleh, Amgad A; Zeller, Kurt A; Ismael, Abou-Serie M; Fahmy, Zeinab M; El-Assiuty, Elhamy M; Leslie, John F

    2003-07-01

    ABSTRACT Cephalosporium maydis, the causal agent of late wilt of maize, was first described in Egypt in the 1960s, where it can cause yield losses of up to 40% in susceptible plantings. We characterized 866 isolates of C. maydis collected from 14 governates in Egypt, 7 in the Nile River Delta and 7 in southern (Middle and Upper) Egypt, with amplified fragment length polymorphism (AFLP) markers. The four AFLP primer-pair combinations generated 68 bands, 25 of which were polymorphic, resulting in 52 clonal haplotypes that clustered the 866 isolates into four phylogenetic lineages. Three lineages were found in both the Nile River Delta and southern Egypt. Lineage IV, the most diverse group (20 haplotypes), was recovered only from governates in the Nile River Delta. In some locations, one lineage dominated (up to 98% of the isolates recovered) and, from some fields, only a single haplotype was recovered. Under field conditions in Egypt, there is no evidence that C. maydis reproduces sexually. The nonuniform geographic distribution of the pathogen lineages within the country could be due to differences in climate or in the farming system, because host material differs in susceptibility and C. maydis lineages differ in pathogenicity.

  6. Transmission of tenocarpella maydis by maize seeds

    Directory of Open Access Journals (Sweden)

    Carolina da Silva Siqueira

    2016-06-01

    Full Text Available ABSTRACT tenocarpella maydis is one of the main fungi associated with maize seeds, being a causative agent of stalk and ear rot, a disease which causes considerable losses for crop-producing regions in Brazil. The organism is considered to be a pest, subject to sanitary standardisation in current programs of seed certification in the country. The aim of this study was to evaluate the transmission rate of the fungus from infected maize seeds. Seeds were inoculated with two isolates using a method of physiological conditioning, in which the seeds are kept in contact with colonies of the fungus for 24 (P1, 48 (P2, 72 (P3 and 96 (P4 hours. Two cultivars were used, one susceptible (C1 and one moderately resistant (C2, and the trial carried out at two temperatures (20 ºC and 25 ºC. The inoculated seeds were distributed individually into plastic cups containing substrate. The plants were evaluated daily for stand and the appearance of post-emergent symptoms. Based on the number of dead seeds, transmission rates reached a maximum of 90.5% at the P4 inoculum potential, this rate being greater than transmission rates achieved for symptomatic and asymptomatic infection in emerged plants. For the total transmission rate, transmission of the pathogen was seen at all inoculum potentials; these values varying from 25% for cultivar C2 at potential P1 and a temperature of 20 ºC, to 93% for cultivar C2 at potential P3 and a temperature of 25 ºC.

  7. Cephalosporium maydis is a distinct species in the Gaeumannomyces-Harpophora species complex.

    Science.gov (United States)

    Saleh, Amgad A; Leslie, John F

    2004-01-01

    Cephalosporium maydis is an important plant pathogen whose phylogenetic position relative to other fungi has not been established clearly. We compared strains of C. maydis, strains from several other plant-pathogenic Cephalosporium spp. and several possible relatives within the Gaeumannomyces-Harpophora species complex, to which C. maydis has been suggested to belong based on previous preliminary DNA sequence analyses. DNA sequences of the nuclear genes encoding the rDNA ITS region, β-tubulin, histone H3, and MAT-2 support the hypothesis that C. maydis is a distinct taxon within the Gaeumannomyces-Harpophora species complex. Based on amplified fragment length polymorphism (AFLP) profiles, C. maydis also is distinct from the other tested species of Cephalosporium, Phialophora sensu lato and members of Gaeumannomyces-Harpophora species complex, which supports its classification as Harpophora maydis. Oligonucleotide primers for H. maydis were developed that can be used in a PCR diagnostic protocol to rapidly and reliably detect and identify this pathogen. These diagnostic PCR primers will aid the detection of H. maydis in diseased maize because this fungus can be difficult to detect and isolate, and the movement of authentic cultures may be limited by quarantine restrictions.

  8. An immunity-triggering effector from the Barley smut fungus Ustilago hordei resides in an Ustilaginaceae-specific cluster bearing signs of transposable element-assisted evolution.

    Directory of Open Access Journals (Sweden)

    Shawkat Ali

    2014-07-01

    Full Text Available The basidiomycete smut fungus Ustilago hordei was previously shown to comprise isolates that are avirulent on various barley host cultivars. Through genetic crosses we had revealed that a dominant avirulence locus UhAvr1 which triggers immunity in barley cultivar Hannchen harboring resistance gene Ruh1, resided within an 80-kb region. DNA sequence analysis of this genetically delimited region uncovered the presence of 7 candidate secreted effector proteins. Sequence comparison of their coding sequences among virulent and avirulent parental and field isolates could not distinguish UhAvr1 candidates. Systematic deletion and complementation analyses revealed that UhAvr1 is UHOR_10022 which codes for a small effector protein of 171 amino acids with a predicted 19 amino acid signal peptide. Virulence in the parental isolate is caused by the insertion of a fragment of 5.5 kb with similarity to a common U. hordei transposable element (TE, interrupting the promoter of UhAvr1 and thereby changing expression and hence recognition of UhAVR1p. This rearrangement is likely caused by activities of TEs and variation is seen among isolates. Using GFP-chimeric constructs we show that UhAvr1 is induced only in mated dikaryotic hyphae upon sensing and infecting barley coleoptile cells. When infecting Hannchen, UhAVR1p causes local callose deposition and the production of reactive oxygen species and necrosis indicative of the immune response. UhAvr1 does not contribute significantly to overall virulence. UhAvr1 is located in a cluster of ten effectors with several paralogs and over 50% of TEs. This cluster is syntenous with clusters in closely-related U. maydis and Sporisorium reilianum. In these corn-infecting species, these clusters harbor however more and further diversified homologous effector families but very few TEs. This increased variability may have resulted from past selection pressure by resistance genes since U. maydis is not known to trigger immunity

  9. An Immunity-Triggering Effector from the Barley Smut Fungus Ustilago hordei Resides in an Ustilaginaceae-Specific Cluster Bearing Signs of Transposable Element-Assisted Evolution

    KAUST Repository

    Ali, Shawkat

    2014-07-03

    The basidiomycete smut fungus Ustilago hordei was previously shown to comprise isolates that are avirulent on various barley host cultivars. Through genetic crosses we had revealed that a dominant avirulence locus UhAvr1 which triggers immunity in barley cultivar Hannchen harboring resistance gene Ruh1, resided within an 80-kb region. DNA sequence analysis of this genetically delimited region uncovered the presence of 7 candidate secreted effector proteins. Sequence comparison of their coding sequences among virulent and avirulent parental and field isolates could not distinguish UhAvr1 candidates. Systematic deletion and complementation analyses revealed that UhAvr1 is UHOR_10022 which codes for a small effector protein of 171 amino acids with a predicted 19 amino acid signal peptide. Virulence in the parental isolate is caused by the insertion of a fragment of 5.5 kb with similarity to a common U. hordei transposable element (TE), interrupting the promoter of UhAvr1 and thereby changing expression and hence recognition of UhAVR1p. This rearrangement is likely caused by activities of TEs and variation is seen among isolates. Using GFP-chimeric constructs we show that UhAvr1 is induced only in mated dikaryotic hyphae upon sensing and infecting barley coleoptile cells. When infecting Hannchen, UhAVR1p causes local callose deposition and the production of reactive oxygen species and necrosis indicative of the immune response. UhAvr1 does not contribute significantly to overall virulence. UhAvr1 is located in a cluster of ten effectors with several paralogs and over 50% of TEs. This cluster is syntenous with clusters in closely-related U. maydis and Sporisorium reilianum. In these corn-infecting species, these clusters harbor however more and further diversified homologous effector families but very few TEs. This increased variability may have resulted from past selection pressure by resistance genes since U. maydis is not known to trigger immunity in its corn host.

  10. Proteomic Analysis of the Relationship between Metabolism and Nonhost Resistance in Soybean Exposed to Bipolaris maydis.

    Science.gov (United States)

    Dong, Yumei; Su, Yuan; Yu, Ping; Yang, Min; Zhu, Shusheng; Mei, Xinyue; He, Xiahong; Pan, Manhua; Zhu, Youyong; Li, Chengyun

    2015-01-01

    Nonhost resistance (NHR) pertains to the most common form of plant resistance against pathogenic microorganisms of other species. Bipolaris maydis is a non-adapted pathogen affecting soybeans, particularly of maize/soybean intercropping systems. However, no experimental evidence has described the immune response of soybeans against B. maydis. To elucidate the molecular mechanism underlying NHR in soybeans, proteomics analysis based on two-dimensional polyacrylamide gel electrophoresis (2-DE) was performed to identify proteins involved in the soybean response to B. maydis. The spread of B. maydis spores across soybean leaves induced NHR throughout the plant, which mobilized almost all organelles and various metabolic processes in response to B. maydis. Some enzymes, including ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), mitochondrial processing peptidase (MPP), oxygen evolving enhancer (OEE), and nucleoside diphosphate kinase (NDKs), were found to be related to NHR in soybeans. These enzymes have been identified in previous studies, and STRING analysis showed that most of the protein functions related to major metabolic processes were induced as a response to B. maydis, which suggested an array of complex interactions between soybeans and B. maydis. These findings suggest a systematic NHR against non-adapted pathogens in soybeans. This response was characterized by an overlap between metabolic processes and response to stimulus. Several metabolic processes provide the soybean with innate immunity to the non-adapted pathogen, B. maydis. This research investigation on NHR in soybeans may foster a better understanding of plant innate immunity, as well as the interactions between plant and non-adapted pathogens in intercropping systems.

  11. 玉米瘤黑粉病抗性鉴定技术的评价%Evaluation of Technique for Resistance Identification on Ustilago maydis

    Institute of Scientific and Technical Information of China (English)

    石菁; 张金文; 陆继有

    2010-01-01

    选用西北灌区生产上具有代表性的10个玉米品种,采用不同接种方法及接种浓度在玉米不同生育时期人工接种玉蜀黍黑粉菌,研究抗性鉴定技术,筛选出一整套重复性好、发病完全、受外界因素干扰小的接种技术体系.结果表明,供试玉米品种接种玉蜀黍黑粉菌的最佳方法是注射法,最佳接种时期为6叶1心期,最佳接种浓度为9×10~3~10×10~3个孢子/mL.

  12. Chemical Control Techniques of Ustilago Maydis in Maize%玉米瘤黑粉病的化学防治技术

    Institute of Scientific and Technical Information of China (English)

    车宏伟; 车庆成; 董海

    2010-01-01

    通过拌种与田间药剂喷雾防治玉米瘤黑粉病试验,认为拌种处理较田间药剂喷雾防效明显提高.三唑类杀菌剂拌种防效在61.87%~73.11%,喷雾处理防效在40.65%~52.17%,明显高于苯并咪唑类杀菌剂多菌灵的防效.

  13. STUDY ON FUNGICIDE CONTROL OF USTILAGO MAYDIS IN MAIZE%玉米瘤黑粉病药剂防治研究

    Institute of Scientific and Technical Information of China (English)

    李晓丽; 李凤岭; 臧少先; 李贺年; 张义奇

    2002-01-01

    通过室内筛选和田间试验,结果表明,稀唑醇为防治玉米瘤黑粉病较好的药剂,在玉米心叶末期撒稀唑醇与辛硫磷复配的颗粒剂,对玉米瘤黑粉病和玉米螟均有较好的防治效果,防效分别为75.7%和85.0%.

  14. Influence of Stenocarpella maydis infected corn on the composition of corn kernel and its conversion into ethanol

    Science.gov (United States)

    Widespread epidemics of Stenocarpella ear rot (formerly Diplodia ear rot) have occurred throughout the central U.S. Corn Belt in recent years, but the influence of S. maydis infected grain on corn ethanol production is unknown. In this study, S. maydis infected ears of variety 'Heritage 4646' were h...

  15. Przenoszenie się grzyba Ustilago perennans Bostr. z nasionami rajgrasu wyniosłego [Transmission of Ustilago perennans Rostr. with tall oat grass seeds

    Directory of Open Access Journals (Sweden)

    J. W. Tomala-Bednarek

    2015-06-01

    Full Text Available It was established that the mycelium of Ustilago perennans does not grow into the pericarp and seed coat of Arrhenatherum elatius caryopses. The main source of seedling infection at this year .seeds sowing were mycelium and gemmes present in hull tissues whereas ait last-year seeds sowing-the spores wintering on the hull and caryopsis surfaces, as they proved to be more survived. The ability of the fungus to seedling infection was decreasing gradually with the seed ageing.

  16. Ecological genetics of the Bromus tectorum (Poaceae) - Ustilago Bullata (Ustilaginaceae): A role for frequency dependent selection?

    Science.gov (United States)

    Susan E. Meyer; David L. Nelson; Suzette Clement; Alisa Ramakrishnan

    2010-01-01

    Evolutionary processes that maintain genetic diversity in plants are likely to include selection imposed by pathogens. Negative frequency-dependent selection is a mechanism for maintenance of resistance polymorphism in plant - pathogen interactions. We explored whether such selection operates in the Bromus tectorum - Ustilago bullata pathosystem. Gene-for-gene...

  17. Dicty_cDB: Contig-U02109-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2 Dormant Teliospore Ustilago maydis cDN... 46 1.1 1 ( CF645338 ) K35_C07 Filamentous Forced Diploid Ustilag...o maydi... 46 1.1 1 ( CF644499 ) K21_H02 Filamentous Forced Diploid Ustilago mayd

  18. Allozyme-specific modification of a maize seed chitinase by a protein secreted by the fungal pathogen Stenocarpella maydis.

    Science.gov (United States)

    Naumann, Todd A; Wicklow, Donald T

    2010-07-01

    Stenocarpella maydis causes both dry-ear rot and stalk rot of maize. Maize inbred lines have varying levels of resistance to ear rot caused by S. maydis. The genetic basis of resistance appears to rely on multiple genetic factors, none of which are known. The commonly used stiff-stalk inbred line B73 has been shown to be strongly susceptible to ear rot caused by S. maydis. Here, we report that the ChitA protein alloform from B73, ChitA-F, encoded by a known allele of the chiA gene, is susceptible to modification by a protein (Stm-cmp) secreted by S. maydis. We also identify a new allele of chiA (from inbred line LH82) which encodes ChitA-S, an alloform of ChitA that is resistant to Stm-cmp modification. Chitinase zymogram analysis of seed from a commercial field showed the presence of both ChitA alloforms in healthy ears, and showed that ChitA-F but not ChitA-S was modified in ears rotted by S. maydis. The ChitA-F protein was purified from inbred line B73 and ChitA-S from LH82. ChitA-F was modified more efficiently than ChitA-S by S. maydis protein extracts in vitro. The chiA gene from LH82 was cloned and sequenced. It is a novel allele that encodes six polymorphisms relative to the known allele from B73. This is the first demonstration that the susceptibility to modification of a fungal targeted plant chitinase differs among inbred lines. These findings suggest that the LH82 chiA allele may be a specific genetic determinant that contributes to resistance to ear rot caused by S. maydis whereas the B73 allele may contribute to susceptibility.

  19. Dicty_cDB: FC-IC0794 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 75 |CF643875.1 K13_F09 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA s...equence. 40 9e-04 2 CF639788 |CF639788.1 D19_G01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA se... 40 0.001 2 CF642755 |CF642755.1 D55_F01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. ...10 |CF639410.1 D15_D02 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 40 0.001 2 dna upd

  20. Dicty_cDB: SFI878 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available |CF644841.1 K26_B01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA seq... 4.4 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 4....5 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced Diploid Ustilago maydis cDNA ...3', mRNA sequence. 34 4.5 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced Diploid Ustilago maydis cDNA 3',

  1. Dicty_cDB: CHE109 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ce. 32 2.2 2 CF644841 |CF644841.1 K26_B01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence.... 34 5.7 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced Diploid Ustilago maydis... cDNA 3', mRNA sequence. 34 7.6 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced Diploid Ustilago maydis cD...NA 3', mRNA sequence. 34 7.7 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced Diploid Ustilago maydis cDNA

  2. Damages caused by Bipolaris maydis in Panicum maximum cv. Tanzânia Dano causado por Bipolaris maydis em Panicum maximum cv. Tanzânia

    Directory of Open Access Journals (Sweden)

    Gilmar Franzener

    2010-12-01

    Full Text Available The aim of this research was to evaluate the damage caused by Bipolaris maydis in the quantity and quality of the Tanzania grass production. Tanzania grass plants were cultivated in pots of PVC tubes (50 cm of height x 15 cm of diameter containing a mixture of soil and sand (3:1. The plants were inoculated with spore pathogen suspension containing 102, 104 and 106 conidia/mL to obtain different levels of disease. Severity and number of tillers were evaluated weekly. After four weeks, it were evaluated the weight of the fresh matter of leaves (FM, percentage of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF and acid detergent fiber (ADF. The disease reduced significantly the plant tillering and FM after the second evaluation (P O objetivo deste trabalho foi avaliar o dano causado por Bipolaris maydis (helmintosporiose na quantidade e qualidade da produção do capim Tanzânia. Plantas de capim Tanzânia foram cultivadas em vasos de tubos de PVC (50 cm de altura x 15 cm de diâmetro contendo mistura solo/areia (3:1. As plantas foram inoculadas com suspensões de esporos do patógeno com diferentes concentrações (102, 104 e 106 conídios/mL, visando obter gradiente de severidade de manchas foliares. Foram realizadas avaliações semanais da severidade e do número de perfilhos. Após quatro semanas avaliou-se o peso da matéria fresca de folhas (MF, porcentagem de matéria seca (PMS, de proteína bruta (PB, de fibra em detergente neutro (FDN e de fibra em detergente ácido (FDA. A doença reduziu significativamente o perfilhamento e MF a partir da segunda avaliação (P < 0,05. Não houve correlação significativa entre as variáveis PMS, FDA e área abaixo da curva de progresso da doença (AACPD. No entanto, houve correlação positiva (P < 0,01 e negativa (P < 0 ,05 entre PB e FDN, respectivamente, com a AACPD, como possível resultado da atividade do patógeno. Estes resultados indicam que B. maydis inibe o desenvolvimento do

  3. Induced Resistance by the Toxin Filtrate of Bipolaris maydis Race T Cultivation

    Institute of Scientific and Technical Information of China (English)

    MA Chun-hong; ZHAI Cai-xia; WANG Li-an; CHEN Xia; LI Yun-chao; GUO Xiu-lin; CUI Si-ping; LI Guang-min

    2006-01-01

    Resistance to maize southern leaf spot disease was induced by the low-concentration filtrate of Bipolaris maydis race T cultivation in an experiment. The nuclear neterogeny corn C103 was used as the test material. The lesion area on the leaves was significant difference by connalysis. The lesion areas on pretreated leaves were (0.3 ± 0.05)- (0.9 ± 0.5) mm2, but those on the control were (23.1 ± 8.7) mm2. At the same time, the changes in peroxidase, phenylalanine, ammonialyase, and malondialdehvde activities were determined. During 0-96 h of inspection, phenylalanine and ammonialyase (PAL) activities increased by 64.2%, peroxidase (POD) activities increased by 41.2%, but the malondialdenvde (MDA) content decreased by 29.7% compared with the control. It seems that the low-concentration filtrate of Bipolaris maydis race T cultivation itself can be used as an elicitor to enhance the induced resistance.

  4. Resistance in barley against Drechslera teres induced by Bipolaris maydis and Septoria nodorum

    DEFF Research Database (Denmark)

    Jørgensen, Hans Jørgen Lyngs; Lobeck, P.S.; Thordal-Christensen, Hans

    1998-01-01

    pre-treated with conidial suspensions of either of the two non-barley pathogens Bipolaris maydis from maize or Septoria nodorum from wheat. The results suggested that induced resistance was involved, but detailed studies of the mechanisms responsible were not carried out. Such investigations are...... for inhibiting D. teres in barley after inoculation with B. maydis and S. nodorum. This was done by quantitative histological examination of the primary infection stages of D. teres, by qualitative studies of the later development of this fungus, and by studies of the expression of defence response genes...... in the host after inoculation with B. maydis and S. nodorum. Materials and methods B. maydis and S. nodorum were applied to leaves of the barley cultivar Lenka 24 h before D. teres. The primary infection processes of D. teres were investigated by light microscopy of epidermal strips made from the first leaf 3...

  5. Observations on the attacks of Ustilago avenae (Pers. Jens. fungus on oat

    Directory of Open Access Journals (Sweden)

    Viorel POPESCU

    1967-08-01

    Full Text Available In vorliegender Arbeit wird von den Verfasser eine vergleichende Untersuchung uber den Infektionsgrad von Flugbard (Ustilago avenae Pers. Jens. bei 10 Hafersorten and-arten nach den Methoden Zade, Reed und einer originellen halbanaturlichen Methode geboten. Aus den erhaltenen Daten geht hervor, dass die wirksamste Methode der kunstlichen Infektion die Zade'sche ist, gefolgt von der Methodenach Reed. Man nimmt an, dass die besseren Ergebnisse durch die Methode nach Zade auf die Verwengund von Regenwasser bei der Herstellung der Sporensuspension zuruckgefuhrt warden konnen; welckes sich als ein gunstigeres Medium fur die Keimung erweist. Als die widerstandsfahigsten Sorten gegen diese Pflanzenkrankheit haben, sich Richland und Markton Oast 166 erwiesen, wahrend die Art Avena nuda und Avena sativa mit den Sorten Peragis und Baragan 878 am anfalligsten waren.

  6. A Novel Glycosylphosphatidylinositol-Anchored Glycoside Hydrolase from Ustilago esculenta Functions in β-1,3-Glucan Degradation

    OpenAIRE

    Nakajima, Masahiro; Yamashita, Tetsuro; Takahashi, Machiko; Nakano, Yuki; Takeda, Takumi

    2012-01-01

    A glycoside hydrolase responsible for laminarin degradation was partially purified to homogeneity from a Ustilago esculenta culture filtrate by weak-cation-exchange, strong-cation-exchange, and size-exclusion chromatography. Three proteins in enzymatically active fractions were digested with chymotrypsin followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, resulting in the identification of three peptide sequences that shared significant similarity to a putative β-1...

  7. Aggressiveness between genetic groups I and II of isolates of Cercospora zeae-maydis Agressividade entre isolados dos grupos genéticos I e II de Cercospora zeae-maydis

    Directory of Open Access Journals (Sweden)

    Sandra Marisa Mathioni

    2006-12-01

    Full Text Available For many years, the gray leaf spot disease (GLS caused by the fungus Cercospora zeae-maydis Tehon & Daniels, was not considered an important pathogen of maize (Zea mays, L. in Brazil. However, the recent adoption of agronomical practices such as no-tillage and cultivation under central pivot irrigation systems increased the incidence and severity to the extent that GLS is now one of the most important diseases of maize. Isolates of C. zeae-maydis can be distinguished by two genetic groups (I and II based on AFLP markers and on polymorphisms of the ITS and 5.8S rDNA regions. Until now, however, the biological implications of this distinction remain unclear. This study investigated whether isolates from the two genetic groups differ in aggressiveness towards maize. For this, symptoms of a susceptible hybrid were evaluated under greenhouse conditions with 9 and 11 isolates of C. zeae-maydis from groups I and II, respectively. Plants in the V3 growth stage were inoculated by placing sorghum seeds colonized with the pathogen in the leaf whorl and symptoms were evaluated with a visual rating scale 30 days later. On average, isolates of genetic group II were more aggressive than those of group I, with mean disease scores of 3.1 and 2.3, respectively. Differences were also observed between experiments, which suggested that group I and II might also differ in their fitness under different environments. This is the first report on differences in aggressiveness between the two genetic groups of C. zeae-maydis.Durante muitos anos, a cercosporiose, causada pelo fungo Cercospora zeae-maydis Tehon & Daniels, não foi considerada importante para a cultura do milho (Zea mays, L. no Brasil. Entretanto, a recente utilização de práticas culturais como o plantio direto e o cultivo sob pivôs centrais favoreceram o aumento de sua severidade e incidência, de forma que a doença é hoje considerada uma das mais importantes da cultura. Isolados de C. zeae-maydis

  8. Dicty_cDB: VSF854 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available |CF643875.1 K13_F09 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 40 0.001 2 CF639788 ...|CF639788.1 D19_G01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequ... |CF639410.1 D15_D02 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 40 0.001 2 CF644777 ...|CF644777.1 K25_C11 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 40 0.001 2 dna update...5.1 D55_F01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 40

  9. Dicty_cDB: VHA155 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available e 1061005864011 3', genomic survey sequence. 46 1.6 1 DQ767612 |DQ767612.1 Triticum aestivum clone BAC TA105...go maydis genomic clone 1204168, genomic survey sequence. 44 6.2 1 DX756422 |DX756422.1 2365937 VV03 Ustilag...o maydis genomic clone 1025158, genomic survey sequence. 44 6.2 1 DX733440 |DX733...440.1 2315736 VV03 Ustilago maydis genomic clone 1006812, genomic survey sequence. 44 6.2 1 DX722040 |DX7220...40.1 2310775 VV03 Ustilago maydis genomic clone 1002065, genomic survey sequence. 44 6.2 1 DX660090 |DX66009

  10. Agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of Cercospora zeae-maydis.

    Science.gov (United States)

    Lu, Yuanyuan; Xiao, Shuqin; Wang, Fen; Sun, Jiaying; Zhao, Likun; Yan, Libin; Xue, Chunsheng

    2017-02-01

    An efficient Agrobacterium tumefaciens-mediated transformation (ATMT) approach was developed for the plant pathogenic fungus, Cercospora zeae-maydis, which is the causative agent of gray leaf spot in maize. The transformation was evaluated with five parameters to test the efficiencies of transformation. Results showed that spore germination time, co-cultivation temperature and time were the significant influencing factors in all parameters. Randomly selected transformants were confirmed and the transformants were found to be mitotically stable, with single-copy T-DNA integration in the genome. T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. Thus, the ATMT approach is an efficient tool for insertional mutagenesis of C. zeae-maydis.

  11. 原子力显微镜研究萎锈灵对玉米黑粉菌细胞壁的影响%AFM Study Effects of carboxin on the wall of Ustilago maydis

    Institute of Scientific and Technical Information of China (English)

    古宁宇; 刘彩胜; 唐纪琳

    2011-01-01

    用原子力显微镜研究了不同浓度萎锈灵药物对玉米黑粉菌表面的形貌的影响。结果发现黑粉菌经过萎锈灵作用后表面出现很多无规则的小坑,通过表征统计表明菌体坑的数目随着药物浓度增加而增加,而菌体坑的深度没有改变。对萎锈灵作用玉米黑粉菌的影响有了进一步的认识。

  12. Analysis of Gene Structure, Cloning and Expression of cyp51 from Ustilago maydis%玉米黑粉菌cyp51基因结构分析与克隆表达

    Institute of Scientific and Technical Information of China (English)

    韩睿; 邓灵福; 黎晨; 张青叶; 张劼; 高强; 熊丽; 万坚; 刘德立

    2008-01-01

    通过生物信息学手段分析cyp51基因结构,并根据GenBank登记的玉米黑粉茼cyp51 DNA序列,设计cyp51引物和两对分别截短不同跨膜区的突变体引物,构建了多种重组表达质粒及突变体重组表达质粒.选用不同宿主菌包括Escherichia coli BL21(DE3).BL21(DE3)pLysS和Rosetta(DE3)诱导表达并优化条件.SDS-PAGE分析结果表明:只有突变体pET32-YH-35能够在E. coli BL21(DE3)中高效表达(30℃,0.5 mmol/L IPRG诱导).通过与戊唑醇等4种商品化杀菌剂农药和14种XF系列农药先导化合物的紫外结合光谱分析表明:重组蛋白具有生物学活性.其中一种XF系列化合物的结合常数接近商品化杀菌剂,有可能开发为新的杀菌剂,为设计开发新型高效抗真菌新药提供了理论依据.

  13. 玉米黑粉菌cyp51基因上游调控区克隆及生物信息学分析%Cloning and sequence analysis of upstream region of cyp51 gene from Ustilago maydis

    Institute of Scientific and Technical Information of China (English)

    黎晨; 张华; 韩睿; 袁永泽; 杨江科; 刘德立

    2009-01-01

    为探明玉米黑粉菌cyp51基因的表达调控机制,根据玉米黑粉菌cyp51基因cDNA的5'-序列,采用染色体步移技术,获得其5'-上游调控区序列,总长为490bp.利用NNPP分析软件预测转录起始位点,并采用TFSEARCH 1.3软件分析转录因子结合位点.结果显示:转录起始位点位于上游134bp处;上游调控区不仅包含启动子的核心结构序列TATA盒(分别位于-30、-58、-318和-348 bp处)和CAAT盒(分别位于-150、-161和-191 bp处),亦包含多个转录因子结合位点,如AP-4、GATA-1、CdxA、Dfd和Oct-1等;在上游调控序列中嘌呤含量高,而且从-222 bp处开始存在4个连续高嘌呤含量的热激转录因子特异性结合位点(HSE).

  14. Computational prediction on subcellular location of proteins encoded in Ustilago maydis genome%玉米黑粉菌基因组编码蛋白作用位点的计算机预测

    Institute of Scientific and Technical Information of China (English)

    黄婉; 范成明; 吴毅歆; 何月秋

    2008-01-01

    利用巴公布的玉米黑粉病菌基因组全序列数据及信号肽预测软件SignalP v3.0、亚细胞器中蛋白定位分布预测软件Tar-getP v1.01、跨膜螺旋结构预测软件TMHMM v2.0和膜锚定位点预测软件Big-PI Predictor预测分析了玉米黑粉病菌基因组编码蛋白的作用位点.结果表明.在6522个ORF中,具有分泌功能的有543个,占全基因组基因总数的8.3%;作用位点在线粒体的有1552个,占全基因组基因总数的23.4%;具有跨膜结构的有1269个,占全基因组基因总数的19.5%;锚定在膜上的有56个,占全基因组基园总数的0.9%.

  15. Regulation of stomatal tropism and infection by light in Cercospora zeae-maydis: evidence for coordinated host/pathogen responses to photoperiod?

    Directory of Open Access Journals (Sweden)

    Hun Kim

    2011-07-01

    Full Text Available Cercospora zeae-maydis causes gray leaf spot of maize, which has become one of the most widespread and destructive diseases of maize in the world. C. zeae-maydis infects leaves through stomata, which is predicated on the ability of the pathogen to perceive stomata and reorient growth accordingly. In this study, the discovery that light was required for C. zeae-maydis to perceive stomata and infect leaves led to the identification of CRP1, a gene encoding a putative blue-light photoreceptor homologous to White Collar-1 (WC-1 of Neurospora crassa. Disrupting CRP1 via homologous recombination revealed roles in multiple aspects of pathogenesis, including tropism of hyphae to stomata, the formation of appressoria, conidiation, and the biosynthesis of cercosporin. CRP1 was also required for photoreactivation after lethal doses of UV exposure. Intriguingly, putative orthologs of CRP1 are central regulators of circadian clocks in other filamentous fungi, raising the possibility that C. zeae-maydis uses light as a key environmental input to coordinate pathogenesis with maize photoperiodic responses. This study identified a novel molecular mechanism underlying stomatal tropism in a foliar fungal pathogen, provides specific insight into how light regulates pathogenesis in C. zeae-maydis, and establishes a genetic framework for the molecular dissection of infection via stomata and the integration of host and pathogen responses to photoperiod.

  16. Pheromone-Related Inhibitors of Ustilago hordei Mating and Tilletia tritici Teliospore Germination.

    Science.gov (United States)

    Kosted, Paula J; Gerhardt, Shirley A; Sherwood, John E

    2002-02-01

    ABSTRACT Ustilago hordei, the causal agent of barley covered smut, produces mating pheromones that break down to smaller peptide compounds that act as potent inhibitors of mating and germination in several fungi. The pheromones are members of the farnesylated family of proteins. Synthetic peptide analogs of the pheromone derivatives, ranging in size from 4 mers to full length pheromones, were farnesylated, methyl esterified, or both and tested for mating or teliospore germination inhibition with U. hordei or Tilletia tritici, respectively. N-Acetyl-S-farnesylcysteine, which inhibits processing of Ras, and other sulfur-containing compounds such as homocysteine or methionine, were likewise modified and tested. The most potent inhibitors were methionine methyl ester and modified 4-mer peptides from both pheromones. Alanine scanning of the inhibitory 4 mers determined that the native amino acid sequence was specific for a high level of activity. The sulfur amino acids appear to be required for inhibition. Glasshouse studies using selected antagonists of mating and teliospore germination as seed treatments inhibited covered smut of barley and common bunt of wheat, although the level of control was inconsistent. The use of pheromone-related antagonists to mating or teliospore germination is a promising, novel strategy for control of smut and bunt diseases.

  17. Glycoproteins from sugarcane plants regulate cell polarity of Ustilago scitaminea teliospores.

    Science.gov (United States)

    Millanes, Ana-María; Fontaniella, Blanca; Legaz, María-Estrella; Vicente, Carlos

    2005-03-01

    Saccharum officinarum, cv. Mayarí, is a variety of sugarcane resistant to smut disease caused by Ustilago scitaminea. Sugarcane naturally produces glycoproteins that accumulate in the parenchymatous cells of stalks. These glycoproteins contain a heterofructan as polysaccharide moiety. The concentration of these glycoproteins clearly increases after inoculation of sugarcane plants with smut teliospores, although major symptoms of disease are not observed. These glycoproteins induce homotypic adhesion and inhibit teliospore germination. When glycoproteins from healthy, non-inoculated plants are fractionated, they inhibit actin capping, which occurs before teliospore germination. However, inoculation of smut teliospores induce glycoprotein fractions that promote teliospore polarity and are different from those obtained from healthy plants. These fractions exhibit arginase activity, which is strongly enhanced in inoculated plants. Arginase from healthy plants binds to cell wall teliospores and it is completely desorpted by sucrose, but only 50% of arginase activity from inoculated plants is desorpted by the disaccharide. The data presented herein are consistent with a model of excess arginase entry into teliospores. Arginase synthesized by sugarcane plants as a response to the experimental infection would increase the synthesis of putrescine, which impedes polarization at concentration values higher than 0.05 mM. However, smut teliospores seem to be able to change the pattern of glycoprotein production by sugarcane, thereby promoting the synthesis of different glycoproteins that activate polarization after binding to their cell wall ligand.

  18. Resistance in barley against Drechslera teres induced by Bipolaris maydis and Septoria nodorum

    DEFF Research Database (Denmark)

    Jørgensen, Hans Jørgen Lyngs; Lobeck, P.S.; Thordal-Christensen, Hans

    1998-01-01

    not been examined to any great extent, with a few exceptions such as the rice-Pyricularia oryzae system. In a previous report [1], the severity of barley net blotch caused by the necrotrophic pathogen Drechslera teres and hyphal growth in the host tissue was shown to be strongly reduced when leaves were...... pre-treated with conidial suspensions of either of the two non-barley pathogens Bipolaris maydis from maize or Septoria nodorum from wheat. The results suggested that induced resistance was involved, but detailed studies of the mechanisms responsible were not carried out. Such investigations are...... in the host after inoculation with B. maydis and S. nodorum. Materials and methods B. maydis and S. nodorum were applied to leaves of the barley cultivar Lenka 24 h before D. teres. The primary infection processes of D. teres were investigated by light microscopy of epidermal strips made from the first leaf 3...

  19. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

    Science.gov (United States)

    Petatán-Sagahón, Iván; Anducho-Reyes, Miguel Angel; Silva-Rojas, Hilda Victoria; Arana-Cuenca, Ainhoa; Tellez-Jurado, Alejandro; Cárdenas-Álvarez, Isabel Oyuki; Mercado-Flores, Yuridia

    2011-01-01

    Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease. PMID:22016606

  20. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

    Directory of Open Access Journals (Sweden)

    Yuridia Mercado-Flores

    2011-08-01

    Full Text Available Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.

  1. Isolation of bacteria with antifungal activity against the phytopathogenic fungi Stenocarpella maydis and Stenocarpella macrospora.

    Science.gov (United States)

    Petatán-Sagahón, Iván; Anducho-Reyes, Miguel Angel; Silva-Rojas, Hilda Victoria; Arana-Cuenca, Ainhoa; Tellez-Jurado, Alejandro; Cárdenas-Álvarez, Isabel Oyuki; Mercado-Flores, Yuridia

    2011-01-01

    Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.

  2. Development of a Stigma Maydis Sanitarian Tea Bag%玉米须保健袋泡茶的研制

    Institute of Scientific and Technical Information of China (English)

    瞿颖; 蒋立勤; 李喜; 张舟艺; 姜宇翔; 丁力

    2011-01-01

    The aim of this work is to develop a kind of sanitarian tea bag using Stigma Maydis as well as green tea, liquorice, hawthorn and lotus leaf. The antioxidative ability of the tea bag was also analyzed. The Stigma Maydis Sanitarian tea bag was prepared by decotion, extraction,spraying and malaxating. The optimum technology parameters were obtained through orthogonal analysis as well as sensory perception analysis.Total content of flavone was determined by aluminum colorimetry assay. For antioxidant activity, DPPH radical scavenging activity and reducing power of the product were evaluated. The optimum compositions was as following: 69% Stigma Maydis, 20% green tea, 7.0% hawthorn, 3.7%lotus leaf and 0.7% liquorice. High level of total flavone content and strong antioxidation ability was found in the product. The Stigma Maydis tea bag contains functional contents including flavone and suggests good health effects.%开发以玉米须和绿茶为主料,以甘草、山楂、荷叶为辅料的玉米须保健袋泡荼,并评价其抗氧化能力.采用煎煮、提取、喷揉法制备玉米须袋泡茶,采用正交法、感官评价法得到袋泡茶最佳工艺参数,采用铝比色法测定总黄酮含量,采用DPPH自由基清除法和总还原力法测定产品的抗氧化能力.最佳配料配方中每份含玉米须69%,绿茶20%、山楂7.0%、荷叶3.7%、甘草0.7%,产品总黄酮含量较高并且具有良好的抗氧化能力.玉米须袋泡荼中含有黄酮等功效成分具有较好保健效果.

  3. cDNA-AFLP analysis reveals that maize resistance to Bipolaris maydis is associated with the induction of multiple defense-related genes

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The fungal pathogen Bipolaris maydis invades by direct penetration into maize leaf veins. In order to understand the resistance mechanism of maize to B. maydis strain 523, cDNA-AFLP (amplified fragment length polymorphism) analysis was conducted to compare the changes in mRNA transcripts in response to B. maydis infection between a highly disease-resistant (HDR) line and a susceptible (S) line. 13 cDNA fragments derived from the genes showing enhanced expression after fungal infection, named HDR genes, were isolated from the HDR line. Northern blot analysis showed that 5 HDR genes were induced by fungal infection in the HDR, but not the S lines. The 5 HDR genes showed homology to previously characterized genes involved in disease resistance. A full-length HDR10 cDNA was isolated. It had a capacity to encode a protein of 284 amino acids. The deduced amino acid sequence of the HDR10 gene was homologous to a fungal infection-induced protein from Cicer arietinum and a hypersensitive response protein from maize, respectively. These results suggest that maize resistance to B. maydis infection in the HDR line may be mediated by the induction of multiple defense-related genes.``

  4. Genetic diversity ofUstilago hordei in Tibetan areas as revealed by RAPD and SSR

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yu; CHAO Gui-mei; LIU Jia-jia; ZHU Ming-qi; WANG Yang; FENG Bai-li

    2016-01-01

    Covered smut, which is caused byUstilago hordei(Pers.) Lagerh., is one of the most damaging diseases of highland barley (Hordeum vulgare Linn. var. nudum Hook. f) in Tibetan areas of China. To understand the molecular diversity ofU. hordei, a total of 27 isolates, which were colected from highland barley plants from Tibet, Sichuan, Qinghai, and Gansu provinces/autonomous region, were analyzed using random ampliifed polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Among the 100 RAPD primers used, 24 primers exhibited polymorphism. A total of 111 fragments were ampliifed, of which 103 were polymorphic with a polymorphic rate of 92.79%. The average observed number of aleles (Na), effective number of aleles (Ne), Nei’s genetic diversity (H), Shannon’s information index (I) and polymorphism information content (PIC) value in the RAPD markers were 1.9279, 1.5016, 0.2974, 0.4503 and 0.6428, respectively. For the SSR markers, 40 of the 111 primer pairs exhibited polymorphism and provided a total of 119 bands, of which 109 were polymorphic and accounted for 91.60% of the total bands. TheNa,Ne,H,I andPIC values of the SSR markers were 1.9160, 1.4639, 0.2757, 0.4211 and 0.4340, respectively. The similarity coefifcients ranged from 0.4957 to 0.9261 with an average of 0.7028 among al the 27 isolates used. The dendrogram, which was developed based on the RAPD and SSR combined marker dataset showed that the 27U. hordei isolates were divided into 3 clusters at similarity coefifcient of 0.7314. We determined that RAPD and SSR markers can be successfuly used to assess the genetic variation amongU. hordei isolates. The RAPD markers revealed higher levels of genetic polymorphism than did the SSR markers in this study. There existed a moderate genetic difference among isolates. The molecular variation and differentiation was somewhat associated with geographical origin but not for al of the isolates.

  5. Analyses of expressed sequence tags from the maize foliar pathogen Cercospora zeae-maydis identify novel genes expressed during vegetative, infectious, and reproductive growth

    Directory of Open Access Journals (Sweden)

    Kema Gert HJ

    2008-11-01

    Full Text Available Abstract Background The ascomycete fungus Cercospora zeae-maydis is an aggressive foliar pathogen of maize that causes substantial losses annually throughout the Western Hemisphere. Despite its impact on maize production, little is known about the regulation of pathogenesis in C. zeae-maydis at the molecular level. The objectives of this study were to generate a collection of expressed sequence tags (ESTs from C. zeae-maydis and evaluate their expression during vegetative, infectious, and reproductive growth. Results A total of 27,551 ESTs was obtained from five cDNA libraries constructed from vegetative and sporulating cultures of C. zeae-maydis. The ESTs, grouped into 4088 clusters and 531 singlets, represented 4619 putative unique genes. Of these, 36% encoded proteins similar (E value ≤ 10-05 to characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions and biological processes based on Gene Ontology (GO classification. We identified numerous, previously undescribed genes with potential roles in photoreception, pathogenesis, and the regulation of development as well as Zephyr, a novel, actively transcribed transposable element. Differential expression of selected genes was demonstrated by real-time PCR, supporting their proposed roles in vegetative, infectious, and reproductive growth. Conclusion Novel genes that are potentially involved in regulating growth, development, and pathogenesis were identified in C. zeae-maydis, providing specific targets for characterization by molecular genetics and functional genomics. The EST data establish a foundation for future studies in evolutionary and comparative genomics among species of Cercospora and other groups of plant pathogenic fungi.

  6. Effecf of pH and some cations on activity of acid phosphatase secreted from Ustilago sp. isolated from acid sulphate soil

    Directory of Open Access Journals (Sweden)

    Chairatana Nilnond

    2007-03-01

    Full Text Available Acid phosphatase secreted from Ustilago sp. is able to hydrolyze organic phosphorus. These soil yeast microorganisms were isolated from rice roots grown in acid sulphate soil that generally contains highamount of aluminum (Al, iron (Fe and manganese (Mn ions. Therefore, the objectives of this study were to examine the effect of pH and some cations on acid phosphatase activity. Two isolates of Ustilago sp., AR101and AR102, were cultured in 100 mL of modified Pikovskaya's broth containing Na-phytate, pH 4, and acid phosphatase activity was determined at pH 2.0-7.0. Effect of Al, Fe, and Mn, including calcium (Ca ions,on growth of AR101 and AR102, secreted acid phosphatase activity, and the ability of acid phosphatase on the phosphorus release from Na-phytate by Ustilago sp. were investigated. It was found that the optimum pH for acid phosphatase activity was 3.5-4.5. The activity of acid phosphatase secreted from AR101 (3,690nmol min-1 mL-1 was remarkably higher than that from AR102 (956 nmol min-1 mL-1. Aluminum, iron, manganese and calcium ions in the medium did not affect the growth of either isolate. The activity of secretedacid phosphatase of AR101 was inhibited by Al and Ca ion, and synthesis of acid phosphatase of Ustilago sp. AR102 was possibly stimulated by Fe ion. Both AR101 and AR102 solubilized Na-phytate, resulting in therelease of P. However, some amount of released P was then precipitated with Al and Fe ions as the highly insoluble Fe- or Al- phosphate.

  7. Dicty_cDB: CFJ713 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available uence. 42 2.3 1 CF644841 |CF644841.1 K26_B01 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequen...ce. 34 2.8 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced Diploid Ustilago may...dis cDNA 3', mRNA sequence. 34 3.8 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced Diploid Ustilago maydis... cDNA 3', mRNA sequence. 34 3.8 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced Diploid Ustilago maydis cD

  8. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica

    National Research Council Canada - National Science Library

    Bailey, A M; Mena, G L; Herrera-Estrella, L

    1991-01-01

    ...) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000...

  9. Isomerization mechanism of aspartate to isoaspartate implied by structures of Ustilago sphaerogena ribonuclease U2 complexed with adenosine 3'-monophosphate.

    Science.gov (United States)

    Noguchi, Shuji

    2010-07-01

    Aspartates in proteins are isomerized non-enzymatically to isoaspartate via succinimide in vitro and in vivo. In order to elucidate the mechanism of isoaspartate formation within the Asp45-Glu46 sequence of Ustilago sphaerogena ribonuclease U2 based on three-dimensional structure, crystal structures of ribonuclease U2 complexed with adenosine 3'-monophosphate have been solved at 0.96 and 0.99 A resolution. The crystal structures revealed that the C(gamma) atom of Asp45 is located just beside the main-chain N atom of Glu46 and that the conformation which is suitable for succinimide formation is stabilized by a hydrogen-bond network mediated by water molecules 190, 219 and 220. These water molecules are suggested to promote the formation of isoaspartate via succinimide: in the succinimide-formation reaction water 219 receives a proton from the N atom of Glu46 as a general base and waters 190 and 220 stabilize the tetrahedral intermediate, and in the succinimide-hydrolysis reaction water 219 provides a proton for the N atom of Glu46 as a general acid. The purine-base recognition scheme of ribonuclease U2 is also discussed.

  10. Effects of temperature, light regime and substrates on the production and germination of Stenocarpella maydis pycnidiospores=Efeito de temperatura, regime de luminosidade e substratos sobre a produção e germinação de picnidioporos de Stenocarpella maydis

    Directory of Open Access Journals (Sweden)

    Jonathan Marcel Bolzan

    2012-01-01

    Full Text Available This study aimed to evaluate the production and germination of Stenocarpella maydis pycnidiospores under in vitro conditions as affected by substrate composition (sorghum, wheat, black oat or barley, light regime (continuous dark, 12-h light dark-1 or continuous light conditions, and incubation temperature (21, 24, 27, 30 or 33ºC. Each substrate (20 g of grain was soaked in 100 mL of water for 24h and sterilized twice for 20 min at 127ºC. Three plugs (5 mm diameter of a single-spored culture of S. maydis were used as inocula for each substrate. Assessments of pycnidiospore production per gram of grain and percent germination were made 14 days after inoculation. Barley, black oat or wheat grains were the best substrates for the mass production of S. maydis pycnidiospores and the maintenance of high germination rates. The highest pycnidiospore production (67,600 pycnidiospores g-1 was obtained using barley grain as a substrate with incubation at 27ºC under a 12-h light dark-1 cycle.. A produção e a germinação de picnidiosporos de Stenocarpella maydis foram avaliadas in vitro sob a influência da composição de substratos (sorgo, trigo, aveia preta e cevada, regime de luminosidade (escuro contínuo, fotoperíodo de 12h ou luz contínua e temperatura de inbucação (21, 24, 27, 30 ou 33ºC. Três discos de 5 mm de cultura monospórica de S. maydis foram inoculados em cada substrato. A produção e o percentual de germinação de picnidiosporos foram avaliados aos 14 dias após a inoculação. Grãos de cevada, aveia preta e trigo foram os melhores substratos na produção e germinação de picnidiosporos de S. maydis. A maior concentração (67.600 picnidiosporos g-1 foi obtida em grãos de cevada, à 27ºC e 12h de fotoperíodo.

  11. The KP4 killer protein gene family

    Science.gov (United States)

    Killer protein 4 (KP4) is a well studied toxin secreted by the maize smut fungus Ustilago maydis that kills sensitive Ustilago strains as well as inhibits Fusarium and plant root growth. This small, cysteine rich protein is encoded by a virus that depends on host survival for replication. KP4 functi...

  12. Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut

    Science.gov (United States)

    The corn smut fungus, Ustilago maydis, is a global pathogen responsible for extensive agricultural losses. Control of corn smut using traditional breeding has met with limited success because natural resistance to U. maydis is organ specific and involves numerous maize genes. Here, we present a tran...

  13. Dicty_cDB: Contig-U12127-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ultiple Tissues, Normal... 58 2e-05 2 ( CF638362 ) D01_G08 Filamentous Forced Dip...tilago maydis genomic clone 813735... 48 2e-05 2 ( CF638424 ) D02_E01 Filamentous Forced..... 48 2e-05 2 ( CF639153 ) D11_H09 Filamentous Forced Diploid Ustilago maydi... 48 2e-05 2 ( AC128812 ) Ratt

  14. La marchitez tardía del maíz (Zea mays L.) causada por Cephalosporium maydis en la Península Ibérica, y otros hongos asociados

    OpenAIRE

    Carmen Maria Ortiz-Bustos; García-Carneros, Ana B.; Leire Molinero-Ruiz

    2015-01-01

    Las especies de hongos de suelo asociadas a Cephalosporium maydis como agente causal de la marchitez tardía del maíz en la Península Ibérica se identificaron muestreando 19 campos con síntomas de marchitez en las principales zonas de cultivo entre 2011 y 2012. En el 47% de los campos no se identificó C. maydis, pero sí Fusarium graminearum, F. verticillioides, F. equiseti, F. proliferatum, Macrophomina phaseolina, Rhizoctonia solani y Trichoderma harzianum infectando las plantas de maíz. En l...

  15. Xylanases, Cellulases, and Acid Protease Produced by Stenocarpella maydis Grown in Solid-state and Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Edna María Hernández-Domínguez

    2014-03-01

    Full Text Available Activity levels of extracellular hydrolytic enzymes produced by Stenocarpella maydis, a fungal pathogen of maize, have so far not been reported. Production of xylanase, cellulase, and acid protease by this ascomycete using different culture media in solid-state and submerged fermentation was studied. In solid-state fermentation, polyurethane foam was used as an inert support, and corncob, corn leaves, and broken corn were used as biodegradable supports. The highest xylanase activity was produced in the medium with xylan in both fermentation systems, reaching 18,020 U/L and 19,266 U/L for submerged and solid-state fermentation, respectively. Cellulase production was observed only in the culture medium with carboxymethylcellulose, obtaining values of 7,872 U/L in submerged fermentation and 9,439 U/L in solid-state fermentation. The acid protease was produced only in minimal medium with glucose in acidic pH, reaching the highest levels of activity in SSF (806 U/L. The corncob was the best biodegradable support for the production of xylanases and acid protease. Two isoenzymes of xylanase and cellulase were observed in both fermentation systems, and three isoenzymes of xylanase were produced on the biodegradable supports.

  16. Occurrence of Bipolaris maydis leaf spot on tanzania guineagrass in the north region of the Mato Grosso state

    Directory of Open Access Journals (Sweden)

    Tauan Rimoldi Tavanti

    2016-08-01

    Full Text Available The monoculture associated with the intensification of livestock results on appearance of diseases in forages, which can lead to significant losses. Symptomatic leaves of Tanzania guineagrass (Panicum maximum collected in Alta Floresta and Nova Guarita, Mato Grosso State, Brazil, were analyzed in the Plant Pathology Laboratory of UNEMAT/Alta Floresta. Fragments of diseased tissues previously disinfected in 70% ethanol and sodium hypochlorite to 1000 ppm solutions, were plated in a potato dextrose agar culture medium. The plates were stored at 25 °C and 12-hours photo period, for seven days. After this period Tanzânia guineagrass (Panicum maximum plants were inoculated with the pathogen from infected plants collected on both cities. To complete Koch's postulate, after the onset of symptoms, the pathogen was reisolated. The fungus Bipolaris maydis, causal agent of leaf spot was identified, based on the observation of fungal structures in the light microscope, the use of sort keys and the of Koch’s postulate.

  17. Safety Evaluation, in Vitro and in Vivo Antioxidant Activity of the Flavonoid-Rich Extract from Maydis stigma

    Directory of Open Access Journals (Sweden)

    Ke-Zheng Peng

    2015-12-01

    Full Text Available This study aimed to assess the acute toxicity and safety of flavonoid-rich extract from Maydis stigma (FMS in mice. The in vitro antioxidant activity of FMS was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH and 2,2′-azinobis-(3-ethyl-benzthiazoline-6-sulphonate (ABTS scavenging assays. Furthermore, the in vivo antioxidant of FMS against ethanol-induced oxidative damage in mice was determined by analysis of the serum total superoxide dismutase (T-SOD activity, malondialdehyde (MDA content, liver tissue glutathione (GSH content, and protein carbonyl (PC content in liver tissue. The oral administration of FMS at doses of 30 g/kg did not cause death in mice, and there were no significant biologically adverse effects in mice. These results indicated that the median lethal dose (LD50 is higher than this dose. The IC50 values of FMS for the DPPH and ABTS scavenging activity were 50.73 and 0.23 mg/mL, respectively. Meanwhile, FMS could significantly enhance T-SOD activity, reduce MDA content in the serum, increase GSH content, and decrease PC content in the liver tissue at the tested doses (25, 50, 100, 200 mg/kg·day. These results indicate that FMS can be generally regarded as safe and used potentially as a bioactive source of natural antioxidants.

  18. A novel glycosylphosphatidylinositol-anchored glycoside hydrolase from Ustilago esculenta functions in β-1,3-glucan degradation.

    Science.gov (United States)

    Nakajima, Masahiro; Yamashita, Tetsuro; Takahashi, Machiko; Nakano, Yuki; Takeda, Takumi

    2012-08-01

    A glycoside hydrolase responsible for laminarin degradation was partially purified to homogeneity from a Ustilago esculenta culture filtrate by weak-cation-exchange, strong-cation-exchange, and size-exclusion chromatography. Three proteins in enzymatically active fractions were digested with chymotrypsin followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, resulting in the identification of three peptide sequences that shared significant similarity to a putative β-1,3-glucanase, a member of glucoside hydrolase family 16 (GH16) from Sporisorium reilianum SRZ2. A gene encoding a laminarin-degrading enzyme from U. esculenta, lam16A, was isolated by PCR using degenerate primers designed based on the S. reilianum SRZ2 β-1,3-glucanase gene. Lam16A possesses a GH16 catalytic domain with an N-terminal signal peptide and a C-terminal glycosylphosphatidylinositol (GPI) anchor peptide. Recombinant Lam16A fused to an N-terminal FLAG peptide (Lam16A-FLAG) overexpressed in Aspergillus oryzae exhibited hydrolytic activity toward β-1,3-glucan specifically and was localized both in the extracellular and in the membrane fractions but not in the cell wall fraction. Lam16A without a GPI anchor signal peptide was secreted extracellularly and was not detected in the membrane fraction. Membrane-anchored Lam16A-FLAG was released completely by treatment with phosphatidylinositol-specific phospholipase C. These results suggest that Lam16A is anchored in the plasma membrane in order to modify β-1,3-glucan associated with the inner cell wall and that Lam16A is also used for the catabolism of β-1,3-glucan after its release in the extracellular medium.

  19. Obtención de jarabe a partir del almidón del maíz morado Zea Maydis L.

    OpenAIRE

    Lancho Ruíz, Ana Celina

    2015-01-01

    El maíz morado Zea maydis L., es un cereal nativo del Perú, que es apreciado por su contenido de antocianinas en la coronta. Sin embargo los granos tienen un contenido de almidón del 16.09% (Lancho, 2009). En los laboratorios de la Facultad de Ingeniería Pesquera y de Alimentos de la Universidad Nacional del Callao , y en la Facultad de Ciencias Biológicas de la Universidad Nacional Mayor de San Marcos, se extrajo el almidón y evaluó la hidrolisis química, también la hidrolisis enzimática; co...

  20. Preliminary Trial of 11 New Hybrid Maize Genotype to The Resistance on Java Downy Mildew (Peronosclerospora maydis

    Directory of Open Access Journals (Sweden)

    Budi Setyawan

    2016-04-01

    Full Text Available Maize or corn (Zea mays L. belongs to the family of grasses (Poaceae.  Maize is grown globally and one of the most important cereal crop in the world.  In many countries, corn is the main agricultural crop, and are used as food, feed and industrial raw materials. Together with rice and wheat, corn included in the cereals that provide about 65% carbohydrates and 50% protein that humans need. For this purpose, many developing countries, especially in Asia and Africa are in a strong effort to increase their corn yields through the use of better seeds.  Although in Indonesia, corn is the second important food crop after rice, however, with the rapid growth of the livestock industry, corn is a major component (60% in feed ingredient.  It is estimated that more than 55% of the corn used for feed in Indonesia.  Java Downy Mildew (Peronosclerospora maydis is the main disease that is concerned by maize corn growers. This disease often resulting in substantial losses for farmers, even reach 100% in susceptible genotypes.  Therefore trial on 11 new prospective hybrids was conducted with the expectation that they can be classified in the criteria “less resistant or higher” according to the criteria of modified Reid (2005.  Of the 11 new hybrids, 3 new hybrids (27.3% classified in the criteria “very resistant”, 4 new hybrids (36.4% classified in the criteria “resistant”, 2 new hybrid (18.2% classified in the criteria “less resistant”, and 2 other new hybrids (18.2% classified in the criteria “less susceptible”.

  1. La marchitez tardía del maíz (Zea mays L. causada por Cephalosporium maydis en la Península Ibérica, y otros hongos asociados

    Directory of Open Access Journals (Sweden)

    Carmen Maria Ortiz-Bustos

    2015-06-01

    Full Text Available Las especies de hongos de suelo asociadas a Cephalosporium maydis como agente causal de la marchitez tardía del maíz en la Península Ibérica se identificaron muestreando 19 campos con síntomas de marchitez en las principales zonas de cultivo entre 2011 y 2012. En el 47% de los campos no se identificó C. maydis, pero sí Fusarium graminearum, F. verticillioides, F. equiseti, F. proliferatum, Macrophomina phaseolina, Rhizoctonia solani y Trichoderma harzianum infectando las plantas de maíz. En los campos restantes, junto a C. maydis se identificaron otros hongos de suelo en porcentajes apreciables: F. verticillioides (19%, F. proliferatum (19%, F.equiseti (9%, F. oxysporum (9% y Pythium oligandrum (9%. El crecimiento vascular de C. maydis y de otras especies fúngicas en plantas de maíz se confirmó analizando plantas con marchitez procedentes de tres campos diferentes. Tanto C. maydis como F. graminearum, F. equiseti, F. proliferatum y T. harzianum se aislaron de la inserción entre la raíz y tallo y a 10 cm de altura en el tallo de las plantas. El efecto de la infección por C. maydis sobre la producción de las plantas de maíz se cuantificó en macetas y condiciones seminaturales en el 2011. En plantas inoculadas se obtuvo una reducción del peso de las mazorcas del 54%, además de pesos de raíz y de parte aérea (tallo y hojas significativamente menores en comparación con el control no inoculado, lo que sugiere el gran impacto económico que puede tener la marchitez tardía en condiciones naturales. Asimismo este trabajo pone de manifiesto el grado de complejidad de la etiología de la marchitez tardía, que debería ser estudiado mediante la confirmación de la patogenicidad de los hongos de suelo identificados en maíz, con el fin de determinar el papel que puede jugar cada una de estas especies en el desarrollo de la enfermedad y/o severidad de los síntomas.

  2. Study on the Biological Characteristics of Spore of Ustilago maydis and Virulence of 3 kinds of Fungicide Against it in Lab.%玉米瘤黑粉菌冬孢子生物学特性及三种杀菌剂对其萌发的影响

    Institute of Scientific and Technical Information of China (English)

    李广领; 陈锡岭; 秦雪峰; 郭彦亮

    2007-01-01

    通过对玉米瘤黑粉菌冬孢子的生物学特性的研究,明确了该冬孢子萌发的最适温度为25~30 ℃,在自然光照射的条件下萌发较好.用清水浸泡3 d,再用0.025 mol/L盐酸处理24 h能显著促进玉米瘤黑粉病菌冬孢子萌发.以天门冬氨酸和硫酸铵为培养液,它们分别在质量分数为2×10-5时玉米瘤黑粉菌冬孢子萌发率最高,分别达81.4%和78.1%.用三唑酮,烯唑醇,包衣杀在田间推荐剂量下对冬孢子进行处理,结果表明,它们对冬孢子的萌发率有明显的抑制作用.

  3. Dicty_cDB: SFF749 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ans cDNA clone yk11a11 : 5' end, single read. 42 3.7 1 CF644841 |CF644841.1 K26_B01 Filamentous Forced Diplo...id Ustilago maydis cDNA 3', mRNA sequence. 34 6.8 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced Diploid ...Ustilago maydis cDNA 3', mRNA sequence. 34 9.1 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced... Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 9.1 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced

  4. Dicty_cDB: SFD890 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available BOGZM14TR BOGZ Brassica oleracea genomic clone BOGZM14, DNA sequence. 42 2.7 1 CF644841 |CF644841.1 K26_B01 Filamentous Forced...1582.1 D41_E10 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 5.0 2 CF643189 |CF64318...9.1 D60_G10 Filamentous Forced Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 5.0 2 CF643884 |CF643884.1 K13_G06 Filamentous Forc...ed Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 5.2 2 dna update 2003.12.23 H

  5. Membrane Stabilization and Detoxification of Acetaminophen-Mediated Oxidative Onslaughts in the Kidneys of Wistar Rats by Standardized Fraction of Zea mays L. (Poaceae, Stigma maydis

    Directory of Open Access Journals (Sweden)

    S. Sabiu

    2016-01-01

    Full Text Available This study evaluated membrane stabilization and detoxification potential of ethyl acetate fraction of Zea mays L., Stigma maydis in acetaminophen-induced oxidative onslaughts in the kidneys of Wistar rats. Nephrotoxic rats were orally pre- and posttreated with the fraction and vitamin C for 14 days. Kidney function, antioxidative and histological analyses were thereafter evaluated. The acetaminophen-mediated significant elevations in the serum concentrations of creatinine, urea, uric acid, sodium, potassium, and tissue levels of oxidized glutathione, protein-oxidized products, lipid peroxidized products, and fragmented DNA were dose-dependently assuaged in the fraction-treated animals. The fraction also markedly improved creatinine clearance rate, glutathione, and calcium concentrations as well as activities of superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase in the nephrotoxic rats. These improvements may be attributed to the antioxidative and membrane stabilization activities of the fraction. The observed effects compared favorably with that of vitamin C and are informative of the fraction’s ability to prevent progression of renal pathological conditions and preserve kidney functions as evidently supported by the histological analysis. Although the effects were prominently exhibited in the fraction-pretreated groups, the overall data from the present findings suggest that the fraction could prevent or extenuate acetaminophen-mediated oxidative renal damage via fortification of antioxidant defense mechanisms.

  6. Membrane Stabilization and Detoxification of Acetaminophen-Mediated Oxidative Onslaughts in the Kidneys of Wistar Rats by Standardized Fraction of Zea mays L. (Poaceae), Stigma maydis

    Science.gov (United States)

    Sabiu, S.; O'Neill, F. H.

    2016-01-01

    This study evaluated membrane stabilization and detoxification potential of ethyl acetate fraction of Zea mays L., Stigma maydis in acetaminophen-induced oxidative onslaughts in the kidneys of Wistar rats. Nephrotoxic rats were orally pre- and posttreated with the fraction and vitamin C for 14 days. Kidney function, antioxidative and histological analyses were thereafter evaluated. The acetaminophen-mediated significant elevations in the serum concentrations of creatinine, urea, uric acid, sodium, potassium, and tissue levels of oxidized glutathione, protein-oxidized products, lipid peroxidized products, and fragmented DNA were dose-dependently assuaged in the fraction-treated animals. The fraction also markedly improved creatinine clearance rate, glutathione, and calcium concentrations as well as activities of superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase in the nephrotoxic rats. These improvements may be attributed to the antioxidative and membrane stabilization activities of the fraction. The observed effects compared favorably with that of vitamin C and are informative of the fraction's ability to prevent progression of renal pathological conditions and preserve kidney functions as evidently supported by the histological analysis. Although the effects were prominently exhibited in the fraction-pretreated groups, the overall data from the present findings suggest that the fraction could prevent or extenuate acetaminophen-mediated oxidative renal damage via fortification of antioxidant defense mechanisms. PMID:27579048

  7. Dicty_cDB: SFC144 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available protein, complete cds. 331 2e-87 1 CF644841 |CF644841.1 K26_B01 Filamentous Forced...in WCFS1 complete genome; segment 9/11. 30 4.4 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced Diploid Ust...ilago maydis cDNA 3', mRNA sequence. 34 4.6 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced Diploid Ustila...go maydis cDNA 3', mRNA sequence. 34 4.6 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced

  8. Dicty_cDB: CHA733 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ans cosmid F21H12, complete sequence. 42 3.2 1 CF644841 |CF644841.1 K26_B01 Filamentous Forced...antarum strain WCFS1 complete genome; segment 9/11. 30 5.3 2 CF641582 |CF641582.1 D41_E10 Filamentous Forced... Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 6.9 2 CF643189 |CF643189.1 D60_G10 Filamentous Forced... Diploid Ustilago maydis cDNA 3', mRNA sequence. 34 6.9 2 CF643884 |CF643884.1 K13_G06 Filamentous Forced

  9. Mapping and validation of quantitative trait loci for resistance to Cercospora zeae-maydis infection in tropical maize (Zea mays L.).

    Science.gov (United States)

    Pozar, Gilberto; Butruille, David; Silva, Heyder Diniz; McCuddin, Zoe Patterson; Penna, Julio Cesar Viglioni

    2009-02-01

    Breeding for resistance to gray leaf spot, caused by Cercospora zeae-maydis (Cz) is paramount for many maize environments, in particular under warm and humid growing conditions. In this study, we mapped and characterized quantitative trait loci (QTL) involved in the resistance of maize against Cz. We confirmed the impact of the QTL on disease severity using near-isogenic lines (NILs), and estimated their effects on three major agronomic traits using their respective near isogenic hybrids (NIHs), which we obtained by crossing the NILs with an inbred from a complementary heterotic pool. We further validated three of the four QTL that were mapped using the Multiple Interval Mapping approach and showed LOD values>2.5. NILs genotype included all combinations between favorable alleles of the two QTL located in chromosome 1 (Q1 in bin 1.05 and Q2 in bin 1.07), and the allele in chromosome 3 (Q3 in bin 3.07). Each of the three QTL separately significantly reduced the severity of Cz. However, we found an unfavorable epistatic interaction between Q1 and Q2: presence of the favorable allele at one of the QTL allele effectively nullified the effect of the favorable allele at the other. In contrast, the interaction between Q2 and Q3 was additive, promoting the reduction of the severity to a greater extent than the sum of their individual effects. When evaluating the NIH we found significant individual effects for Q1 and Q3 on gray leaf spot severity, for Q2 on stalk lodging and grain yield, and for Q3 on grain moisture and stalk lodging. We detected significant epitasis between Q1 and Q2 for grain moisture and between Q1 and Q3 for stalk lodging. These results suggest that the combination of QTL impacts the effectiveness of marker-assisted selection procedures in commercial product development programs.

  10. Kinetics of α-amylase and α-glucosidase inhibitory potential of Zea mays Linnaeus (Poaceae), Stigma maydis aqueous extract: An in vitro assessment.

    Science.gov (United States)

    Sabiu, S; O'Neill, F H; Ashafa, A O T

    2016-05-13

    Corn silk (Zea mays L., Stigma maydis) is an important herb used traditionally in many parts of the world to treat array of diseases including diabetes mellitus. Inhibitors of α-amylase and α-glucosidase offer an effective strategy to modulate levels of post prandial hyperglycaemia via control of starch metabolism. This study evaluated α-amylase and α-glucosidase inhibitory potentials of corn silk aqueous extract. Active principles and antioxidant attributes of the extract were also analysed. The α-amylase inhibitory potential of the extract was investigated by reacting its different concentrations with α-amylase and starch solution, while α-glucosidase inhibition was determined by pre-incubating α-glucosidase with different concentrations of the extract followed by addition of p-nitrophenylglucopyranoside. The mode(s) of inhibition of the enzymes were determined using Lineweaver-Burke plot. In vitro analysis of the extract showed that it exhibited potent and moderate inhibitory potential against α-amylase and α-glucosidase, respectively. The inhibition was concentration-dependent with respective half-maximal inhibitory concentration (IC50) values of 5.89 and 0.93mg/mL. Phytochemical analyses revealed the presence of alkaloids, flavonoids, phenols, saponins, tannins and phytosterols as probable inhibitory constituents. Furthermore, the extract remarkably scavenges reactive oxygen species like DPPH and nitric oxide radicals, elicited good reducing power and a significant metal chelating attributes. Overall, the non-competitive and uncompetitive mechanism of action of corn silk extract is due to its inhibitory effects on α-amylase and α-glucosidase, respectively. Consequently, this will reduce the rate of starch hydrolysis, enhance palliated glucose levels, and thus, lending credence to hypoglycaemic candidature of corn silk. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  11. Dicty_cDB: Contig-U11652-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1 ( DX636193 ) 2106994 VV02 Ustilago maydis genomic clone 812537... 48 0.79 1 ( EH028505 ) MMN_43_E01 Haploid cells grown in Minimal... Media m... 48 0.79 1 ( EH028097 ) MMN_38_H05 Haploid cells grown in Minimal Media m

  12. The cAMP Signaling and MAP Kinase Pathways in Plant Pathogenic Fungi

    NARCIS (Netherlands)

    Mehrabi, R.; Zhao, X.; Kim, Y.; Xu, J.R.

    2009-01-01

    The key components of the well conserved cyclic AMP signaling and MAP kinase pathways have been functionally characterized in the corn smut Ustilago maydis, rice blast fungus Magnaporthe grisea, and a few other fungal pathogens. In general, the cAMP signaling and the MAP kinase cascade homologous to

  13. Ubc2, an Ortholog of the Yeast Ste50p Adaptor, Possesses a Basidiomycete-Specific Carboxy terminal Extension Essential for Pathogenicity Independent of Pheromone Response.

    Science.gov (United States)

    Proteins involved in the MAP kinase pathway controlling mating, morphogenesis and pathogenicity have been identified previously in the fungus Ustilago maydis. One of these, the Ubc2 adaptor protein, possesses a basidiomycete-specific structure. In addition to containing SAM and RA domains typical of...

  14. Influence of brown stink bug feeding, planting date and sampling time on common smut infection of maize

    Science.gov (United States)

    Phytopathogen infections are frequently influenced by both biotic and abiotic factors in a crop field. The effect of brown stink bug, Euschistus servus (Hemiptera: Pentatomidae), feeding and planting date and sampling time on common smut (Ustilago maydis) infection percentage of maize plants was exa...

  15. Dss1 interaction with Brh2 as a regulatory mechanism for recombinational repair

    DEFF Research Database (Denmark)

    Zhou, Qingwen; Kojic, Milorad; Cao, Zhimin;

    2007-01-01

    Brh2, the BRCA2 ortholog in Ustilago maydis, enables recombinational repair of DNA by controlling Rad51 and is in turn regulated by Dss1. Interplay with Rad51 is conducted via the BRC element located in the N-terminal region of the protein and through an unrelated domain, CRE, at the C terminus...

  16. Kinesin-3 and dynein cooperate in long-range retrograde endosome motility along a nonuniform microtubule array

    NARCIS (Netherlands)

    Schuster, M.; Kilaru, S.; Fink, G.; Collemare, J.A.R.; Roger, Y.; Steinberg, G.

    2011-01-01

    The polarity of microtubules (MTs) determines the motors for intracellular motility, with kinesins moving to plus ends and dynein to minus ends. In elongated cells of Ustilago maydis, dynein is thought to move early endosomes (EEs) toward the septum (retrograde), whereas kinesin-3 transports them to

  17. On the antifungal mode of action of tridemorph

    NARCIS (Netherlands)

    Kerkenaar, A.; Barug, D.; Kaars Sijpesteijn, A.

    1979-01-01

    Tridemorph (2,6-dimethyl-N-tridecylmorpholine) was active against representative of nearly all taxonomic groups of fungi; gram-positive bacteria were also sensitive although gram-negative were not. Tridemorph, 3–10 μg/ml, inhibited the multiplication of sporidia of Ustilago maydis more strongly than

  18. Kinesin-3 and dynein cooperate in long-range retrograde endosome motility along a nonuniform microtubule array

    NARCIS (Netherlands)

    Schuster, M.; Kilaru, S.; Fink, G.; Collemare, J.A.R.; Roger, Y.; Steinberg, G.

    2011-01-01

    The polarity of microtubules (MTs) determines the motors for intracellular motility, with kinesins moving to plus ends and dynein to minus ends. In elongated cells of Ustilago maydis, dynein is thought to move early endosomes (EEs) toward the septum (retrograde), whereas kinesin-3 transports them to

  19. On the antifungal mode of action of tridemorph

    NARCIS (Netherlands)

    Kerkenaar, A.; Barug, D.; Kaars Sijpesteijn, A.

    1979-01-01

    Tridemorph (2,6-dimethyl-N-tridecylmorpholine) was active against representative of nearly all taxonomic groups of fungi; gram-positive bacteria were also sensitive although gram-negative were not. Tridemorph, 3–10 μg/ml, inhibited the multiplication of sporidia of Ustilago maydis more strongly than

  20. Identification of the antagonistic bacterium YB01against Bipolaris maydis and its antagonism%玉蜀黍平脐蠕孢拮抗细菌YB01的鉴定及拮抗作用

    Institute of Scientific and Technical Information of China (English)

    韩梅; 彭帅; 依艳丽

    2011-01-01

    通过形态特征、生理生化及16S rDNA同源性序列分析对玉蜀黍平脐蠕孢(Bipolaris maydis)拮抗细菌YB01进行鉴定,结果表明,该菌为苍白杆菌属的一种(Ochrobactrum sp.),序列号为HQ141339.YB01对玉蜀黍平脐蠕孢的拮抗作用研究表明:YB01培养液可以有效抑制玉蜀黍平脐蠕孢的生长、孢子的形成与萌发.显微镜观察结果表明,用YB01培养液处理的玉蜀黍平脐蠕孢菌丝体膨大变形,细胞质发生浓缩,表明YB01的拮抗作用主要是通过其代谢产物影响玉蜀黍平脐蠕孢细胞壁而实现的.%The antagonistic bacterium YB01 was identified by the characteristics of morphology, physiology, biochemical tests and sequence analysis of 16S rDNA, which was identified as Ochrobactrum sp. The antifungal mechanism of the YB01 was investigated in this study. The results indicated that mycelial growth, spore formation and germination of Bipolaris maydis could be inhibited by the metabolites. The mycelia affected by the metabolites also showed surface nodulation and protoplasmic condensation. It is speculated that the metabolites may target at the fungal cell walls.

  1. 三种杀菌剂对玉米褐斑病菌的毒力及田间控制作用%Toxicity and field control efficacy of three fungicides against to Physoderma maydis

    Institute of Scientific and Technical Information of China (English)

    李俊虎; 姜兴印; 戈大庆; 王燕; 段强; 王冲; 鲍静

    2011-01-01

    采用孢子萌发法,分别测定了代森锰锌、戊唑醇和噁霉灵对玉米褐斑病菌pysoderma maydis的室内毒力;并通过田间小区试验,研究了80%代森锰锌可湿性粉剂(WP)、430 g/L戊唑醇悬浮剂(SC)和70%噁霉灵可湿性粉剂(WP)对病原菌致病力的影响,以及在8叶期茎叶喷雾处理对该病害的田间控制作用.结果表明:代森锰锌对病原菌休眠孢子囊的萌发具有明显的抑制作用,EC50值为66.71 mg/L,而戊唑醇和噁霉灵的抑制效果较差;代森锰锌和戊唑醇能有效降低病原菌的致病力,处理小区植株发病叶片均控制在穗位叶以下,且穗位叶叶绿素含量、净光合速率和根系活力均明显高于对照区,防效分别达91.31%和88.99%;代森锰锌和戊唑醇处理小区玉米增产效果明显,增产率分别达13.8%和11.5%.%The spore germination method was used to determine the toxicities of tebuconazole,mancozeb and hymexazol to Physoderma maydis, and field trials were conducted to ascertain the effects of mancozeb 800 WP,430 g/L tebuconazole SC and hymexazol 700 WP on pathogenicity, as well as control efficacy when applied at 8-leaf stage of corn by spray. The results showed that mancozeb exhibited great inhibition to the germination of the resting sporangia of P. maydis, with the EC5o value of 66.71 mg/L, while tebuconazole and hymexazol had little effects. The pathogenicity was effectively lowered by the treatments of mancozeb and tebuconazole, which suppressed the occurrence of corn brown spot below the ear leaf and increased the content of chlorophyll, net photosynthetic and root activity. The control efficacy of mancozeb and tebuconazole against P. maydis were 91.31% and 88.99% respectively,which resulted in an increased yields of corn by 13.8% and 11.5%,comparing with that of CK.

  2. Antifungal activity of a virally encoded gene in transgenic wheat.

    Science.gov (United States)

    Clausen, M; Kräuter, R; Schachermayr, G; Potrykus, I; Sautter, C

    2000-04-01

    The cDNA encoding the antifungal protein KP4 from Ustilago maydis-infecting virus was inserted behind the ubiquitin promoter of maize and genetically transferred to wheat varieties particularly susceptible to stinking smut (Tilletia tritici) disease. The transgene was integrated and inherited over several generations. Of seven transgenic lines, three showed antifungal activity against U. maydis. The antifungal activity correlated with the presence of the KP4 transgene. KP4-transgenic, soil-grown wheat plants exhibit increased endogenous resistance against stinking smut.

  3. Mating Type Loci of Sporisorium reilianum: Novel Pattern with Three a and Multiple b Specificities

    OpenAIRE

    Schirawski, Jan; Heinze, Bernadette; Wagenknecht, Martin; Kahmann, Regine

    2005-01-01

    Sporisorium reilianum and Ustilago maydis are two closely related smut fungi, which both infect maize but differ fundamentally in their mode of plant invasion and site of symptom development. As a prelude to studying the molecular basis of these differences, we have characterized the mating type loci of S. reilianum. S. reilianum has two unlinked mating type loci, a and b. Genes in both loci and adjacent regions show a high degree of synteny to the corresponding genes of U. maydis. The b locu...

  4. Study on Biological Characteristics of Cercospora zeae-maydis in Southwest China%西南地区玉米灰斑病菌生物学特性研究

    Institute of Scientific and Technical Information of China (English)

    崔丽娜; 张小飞; 邹成佳; 李晓; 杨晓蓉; 罗怀海

    2012-01-01

    为了进一步防治玉米灰斑病奠定基础,对西南地区玉蜀黍尾孢菌的生物学特性进行了初步研究.结果表明,该病菌生长温度为10 ~35℃,最适生长温度为20~25℃;病原菌对酸碱度的适应范围较广,在pH 4 -11都能生长,以pH 5 ~7为最适pH值;分生孢子萌发温度为5 ~35℃,最适温度为20 ~25℃;光照对菌丝和孢子萌发无显著影响;病原菌最适碳源为葡萄糖,最适氮源为酵母青,致死温度为55℃(10 min).%Some biological characters of Cercospora zeae-maydis were studied in this experiment. The results showed that the pathogen could grow and develop between 10 and 35 ℃ , and the optimum temperature was between 20 and 25 ℃, For growth of the pathogen, there had no strict pH requirement: the favorable pH was 4 -11 and the optimum pH was 5-7. For germination of conidia, the favorable temperature for conidial germination was between 5 and 35 ℃and the optimum temperature was 20 -25 ℃. Effect of light was not obvious on mycelial growth and conidial germination. The optimum carbon source was glucose, and the optimum nitrogen source was yeast extract. The lethal temperature for pathogen was at 55 ℃ for 10 minutes.

  5. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica.

    OpenAIRE

    Bailey, A. M.; Mena, G L; Herrera-Estrella, L

    1991-01-01

    Phytophthora capsici and P.parasitica were transformed to hygromycin B resistance using plasmids pCM54 and pHL1, which contain the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000. Transformation frequencies of over 500 transformants per micrograms of DNA per ...

  6. Pathogenicity determinants in smut fungi revealed by genome comparison.

    Science.gov (United States)

    Schirawski, Jan; Mannhaupt, Gertrud; Münch, Karin; Brefort, Thomas; Schipper, Kerstin; Doehlemann, Gunther; Di Stasio, Maurizio; Rössel, Nicole; Mendoza-Mendoza, Artemio; Pester, Doris; Müller, Olaf; Winterberg, Britta; Meyer, Elmar; Ghareeb, Hassan; Wollenberg, Theresa; Münsterkötter, Martin; Wong, Philip; Walter, Mathias; Stukenbrock, Eva; Güldener, Ulrich; Kahmann, Regine

    2010-12-10

    Biotrophic pathogens, such as the related maize pathogenic fungi Ustilago maydis and Sporisorium reilianum, establish an intimate relationship with their hosts by secreting protein effectors. Because secreted effectors interacting with plant proteins should rapidly evolve, we identified variable genomic regions by sequencing the genome of S. reilianum and comparing it with the U. maydis genome. We detected 43 regions of low sequence conservation in otherwise well-conserved syntenic genomes. These regions primarily encode secreted effectors and include previously identified virulence clusters. By deletion analysis in U. maydis, we demonstrate a role in virulence for four previously unknown diversity regions. This highlights the power of comparative genomics of closely related species for identification of virulence determinants.

  7. Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut.

    Science.gov (United States)

    Allen, Aron; Islamovic, Emir; Kaur, Jagdeep; Gold, Scott; Shah, Dilip; Smith, Thomas J

    2011-10-01

    The corn smut fungus, Ustilago maydis, is a global pathogen responsible for extensive agricultural losses. Control of corn smut using traditional breeding has met with limited success because natural resistance to U. maydis is organ specific and involves numerous maize genes. Here, we present a transgenic approach by constitutively expressing the Totivirus antifungal protein KP4, in maize. Transgenic maize plants expressed high levels of KP4 with no apparent negative impact on plant development and displayed robust resistance to U. maydis challenges to both the stem and ear tissues in the greenhouse. More broadly, these results demonstrate that a high level of organ independent fungal resistance can be afforded by transgenic expression of this family of antifungal proteins.

  8. The maize cystatin CC9 interacts with apoplastic cysteine proteases.

    Science.gov (United States)

    van der Linde, Karina; Mueller, André N; Hemetsberger, Christoph; Kashani, Farnusch; van der Hoorn, Renier A L; Doehlemann, Gunther

    2012-11-01

    In a recent study we identified corn cystain9 (CC9) as a novel compatibility factor for the interaction of the biotrophic smut fungus Ustilago maydis with its host plant maize. CC9 is transcriptionally induced during the compatible interaction with U. maydis and localizes in the maize apoplast where it inhibits apoplastic papain-like cysteine proteases. The proteases are activated during incompatible interaction and salicylic acid (SA) treatment and, in turn, are sufficient to induce SA signaling including PR-gene expression. Therefore the inhibition of apoplastic papain-like cysteine proteases by CC9 is essential to suppress host immunity during U. maydis infection. Here were present new experimental data on the cysteine protease-cystatin interaction and provide an in silco analysis of plant cystatins and the identified apoplastic cysteine proteases.

  9. Optimized Conditions of Botway Solid Fermentation for Transforming Inonotus Obliquus with Stigma Maydis%桦褐孔菌与玉米须双向固体发酵工艺优化研究

    Institute of Scientific and Technical Information of China (English)

    曹思思; 郑宏宇; 陈丽艳; 方自若; 王伟明

    2014-01-01

    Objective:To research the cultivating condition on bothway solid fermentation conditions of Inonotus obliquus for transforming Chinese medicine Stigma maydis in order to increase the conversion rate .Method:The fermentation products were achieved by solid fermentation method .The average growth rate ,the content of total polysaccharides and total triterpenoid were observed respectively by mycelium growth rate and by ultravio -let spectroscopy , at last use the weighted average method to determine the best cultivating condition .Results:The best cultivating condition , the cultivating temperature ( 28 ±2 )℃, the best ratio of solid material of Stig-ma and Corn cob was 1∶3, substrates of dry weight and wet weight were 90g/bag.The liquid inoculation a-mount was 2ml/bag and culturing circle was 50d.Conclusion:Using bothway solid fermentation condition tech-nology ,through different proportion solid medium to obtain HXJZ , the trend of average growth rate of mycelium was stationary , the rate of transforming total polysaccharides and total triterpenoid were stable ,to a certain ex-tent,it can improve the state of the resource -poor of Inonotus obliquus and lays a foundation for the develop-ment of new pharmacy .%目的:通过对桦褐孔菌与玉米须双向固体发酵工艺的研究,优化其培养条件。方法:采用固体发酵方法得到发酵产物,通过监测菌丝体生长速度获得平均生长速率、通过紫外分光光度法进行总多糖及总三萜含量测定,最后运用加权平均法确定最佳培养条件。结果:最佳培养条件为培养温度(28±2)℃,固体物料玉米须与玉米芯的最佳配比为1∶3,培养料干重90g/袋,液体菌种接种量为2ml/袋,静置培养周期为50天。结论:通过双向固体发酵技术,采用不同配比固体培养基得到的桦须菌质( HXJZ),菌丝平均生长速率趋势平稳,总多糖、总三萜生物转化率稳定,能够从一定程度上改

  10. Research methodology for maize gray leaf spot:Ⅰ.sporulation of Cercospora zeae-maydis%玉米灰斑病研究方法:Ⅰ.玉米灰斑病菌孢子的培养

    Institute of Scientific and Technical Information of China (English)

    赵正龙; 吴兴兴; 毛自朝; 何月秋

    2011-01-01

    为了明确玉米灰斑病菌的产孢条件,将玉米灰斑病菌接种到马铃薯蔗糖(PSA),燕麦番茄碳酸钙(OTCA)、玉米叶粉碳酸钙(MLPCA)、燕麦粉(OA)、粗面粉(CWPA)等5种培养基上,于25℃下培养,测定该菌生长速度、产孢时间和产孢量.结果表明,菌丝在上述培养基上生长速度均较慢,每天平均生长0.33 nun左右,其中以在OA和PSA上生长速度相对较快.基于上述结果,构建了玉米灰斑病菌孢子培养的二级分段培养法,即将菌丝块接种于加有10~15粒直径为5 mm玻璃珠的马铃薯蔗糖(PS)液体培养基三角瓶里,在25℃,180 r/min条件下振荡培养15 d,形成菌丝悬液,再取菌丝悬浮液涂布在MLPCA培养基上,避光培养13 d左右,可获得大童供接种的分生孢子.长期光照促进菌丝生长,不利于孢子的形成,而长期黑暗不利于菌丝生长,但有利于孢子的形成.%The pathogen of maize gray leaf spot, Cercospora zeae-maydis, was inoculated on potato sucrose agar (PSA), oat tomato plus CaCO3 agar(OTCA), maize leaf powder plus CaCO3 agar(MLPCA), oatmeal agar(OA) and crude wheat powder agar (CWPA), and incubated at 25 ℃ for measuring the mycelial growth, sporulation time and number. The results showed that the mycelia grew very slow on the media, with an average of 0.33 mm per day. They grew fastest on OA and PSA, slower on CWPA and most slowly on MLPCA. Based on the results,a sporulation method was set up, namely two-step culturing method. The mycelia were inoculated into flask with PS and 10-15 glass balls of 5mm diameter, and shaken at 25 ℃ and 180 r/min for 15 days, to culture the mycelial fragment suspension. Then the suspension was smeared on MLPCA medium and incubated for about 13 days at 25 ℃ without light. By this way, a large number of spores could be produced for inoculation. Long illumination promoted the mycelial growth, but on the contrary, long darkness was conducive to sporulation.

  11. Metabolic priming by a secreted fungal effector.

    Science.gov (United States)

    Djamei, Armin; Schipper, Kerstin; Rabe, Franziska; Ghosh, Anupama; Vincon, Volker; Kahnt, Jörg; Osorio, Sonia; Tohge, Takayuki; Fernie, Alisdair R; Feussner, Ivo; Feussner, Kirstin; Meinicke, Peter; Stierhof, York-Dieter; Schwarz, Heinz; Macek, Boris; Mann, Matthias; Kahmann, Regine

    2011-10-05

    Maize smut caused by the fungus Ustilago maydis is a widespread disease characterized by the development of large plant tumours. U. maydis is a biotrophic pathogen that requires living plant tissue for its development and establishes an intimate interaction zone between fungal hyphae and the plant plasma membrane. U. maydis actively suppresses plant defence responses by secreted protein effectors. Its effector repertoire comprises at least 386 genes mostly encoding proteins of unknown function and expressed exclusively during the biotrophic stage. The U. maydis secretome also contains about 150 proteins with probable roles in fungal nutrition, fungal cell wall modification and host penetration as well as proteins unlikely to act in the fungal-host interface like a chorismate mutase. Chorismate mutases are key enzymes of the shikimate pathway and catalyse the conversion of chorismate to prephenate, the precursor for tyrosine and phenylalanine synthesis. Root-knot nematodes inject a secreted chorismate mutase into plant cells likely to affect development. Here we show that the chorismate mutase Cmu1 secreted by U. maydis is a virulence factor. The enzyme is taken up by plant cells, can spread to neighbouring cells and changes the metabolic status of these cells through metabolic priming. Secreted chorismate mutases are found in many plant-associated microbes and might serve as general tools for host manipulation.

  12. The Transition from a Phytopathogenic Smut Ancestor to an Anamorphic Biocontrol Agent Deciphered by Comparative Whole-Genome Analysis[W][OPEN

    Science.gov (United States)

    Lefebvre, François; Joly, David L.; Labbé, Caroline; Teichmann, Beate; Linning, Rob; Belzile, François; Bakkeren, Guus; Bélanger, Richard R.

    2013-01-01

    Pseudozyma flocculosa is related to the model plant pathogen Ustilago maydis yet is not a phytopathogen but rather a biocontrol agent of powdery mildews; this relationship makes it unique for the study of the evolution of plant pathogenicity factors. The P. flocculosa genome of ∼23 Mb includes 6877 predicted protein coding genes. Genome features, including hallmarks of pathogenicity, are very similar in P. flocculosa and U. maydis, Sporisorium reilianum, and Ustilago hordei. Furthermore, P. flocculosa, a strict anamorph, revealed conserved and seemingly intact mating-type and meiosis loci typical of Ustilaginales. By contrast, we observed the loss of a specific subset of candidate secreted effector proteins reported to influence virulence in U. maydis as the singular divergence that could explain its nonpathogenic nature. These results suggest that P. flocculosa could have once been a virulent smut fungus that lost the specific effectors necessary for host compatibility. Interestingly, the biocontrol agent appears to have acquired genes encoding secreted proteins not found in the compared Ustilaginales, including necrosis-inducing-Phytophthora-protein- and Lysin-motif- containing proteins believed to have direct relevance to its lifestyle. The genome sequence should contribute to new insights into the subtle genetic differences that can lead to drastic changes in fungal pathogen lifestyles. PMID:23800965

  13. The transition from a phytopathogenic smut ancestor to an anamorphic biocontrol agent deciphered by comparative whole-genome analysis.

    Science.gov (United States)

    Lefebvre, François; Joly, David L; Labbé, Caroline; Teichmann, Beate; Linning, Rob; Belzile, François; Bakkeren, Guus; Bélanger, Richard R

    2013-06-01

    Pseudozyma flocculosa is related to the model plant pathogen Ustilago maydis yet is not a phytopathogen but rather a biocontrol agent of powdery mildews; this relationship makes it unique for the study of the evolution of plant pathogenicity factors. The P. flocculosa genome of ~23 Mb includes 6877 predicted protein coding genes. Genome features, including hallmarks of pathogenicity, are very similar in P. flocculosa and U. maydis, Sporisorium reilianum, and Ustilago hordei. Furthermore, P. flocculosa, a strict anamorph, revealed conserved and seemingly intact mating-type and meiosis loci typical of Ustilaginales. By contrast, we observed the loss of a specific subset of candidate secreted effector proteins reported to influence virulence in U. maydis as the singular divergence that could explain its nonpathogenic nature. These results suggest that P. flocculosa could have once been a virulent smut fungus that lost the specific effectors necessary for host compatibility. Interestingly, the biocontrol agent appears to have acquired genes encoding secreted proteins not found in the compared Ustilaginales, including necrosis-inducing-Phytophthora-protein- and Lysin-motif- containing proteins believed to have direct relevance to its lifestyle. The genome sequence should contribute to new insights into the subtle genetic differences that can lead to drastic changes in fungal pathogen lifestyles.

  14. Genome comparison of barley and maize smut fungi reveals targeted loss of RNA silencing components and species-specific presence of transposable elements.

    Science.gov (United States)

    Laurie, John D; Ali, Shawkat; Linning, Rob; Mannhaupt, Gertrud; Wong, Philip; Güldener, Ulrich; Münsterkötter, Martin; Moore, Richard; Kahmann, Regine; Bakkeren, Guus; Schirawski, Jan

    2012-05-01

    Ustilago hordei is a biotrophic parasite of barley (Hordeum vulgare). After seedling infection, the fungus persists in the plant until head emergence when fungal spores develop and are released from sori formed at kernel positions. The 26.1-Mb U. hordei genome contains 7113 protein encoding genes with high synteny to the smaller genomes of the related, maize-infecting smut fungi Ustilago maydis and Sporisorium reilianum but has a larger repeat content that affected genome evolution at important loci, including mating-type and effector loci. The U. hordei genome encodes components involved in RNA interference and heterochromatin formation, normally involved in genome defense, that are lacking in the U. maydis genome due to clean excision events. These excision events were possibly a result of former presence of repetitive DNA and of an efficient homologous recombination system in U. maydis. We found evidence of repeat-induced point mutations in the genome of U. hordei, indicating that smut fungi use different strategies to counteract the deleterious effects of repetitive DNA. The complement of U. hordei effector genes is comparable to the other two smuts but reveals differences in family expansion and clustering. The availability of the genome sequence will facilitate the identification of genes responsible for virulence and evolution of smut fungi on their respective hosts.

  15. Genome Comparison of Barley and Maize Smut Fungi Reveals Targeted Loss of RNA Silencing Components and Species-Specific Presence of Transposable Elements[W

    Science.gov (United States)

    Laurie, John D.; Ali, Shawkat; Linning, Rob; Mannhaupt, Gertrud; Wong, Philip; Güldener, Ulrich; Münsterkötter, Martin; Moore, Richard; Kahmann, Regine; Bakkeren, Guus; Schirawski, Jan

    2012-01-01

    Ustilago hordei is a biotrophic parasite of barley (Hordeum vulgare). After seedling infection, the fungus persists in the plant until head emergence when fungal spores develop and are released from sori formed at kernel positions. The 26.1-Mb U. hordei genome contains 7113 protein encoding genes with high synteny to the smaller genomes of the related, maize-infecting smut fungi Ustilago maydis and Sporisorium reilianum but has a larger repeat content that affected genome evolution at important loci, including mating-type and effector loci. The U. hordei genome encodes components involved in RNA interference and heterochromatin formation, normally involved in genome defense, that are lacking in the U. maydis genome due to clean excision events. These excision events were possibly a result of former presence of repetitive DNA and of an efficient homologous recombination system in U. maydis. We found evidence of repeat-induced point mutations in the genome of U. hordei, indicating that smut fungi use different strategies to counteract the deleterious effects of repetitive DNA. The complement of U. hordei effector genes is comparable to the other two smuts but reveals differences in family expansion and clustering. The availability of the genome sequence will facilitate the identification of genes responsible for virulence and evolution of smut fungi on their respective hosts. PMID:22623492

  16. Regulation of Rad51-Mediated Homologous Recombination by BRCA2, DSS1 and RAD52

    DEFF Research Database (Denmark)

    Rants, Louise Olthaver Juhl

    Homologous recombination (HR) provides a mechanism to restore integrity and maintain stability of the genetic material. HR is a major pathway for repair of DNA double-strand breaks (DSB), recovery of broken replication forks and generation of meiotic crossovers. The defining step in HR...... in governing the activity of Rad51 and to learn how other recombination-associated proteins such as DSS1 and RAD52 contribute to its regulation. We use the yeast-like fungus Ustilago maydis and the avian DT40 cell line as experimental systems since both have a well-conserved BRCA2-based recombinational repair...

  17. The allergenic significance of certain fungi rarely reported as allergens.

    Science.gov (United States)

    Giannini, E H; Northey, W T; Leathers, C R

    1975-12-01

    The allergenic significance of seven different species of fungi was investigated. Included were Chlorophyllum molybdites, Podaxis pistillaris, Stemonitis ferruginea, Lycogala epidendrum, Fuligo septica, Ustilago maydis and Puccinia cynodontis. All of these fungi have wide distribution patterns and aerially disseminated spores but, because of their unique growth characteristics, are usually not reported in atmospheric fungal surveys. Seventy-eight patients were treated for dermal sensitivity to extracts of the organisms after the spores were extracted in 50% glycerinated Coca's solution. The results represent a six-month test period. Forty-four patients, representing 56% of the total number tested, demonstrated dermal reactivity toward one or more of the extracts.

  18. Basidiomycosis: a review of the literature Basidiomicoses: revisão da literatura

    Directory of Open Access Journals (Sweden)

    Carlos da Silva Lacaz

    1996-10-01

    Full Text Available The basidiomycosis, fungal infections provoked by basidiomycetes or agaric fungi have been recorded at growing frequencies in the medical literature, especially after the advent of AIDS in 1991. The basidiospores of these fungi, scattered in the atmosphere and transported by winds or air currents, reach the maxillary sinuses through the nasal route, most of the times causing signs and symptoms of chronic sinusitis. Basidiomycetes have also been isolated from sputum, especially Schizophyllum commune. Lesions of the buccal mucosa, brain abscesses, onychomycosis and endocarditis have been described, with a growing interest in this type of deep mycosis on the part of mycologists and infectologists. The present paper reports descriptions of mycetism as well as infectious processes caused by basidiomycetes, such as Schizophyllum commune, Ustilago maydis (= Ustilago zeae and Coprinus cinereusAs basidiomicoses, infecções fúngicas provocadas por basidiomicetos ou agáricos vêm sendo registradas cada vez com maior frequência na literatura médica, principalmente após o advento da AIDS/SIDA, em 1981. Os basidiosporos desses fungos, espalhados na atmosfera e veiculados através do vento ou de correntes aéreas, atingem por via nasal os seios maxilares, provocando quadros de sinusite crônica, na maioria das vezes. Do escarro também têm sido isolados basidiomicetos, principalmente o Schizophyllum commune. Lesões da mucosa da boca, abscessos cerebrais, onicomicoses e endocardites já foram descritas, aumentando o interesse dos micologistas e infectologistas para este tipo de micose profunda. O presente trabalho assinala, ao lado dos quadros dc micetismo, processos infecciosos provocados por basidiomicetos, a exemplo do Schizophyllum commune, Ustilago maydis (= Ustilago zeae e Coprinus cinereus.

  19. 燕麦散黑穗病防治药剂筛选及其对燕麦幼苗生长的影响%Screening of Fungicides for Control of Ustilago avenae and Effects of Fungicides on the Seedling Growth of Oat

    Institute of Scientific and Technical Information of China (English)

    张玉霞; 王国基; 姚拓; 马文彬

    2015-01-01

    为更好的防治燕麦(Avena sativa)散黑穗病(Ustilago avenae),筛选可行杀菌剂并测定其对病原冬孢子萌发及燕麦幼苗生长的影响,采用水琼脂平板培养冬孢子萌发法和培养皿滤纸发芽法测定5种杀菌剂对散黑穗病菌冬孢子萌发影响以及药剂拌种对燕麦种子发芽、干物质重、根系性状的影响.结果表明:33.5%喹啉铜悬浮剂对燕麦散黑穗病病原菌冬孢子萌发抑制率为100%,且对种子发芽、干物质积累、根系生长具有促进作用;45%咪鲜胺水乳剂对燕麦散黑穗病病原菌冬孢子萌发抑制率为100%,但其拌种的燕麦种子发芽、干物质积累、根系性状均低于对照;25%嘧菌酯悬浮液、40%氟硅唑乳油、25%苯醚甲环唑乳油对燕麦散黑穗病冬孢子萌发均具有较高抑制作用,且对燕麦种子发芽、干物质积累和根系性状均无影响,其施药剂量分别为75 g·hm-2,60 g·hm-2和1000倍液.

  20. The biotechnological use and potential of plant pathogenic smut fungi.

    Science.gov (United States)

    Feldbrügge, Michael; Kellner, Ronny; Schipper, Kerstin

    2013-04-01

    Plant pathogens of the family Ustilaginaceae parasitise mainly on grasses and cause smut disease. Among the best characterised members of this family are the covered smut fungus Ustilago hordei colonising barley and oat as well as the head smut Sporisorium reilianum and the corn smut Ustilago maydis, both infecting maize. Over the past years, U. maydis in particular has matured into a model system for diverse topics like plant-pathogen interaction, cellular transport processes or DNA repair. Consequently, a broad set of genetic, molecular and system biological methods has been established. This set currently serves as a strong foundation to improve existing and establish novel biotechnological applications. Here, we review four promising aspects covering different fields of applied science: (1) synthesis of secondary metabolites produced at fermenter level. (2) Lipases and other hydrolytic enzymes with potential roles in biocatalytic processes. (3) Degradation of ligno-cellulosic plant materials for biomass conversion. (4) Protein expression based on unconventional secretion, a novel approach inspired by basic research on mRNA transport. Thus, plant pathogenic Ustilaginaceae offer a great potential for future biotechnological applications by combining basic research and applied science.

  1. Importance and Status of Sugarcane Smut (Ustilago scitaminea) in ...

    African Journals Online (AJOL)

    Thus, periodic assessment on the .... towards the last ratoon in all the three sugarcane plantations (Table 1). From the surveyed fields of ... Finchaa and the plantation with more than 50 years of smut history i.e. Wonji-Shoa, indicated that the ...

  2. Analysis of the causes for worsening maize common smut and the countermeasures%玉米黑粉病危害加重原因分析及综合治理对策

    Institute of Scientific and Technical Information of China (English)

    王朝阳; 赵荣艳; 王建胜; 白雪莉; 支艳英

    2011-01-01

    根据近年对豫北安阳市玉米黑粉病发生危害情况的调查结果,总结出该病具有发生因品种而异、区域逐年扩大、危害多集中于雌穗、程度逐年加重、局部受害严重的特点;分析其发生危害加重的原因,主要有田间菌源量大、外来菌源传播扩散、缺乏有效防治措施、气候环境条件适宜、玉米品种抗病性较差、相关法律法规未阻止病害随种子传播等多方面;提出了以选用优质抗病品种、减少菌源、加强裁培管理等农业防治为主,以药剂处理种子和茎叶喷雾等化学防治为辅,同时搞好技术服务、完善相关法律法规等综合治理对策.%Hazard characteristics of common smut caused by the fungus Ustilago maydis was summarized and the causes of its outbreaks were analyzed through surveying in Anyang City in recent years. Ustilago maydis damaged female heads of corns. Common smut has become more serious yearly due to large amounts of U. maydis in fields and propagating pathogen. Moreover, climate conditions, disease resistance of corn breeds and lack of correlative legislation worsened the situation. Comprehensive treatment countermeasures were put forward, giving high priority to agricultural control, assisted with chemical control. In addition, technical service and the relative laws are also required.

  3. Solopathogenic strain formation strongly differs among Ustilaginaceae species.

    Science.gov (United States)

    Sabbagh, Seyed Kazem; Diagne-Lèye, Gnagna; Naudan, Mathieu; Roux, Christophe Paul

    2010-04-01

    The pathogenicity of smut fungi is initiated by the fusion of two compatible saprotrophic yeasts that give rise to the formation of dikaryotic pathogenic hyphae. It has been described in the literature that complementation assays of auxotrophic yeasts of Ustilago maydis have allowed the isolation of diploid strains that are solopathogenic, i.e. pathogenic in the absence of mating. The occurrence of such strains from germinating teliospores was not investigated. We evaluated the ability of teliospores to generate solopathogenic strains in three species of smut fungi: Sporisorium reilianum f.sp. zeae, U. maydis and Moesziomyces penicillariae. Using an approach based on the stability of pseudohyphae of solopathogenic strains, we isolated the strain SRZS1 from teliospores of S. reilianum. Microscopic observations and analyses of mating-type alleles showed that SRZS1 is monokaryotic and diploid. Inoculation tests on maize plantlets indicated that SRZS1 is infectious. The same protocol was applied to polyteliosporal isolates from M. penicillariae, U. maydis and S. reilianum of diverse geographic origin. Surprisingly, all strains from teliospores of M. penicillariae were solopathogenic, whereas only few solopathogenic strains were obtained from the other two species. The possible incidence of solopathogenic strain production in the biology of these species is discussed.

  4. Regulation of Rad51-Mediated Homologous Recombination by BRCA2, DSS1 and RAD52

    DEFF Research Database (Denmark)

    Rants, Louise Olthaver Juhl

    in governing the activity of Rad51 and to learn how other recombination-associated proteins such as DSS1 and RAD52 contribute to its regulation. We use the yeast-like fungus Ustilago maydis and the avian DT40 cell line as experimental systems since both have a well-conserved BRCA2-based recombinational repair...... system that resembles the one seen in human. In U. maydis, we show that Brh2, the BRCA2 homologue, and Dss1 colocalize at DNA damage-induced foci, with Dss1 exhibiting a dynamic association with Brh2 foci. Dss1 focus formation is dependent on interaction with full-length Brh2, and the Dss1-Brh2...... interaction is required for resistance to DNA damage. In avian DT40 cells, we show that endogenously tagged DSS1 redistributes into subnuclear foci in response to DNA damaging agents. However, DSS1 rarely colocalizes with BRCA2. Our data also indicate that both U. maydis Dss1 and avian DSS1 are involved...

  5. Mating type loci of Sporisorium reilianum: novel pattern with three a and multiple b specificities.

    Science.gov (United States)

    Schirawski, Jan; Heinze, Bernadette; Wagenknecht, Martin; Kahmann, Regine

    2005-08-01

    Sporisorium reilianum and Ustilago maydis are two closely related smut fungi, which both infect maize but differ fundamentally in their mode of plant invasion and site of symptom development. As a prelude to studying the molecular basis of these differences, we have characterized the mating type loci of S. reilianum. S. reilianum has two unlinked mating type loci, a and b. Genes in both loci and adjacent regions show a high degree of synteny to the corresponding genes of U. maydis. The b locus occurs in at least five alleles and encodes two subunits of a heterodimeric homeodomain transcription factor, while the a locus encodes a pheromone/receptor system. However, in contrast to that of U. maydis, the a locus of S. reilianum exists in three alleles containing two active pheromone genes each. The alleles of the a locus appear to have arisen through recent recombination events within the locus itself. This has created a situation where each pheromone is specific for recognition by only one mating partner.

  6. A Tale of Genome Compartmentalization: The Evolution of Virulence Clusters in Smut Fungi.

    Science.gov (United States)

    Dutheil, Julien Y; Mannhaupt, Gertrud; Schweizer, Gabriel; M K Sieber, Christian; Münsterkötter, Martin; Güldener, Ulrich; Schirawski, Jan; Kahmann, Regine

    2016-02-12

    Smut fungi are plant pathogens mostly parasitizing wild species of grasses as well as domesticated cereal crops. Genome analysis of several smut fungi including Ustilago maydis revealed a singular clustered organization of genes encoding secreted effectors. In U. maydis, many of these clusters have a role in virulence. Reconstructing the evolutionary history of clusters of effector genes is difficult because of their intrinsically fast evolution, which erodes the phylogenetic signal and homology relationships. Here, we describe the use of comparative evolutionary analyses of quality draft assemblies of genomes to study the mechanisms of this evolution. We report the genome sequence of a South African isolate of Sporisorium scitamineum, a smut fungus parasitizing sugar cane with a phylogenetic position intermediate to the two previously sequenced species U. maydis and Sporisorium reilianum. We show that the genome of S. scitamineum contains more and larger gene clusters encoding secreted effectors than any previously described species in this group. We trace back the origin of the clusters and find that their evolution is mainly driven by tandem gene duplication. In addition, transposable elements play a major role in the evolution of the clustered genes. Transposable elements are significantly associated with clusters of genes encoding fast evolving secreted effectors. This suggests that such clusters represent a case of genome compartmentalization that restrains the activity of transposable elements on genes under diversifying selection for which this activity is potentially beneficial, while protecting the rest of the genome from its deleterious effect.

  7. The Corn Smut (‘Huitlacoche’) as a New Platform for Oral Vaccines

    Science.gov (United States)

    Juárez-Montiel, Margarita; Romero-Maldonado, Andrea; Monreal-Escalante, Elizabeth; Becerra-Flora, Alicia; Korban, Schuyler S.; Rosales-Mendoza, Sergio; Jiménez-Bremont, Juan Francisco

    2015-01-01

    The development of new alternative platforms for subunit vaccine production is a priority in the biomedical field. In this study, Ustilago maydis, the causal agent of common corn smut or ‘huitlacoche’has been genetically engineered to assess expression and immunogenicity of the B subunit of the cholera toxin (CTB), a relevant immunomodulatory agent in vaccinology. An oligomeric CTB recombinant protein was expressed in corn smut galls at levels of up to 1.3 mg g-1 dry weight (0.8% of the total soluble protein). Mice orally immunized with ‘huitlacoche’-derived CTB showed significant humoral responses that were well-correlated with protection against challenge with the cholera toxin (CT). These findings demonstrate the feasibility of using edible corn smut as a safe, effective, and low-cost platform for production and delivery of a subunit oral vaccine. The implications of this platform in the area of molecular pharming are discussed. PMID:26207365

  8. Dicty_cDB: Contig-U14071-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available p... 191 3e-47 NRL( 721P ) H-ras p21 protein mutant with gln 61 replaced by leu (... 191 3e-47 AY892487_1( ...191 3e-47 NRL( 621P ) H-ras p21 protein mutant with gln 61 replaced by his (... 191 3e-47 AY886901_1( AY8869...L( 221P ) H-ras p21 protein mutant with asp 38 replaced by glu (... 190 7e-47 J02207_2( J02207 |pid:none) ha...otein p29; Contains: Rec... 189 1e-46 NRL( 1AGP ) H-ras p21 protein mutant with gly 12 replaced by asp (... ...16 |pid:none) Ustilago maydis small G-protein Ra... 189 2e-46 NRL( 421P ) H-ras p21 protein mutant with gly 12 replace

  9. Mathematical model with spatially uniform regulation explains long-range bidirectional transport of early endosomes in fungal hyphae.

    Science.gov (United States)

    Gou, Jia; Edelstein-Keshet, Leah; Allard, Jun

    2014-08-15

    In many cellular contexts, cargo is transported bidirectionally along microtubule bundles by dynein and kinesin-family motors. Upstream factors influence how individual cargoes are locally regulated, as well as how long-range transport is regulated at the whole-cell scale. Although the details of local, single-cargo bidirectional switching have been extensively studied, it remains to be elucidated how this results in cell-scale spatial organization. Here we develop a mathematical model of early endosome transport in Ustilago maydis. We demonstrate that spatiotemporally uniform regulation, with constant transition rates, results in cargo dynamics that is consistent with experimental data, including data from motor mutants. We find that microtubule arrays can be symmetric in plus-end distribution but asymmetric in binding-site distribution in a manner that affects cargo dynamics and that cargo can travel past microtubule ends in microtubule bundles. Our model makes several testable predictions, including secondary features of dynein and cargo distributions.

  10. Synthesis of new 5-((2-(substituted phenoxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)-1,3,4-oxadiazole-2-thiol: A novel class of potential antibacterial and antifungal agents.

    Science.gov (United States)

    Manjunath, G; Bheemaraju, G; Mahesh, M; Venkata Ramana, P

    2015-11-01

    Novel 1,3,4-Oxadiazoles bearing benzimidazole nucleus were designed, synthesized using 2-(2-(substituted phenoxymethyl)-1H-benzo[d]imidazol-1-yl)acetohydrazide and carbon disulfide. These newly synthesized benzimidazolyl oxadiazoles along with benzimidazolyl acetates and benzimidazolyl acetohydrazides were screened for their antibacterial activity against two kinds of strains using the agar disk diffusion method and antifungal activity against Aspergillus niger and Ustilago maydis. The results showed that some of the compounds exhibited moderate activity against both the strains in antibacterial activity and majority of compounds are not active in antifungal activity. The structure-activity relationships were briefly discussed. The studies indicated that compounds of Benzimidazolyl acetohydrazide were the most potent inhibitors compared to the other compounds under investigation. Copyright © 2015 Académie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.

  11. Membrane-Coupled mRNA Trafficking in Fungi.

    Science.gov (United States)

    Haag, Carl; Steuten, Benedikt; Feldbrügge, Michael

    2015-01-01

    Intracellular logistics are essential for delivery of newly synthesized material during polar growth of fungal hyphae. Proteins and lipids are actively transported throughout the cell by motor-dependent movement of small vesicles or larger units such as endosomes and the endoplasmic reticulum. A remarkably tight link is emerging between active membrane trafficking and mRNA transport, a process that determines the precise subcellular localization of translation products within the cell. Here, we report on recent insights into the mechanism and biological role of these intricate cotransport processes in fungal models such as Saccharomyces cerevisiae, Candida albicans, and Ustilago maydis. In the latter, we focus on the new finding of endosomal mRNA transport and its implications for protein targeting, complex assembly, and septin biology.

  12. Efficacy Test of Triazole Fungicide Against Corn Smut and Discussion on the Calculating Method of Control Effect%三唑类杀菌剂对玉米黑粉病效果测试及防效计算方法探讨

    Institute of Scientific and Technical Information of China (English)

    朱玉新; 陈杰新; 买新红; 赵亚茹

    2015-01-01

    通过试验表明:苯醚甲环唑20%水分散粒剂、戊唑醇80%水分散粒剂、丙环唑25%乳油、三唑酮20%乳油对玉米黑粉病[Ustilago maydis (DC.) Corda.]具有理想的防治效果.其中,病瘤体积防效分别为91.06%、89.04%、88.97%和87.19%,病瘤重量防效分别为92.19%、91.79%、90.73%和89.34%.

  13. Active diffusion and microtubule-based transport oppose myosin forces to position organelles in cells

    Science.gov (United States)

    Lin, Congping; Schuster, Martin; Guimaraes, Sofia Cunha; Ashwin, Peter; Schrader, Michael; Metz, Jeremy; Hacker, Christian; Gurr, Sarah Jane; Steinberg, Gero

    2016-06-01

    Even distribution of peroxisomes (POs) and lipid droplets (LDs) is critical to their role in lipid and reactive oxygen species homeostasis. How even distribution is achieved remains elusive, but diffusive motion and directed motility may play a role. Here we show that in the fungus Ustilago maydis ~95% of POs and LDs undergo diffusive motions. These movements require ATP and involve bidirectional early endosome motility, indicating that microtubule-associated membrane trafficking enhances diffusion of organelles. When early endosome transport is abolished, POs and LDs drift slowly towards the growing cell end. This pole-ward drift is facilitated by anterograde delivery of secretory cargo to the cell tip by myosin-5. Modelling reveals that microtubule-based directed transport and active diffusion support distribution, mobility and mixing of POs. In mammalian COS-7 cells, microtubules and F-actin also counteract each other to distribute POs. This highlights the importance of opposing cytoskeletal forces in organelle positioning in eukaryotes.

  14. The Causes and Prevention Measures of Common Smut in Corn%玉米瘤黑粉病重发原因及防治措施

    Institute of Scientific and Technical Information of China (English)

    史志锋; 史惠琴; 高岩

    2009-01-01

    @@ 玉米瘤黑粉病[Ustilago maydis (DC.) Corda.]是甘肃省泾川县玉米的常见病害,多年来一直发病较轻,未造成为害.2009年由于菌源、气候等综合因素影响、玉米瘤黑粉病再度发生,局部地方为害成灾,已对全县玉米生产构成重大威胁.为此,我们对发病情况进行了实地调查,分析严重发病的原因,并提出相应的防治措施.

  15. A Novel Radiation-Resistant Yeast, Filobasidium elegans RRY1

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Harinder; Kim, Ha Ram; Song, Hyun Pa; Lim, Sang Yong; Kim, Dong Ho [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2012-05-15

    The tolerance to ionizing radiation stress is present among different classes and species of organisms. As listed by Rainey et al., ionizing radiation resistant organisms were isolated from a variety of different sources like processed/canned food items, paper industry, soil and water samples. Apart from extensively reported bacteria and Archea group, many fungal species like Aspergillus, Curvularia, Alternaria, Cryptococcus, and Ustilago maydis have been found to be resistant to ionizing radiation. However, different environmental sources are constantly been explored for novel radioresistant organisms, which can help in understanding the molecular mechanism behind these extreme stress responses. On the basis of this, present study was initiated to find novel radiation resistant yeast from sea water source

  16. Synthesis and fungicidal activity of a series of novel aryloxylepidines.

    Science.gov (United States)

    Kirby, N V; Daeuble, J F; Davis, L N; Hannum, A C; Hellwig, K; Lawler, L K; Parker, M H; Pieczko, M E

    2001-09-01

    A series of novel (hetero) aryloxylepidine derivatives was devised as hybrid structures of the phenoxyquinoline and phenethoxyquin(az)oline fungicides. Synthesis of these targets required the development of several new routes to derivatised 4-hydroxymethylquinolines, and subsequent coupling with phenols or haloarenes. The aryloxylepidines generally showed moderate broad-spectrum fungicidal activity across several diseases of cereals. Substitution of the quinoline ring with chlorine at the 7- and/or 5-positions gave molecules with high levels of protectant activity against Erysiphe graminis f sp tritici (powdery mildew of wheat), but this did not improve the level of fungicidal activity against other diseases. In vitro activity against mitochondrial electron transport complex I (MET) derived from Ustilago maydis showed that 8-fluorolepidine analogues were moderately active at this target site, while the more fungicidally active 7- and 5,7-substituted compounds were inactive. This indicates that MET is not the primary target of these highly active powdery mildewicides.

  17. AcEST: DK953131 [AcEST

    Lifescience Database Archive (English)

    Full Text Available TST38A01NGRL0015_P10 673 Adiantum capillus-veneris mRNA. clone: TST38A01NGRL0015_P1...0. 5' end sequence. DK953131 CL479Contig1 Show DK953131 Clone id TST38A01NGRL0015_P10 Library TST38 Length 6...73 Definition Adiantum capillus-veneris mRNA. clone: TST38A01NGRL0015_P10. 5' end sequence. Accession DK953131...leic Acids Res. 25:3389-3402. Query= DK953131|Adiantum capillus-veneris mRNA, clo...spergillus ter... 132 1e-30 sp|Q4P622|MKAR_USTMA 3-ketoacyl-CoA reductase OS=Ustilago maydis... 131 4e-30 sp

  18. Significant characteristics of the new maize hybrid Rubin-7

    Directory of Open Access Journals (Sweden)

    Jeličić Zora

    2003-01-01

    Full Text Available The Rubin-7 maize hybrid belongs to the FAO 700 maturity group. It is characterized by high yield potential for kernels, which was proven during investigations by the Committee for Species. During the three year monitoring period, from 1999 to 2001, the average yield of kernel was 9.412 t/ha which is 5% above the ZP 704 standard, and was highly statistically significant. Resistance to disease was high for Ustilago maydis 0.49, Fusarium spp. 0.13, and Exerohilum turcicum 1.25. Tolerance against Ostrinia nubilalis is 3-33. All of the above parameters and the agreeable phenotype of this hybrid indicate the value of Rubin-7. .

  19. Crop Management Practices in the Humid Hills from Northeastern Brazil between 670-530 Yrs BP: Palynological Evidences from Archaeological Site Evaristo I

    Directory of Open Access Journals (Sweden)

    Aline Gonçalves Freitas

    2015-12-01

    Full Text Available The first cultural traces of ancient pottery towns in the Serra de Baturité are presented. The pollen spectrum of sediments reveals a mosaic of moist mountainous vegetation, xerophytes, annual nitrophilous, hygrophilous and bog plants. Useful pollen recovered from ceramic, such as cassava (Manihot type, sweet potatoes (Ipomoea type, cotton (Gossypium type, palm trees and fruitful (Arecaceae, cf. Astronium and Anacardium type, together with pathogenic microfungi corn, cotton and some tubers (Curvularia type, Alternaria, Puccinia type and cf. Ustilago maydis indicate agricultural and livelihood activities. The coprophilous fungi of humans and other animals (Cercophora type Gelasinospora type and Sordariaceae reflect the time spent by these groups in the archaeological area. The Gelasinospora fungus also shows the use of fire as fuel for agricultural practices and hunting. These data demonstrate the use of ceramics in funerary and domestic contexts.

  20. Sex in smut fungi: Structure, function and evolution of mating-type complexes.

    Science.gov (United States)

    Bakkeren, Guus; Kämper, Jörg; Schirawski, Jan

    2008-08-01

    Smut fungi are basidiomycete plant pathogens that pose a threat to many important cereal crops. In order to be pathogenic on plants, smut fungal cells of compatible mating-type need to fuse. Fusion and pathogenicity are regulated by two loci, a and b, which harbor conserved genes. The functions of the encoded mating-type complexes have been well-studied in the model fungus Ustilago maydis and will be briefly reviewed here. Sequence comparison of the mating-type loci of different smut and related fungi has revealed that these loci differ substantially in structure. These structural differences point to an evolution from tetrapolar to bipolar mating behavior, which might have occurred several independent times during fungal speciation.

  1. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica.

    Science.gov (United States)

    Bailey, A M; Mena, G L; Herrera-Estrella, L

    1991-08-11

    Phytophthora capsici and P.parasitica were transformed to hygromycin B resistance using plasmids pCM54 and pHL1, which contain the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000. Transformation frequencies of over 500 transformants per micrograms of DNA per 1 x 10(6) protoplasts were obtained. Plasmid pCM54 appears to be transmitted in Phytophthora spp. as an extra-chromosomal element through replication, as shown by Southern blot hybridization and by the loss of plasmid methylation. In addition, transformed strains retained their capacity of infecting Serrano pepper seedlings and Mc. Intosh apple fruits, the host plants for P.capsici and P.parasitica, respectively.

  2. Active diffusion and microtubule-based transport oppose myosin forces to position organelles in cells

    Science.gov (United States)

    Lin, Congping; Schuster, Martin; Guimaraes, Sofia Cunha; Ashwin, Peter; Schrader, Michael; Metz, Jeremy; Hacker, Christian; Gurr, Sarah Jane; Steinberg, Gero

    2016-01-01

    Even distribution of peroxisomes (POs) and lipid droplets (LDs) is critical to their role in lipid and reactive oxygen species homeostasis. How even distribution is achieved remains elusive, but diffusive motion and directed motility may play a role. Here we show that in the fungus Ustilago maydis ∼95% of POs and LDs undergo diffusive motions. These movements require ATP and involve bidirectional early endosome motility, indicating that microtubule-associated membrane trafficking enhances diffusion of organelles. When early endosome transport is abolished, POs and LDs drift slowly towards the growing cell end. This pole-ward drift is facilitated by anterograde delivery of secretory cargo to the cell tip by myosin-5. Modelling reveals that microtubule-based directed transport and active diffusion support distribution, mobility and mixing of POs. In mammalian COS-7 cells, microtubules and F-actin also counteract each other to distribute POs. This highlights the importance of opposing cytoskeletal forces in organelle positioning in eukaryotes. PMID:27251117

  3. Chemical Genetics — A Versatile Method to Combine Science and Higher Level Teaching in Molecular Genetics

    Directory of Open Access Journals (Sweden)

    Björn Sandrock

    2012-10-01

    Full Text Available Phosphorylation is a key event in many cellular processes like cell cycle, transformation of environmental signals to transcriptional activation or polar growth. The chemical genetics approach can be used to analyse the effect of highly specific inhibition in vivo and is a promising method to screen for kinase targets. We have used this approach to study the role of the germinal centre kinase Don3 during the cell division in the phytopathogenic fungus Ustilago maydis. Due to the easy determination of the don3 phenotype we have chosen this approach for a genetic course for M.Sc. students and for IMPRS (International Max-Planck research school students. According to the principle of “problem-based learning” the aim of this two-week course is to transfer knowledge about the broad spectrum of kinases to the students and that the students acquire the ability to design their own analog-sensitive kinase of interest. In addition to these training goals, we benefit from these annual courses the synthesis of basic constructs for genetic modification of several kinases in our model system U. maydis.

  4. The general transcriptional repressor Tup1 is required for dimorphism and virulence in a fungal plant pathogen.

    Directory of Open Access Journals (Sweden)

    Alberto Elías-Villalobos

    2011-09-01

    Full Text Available A critical step in the life cycle of many fungal pathogens is the transition between yeast-like growth and the formation of filamentous structures, a process known as dimorphism. This morphological shift, typically triggered by multiple environmental signals, is tightly controlled by complex genetic pathways to ensure successful pathogenic development. In animal pathogenic fungi, one of the best known regulators of dimorphism is the general transcriptional repressor, Tup1. However, the role of Tup1 in fungal dimorphism is completely unknown in plant pathogens. Here we show that Tup1 plays a key role in orchestrating the yeast to hypha transition in the maize pathogen Ustilago maydis. Deletion of the tup1 gene causes a drastic reduction in the mating and filamentation capacity of the fungus, in turn leading to a reduced virulence phenotype. In U. maydis, these processes are controlled by the a and b mating-type loci, whose expression depends on the Prf1 transcription factor. Interestingly, Δtup1 strains show a critical reduction in the expression of prf1 and that of Prf1 target genes at both loci. Moreover, we observed that Tup1 appears to regulate Prf1 activity by controlling the expression of the prf1 transcriptional activators, rop1 and hap2. Additionally, we describe a putative novel prf1 repressor, named Pac2, which seems to be an important target of Tup1 in the control of dimorphism and virulence. Furthermore, we show that Tup1 is required for full pathogenic development since tup1 deletion mutants are unable to complete the sexual cycle. Our findings establish Tup1 as a key factor coordinating dimorphism in the phytopathogen U. maydis and support a conserved role for Tup1 in the control of hypha-specific genes among animal and plant fungal pathogens.

  5. Pheromone-Related Inhibitors of Ustilago hordei Mating and Tilletia tritici Teliospore Germination

    National Research Council Canada - National Science Library

    Kosted, Paula J; Gerhardt, Shirley A; Sherwood, John E

    2002-01-01

    ... germination inhibition with U. hordei or Tilletia tritici, respectively. N-Acetyl-S-farnesylcysteine, which inhibits processing of Ras, and other sulfur-containing compounds such as homocysteine or methionine, were likewise modified and tested...

  6. Metabolic engineering of Ustilago trichophora TZ1 for improved malic acid production

    Directory of Open Access Journals (Sweden)

    Thiemo Zambanini

    2017-06-01

    These results open up a wide range of possibilities for further optimization, especially combinatorial metabolic engineering to increase the flux from pyruvate to malic acid and to reduce by-product formation.

  7. Bean polygalacturonase inhibitor protein-1 (PGIP-1) inhibits polygalacturonases from Stenocarpella maydis

    CSIR Research Space (South Africa)

    Berger, DK

    2000-07-01

    Full Text Available , apple, raspberry, onion and leek [3, 19, 25?27, 31, 41, 42, 47]. Plant PGIPs interact with endopolygalacturonases from fungi, but do not appear to have an eC128ect on those of bacterial or plant origin [13]. Certain breakdown products of the plant cell... PGs from fungi that the plant was not exposed to during evolution. For example, the bean PGIP-2 interacts strongly with a PG from a pathogen of maize, F. moniliforme [34]. The aim of this study was to investigate PG production of another maize pathogen...

  8. A Gene Cluster for Biosynthesis of Mannosylerythritol Lipids Consisted of 4-O-β-D-Mannopyranosyl-(2R,3S-Erythritol as the Sugar Moiety in a Basidiomycetous Yeast Pseudozyma tsukubaensis.

    Directory of Open Access Journals (Sweden)

    Azusa Saika

    Full Text Available Mannosylerythritol lipids (MELs belong to the glycolipid biosurfactants and are produced by various fungi. The basidiomycetous yeast Pseudozyma tsukubaensis produces diastereomer type of MEL-B, which contains 4-O-β-D-mannopyranosyl-(2R,3S-erythritol (R-form as the sugar moiety. In this respect it differs from conventional type of MELs, which contain 4-O-β-D-mannopyranosyl-(2S,3R-erythritol (S-form as the sugar moiety. While the biosynthetic gene cluster for conventional type of MELs has been previously identified in Ustilago maydis and Pseudozyma antarctica, the genetic basis for MEL biosynthesis in P. tsukubaensis is unknown. Here, we identified a gene cluster involved in MEL biosynthesis in P. tsukubaensis. Among these genes, PtEMT1, which encodes erythritol/mannose transferase, had greater than 69% identity with homologs from strains in the genera Ustilago, Melanopsichium, Sporisorium and Pseudozyma. However, phylogenetic analysis placed PtEMT1p in a separate clade from the other proteins. To investigate the function of PtEMT1, we introduced the gene into a P. antarctica mutant strain, ΔPaEMT1, which lacks MEL biosynthesis ability owing to the deletion of PaEMT1. Using NMR spectroscopy, we identified the biosynthetic product as MEL-A with altered sugar conformation. These results indicate that PtEMT1p catalyzes the sugar conformation of MELs. This is the first report of a gene cluster for the biosynthesis of diastereomer type of MEL.

  9. A Gene Cluster for Biosynthesis of Mannosylerythritol Lipids Consisted of 4-O-β-D-Mannopyranosyl-(2R,3S)-Erythritol as the Sugar Moiety in a Basidiomycetous Yeast Pseudozyma tsukubaensis

    Science.gov (United States)

    Saika, Azusa; Koike, Hideaki; Fukuoka, Tokuma; Yamamoto, Shuhei; Kishimoto, Takahide; Morita, Tomotake

    2016-01-01

    Mannosylerythritol lipids (MELs) belong to the glycolipid biosurfactants and are produced by various fungi. The basidiomycetous yeast Pseudozyma tsukubaensis produces diastereomer type of MEL-B, which contains 4-O-β-D-mannopyranosyl-(2R,3S)-erythritol (R-form) as the sugar moiety. In this respect it differs from conventional type of MELs, which contain 4-O-β-D-mannopyranosyl-(2S,3R)-erythritol (S-form) as the sugar moiety. While the biosynthetic gene cluster for conventional type of MELs has been previously identified in Ustilago maydis and Pseudozyma antarctica, the genetic basis for MEL biosynthesis in P. tsukubaensis is unknown. Here, we identified a gene cluster involved in MEL biosynthesis in P. tsukubaensis. Among these genes, PtEMT1, which encodes erythritol/mannose transferase, had greater than 69% identity with homologs from strains in the genera Ustilago, Melanopsichium, Sporisorium and Pseudozyma. However, phylogenetic analysis placed PtEMT1p in a separate clade from the other proteins. To investigate the function of PtEMT1, we introduced the gene into a P. antarctica mutant strain, ΔPaEMT1, which lacks MEL biosynthesis ability owing to the deletion of PaEMT1. Using NMR spectroscopy, we identified the biosynthetic product as MEL-A with altered sugar conformation. These results indicate that PtEMT1p catalyzes the sugar conformation of MELs. This is the first report of a gene cluster for the biosynthesis of diastereomer type of MEL. PMID:27327162

  10. Gene loss rather than gene gain is associated with a host jump from monocots to dicots in the Smut Fungus Melanopsichium pennsylvanicum.

    Science.gov (United States)

    Sharma, Rahul; Mishra, Bagdevi; Runge, Fabian; Thines, Marco

    2014-07-24

    Smut fungi are well-suited to investigate the ecology and evolution of plant pathogens, as they are strictly biotrophic, yet cultivable on media. Here we report the genome sequence of Melanopsichium pennsylvanicum, closely related to Ustilago maydis and other Poaceae-infecting smuts, but parasitic to a dicot plant. To explore the evolutionary patterns resulting from host adaptation after this huge host jump, the genome of Me. pennsylvanicum was sequenced and compared with the genomes of U. maydis, Sporisorium reilianum, and U. hordei. Although all four genomes had a similar completeness in CEGMA (Core Eukaryotic Genes Mapping Approach) analysis, gene absence was highest in Me. pennsylvanicum, and most pronounced in putative secreted proteins, which are often considered as effector candidates. In contrast, the amount of private genes was similar among the species, highlighting that gene loss rather than gene gain is the hallmark of adaptation after the host jump to the dicot host. Our analyses revealed a trend of putative effectors to be next to another putative effector, but the majority of these are not in clusters and thus the focus on pathogenicity clusters might not be appropriate for all smut genomes. Positive selection studies revealed that Me. pennsylvanicum has the highest number and proportion of genes under positive selection. In general, putative effectors showed a higher proportion of positively selected genes than noneffector candidates. The 248 putative secreted effectors found in all four smut genomes might constitute a core set needed for pathogenicity, whereas those 92 that are found in all grass-parasitic smuts but have no ortholog in Me. pennsylvanicum might constitute a set of effectors important for successful colonization of grass hosts.

  11. Inverse pH regulation of plant and fungal sucrose transporters: a mechanism to regulate competition for sucrose at the host/pathogen interface?

    Directory of Open Access Journals (Sweden)

    Kathrin Wippel

    Full Text Available BACKGROUND: Plant sucrose transporter activities were shown to respond to changes in the extracellular pH and redox status, and oxidizing compounds like glutathione (GSSG or H(2O(2 were reported to effect the subcellular targeting of these proteins. We hypothesized that changes in both parameters might be used to modulate the activities of competing sucrose transporters at a plant/pathogen interface. We, therefore, compared the effects of redox-active compounds and of extracellular pH on the sucrose transporters UmSRT1 and ZmSUT1 known to compete for extracellular sucrose in the Ustilago maydis (corn smut/Zea mays (maize pathosystem. METHODOLOGY/PRINCIPAL FINDINGS: We present functional analyses of the U. maydis sucrose transporter UmSRT1 and of the plant sucrose transporters ZmSUT1 and StSUT1 in Saccharomyces cerevisiae or in Xenopus laevis oocytes in the presence of different extracellular pH-values and redox systems, and study the possible effects of these treatments on the subcellular targeting. We observed an inverse regulation of host and pathogen sucrose transporters by changes in the apoplastic pH. Under none of the conditions analyzed, we could confirm the reported effects of redox-active compounds. CONCLUSIONS/SIGNIFICANCE: Our data suggest that changes in the extracellular pH but not of the extracellular redox status might be used to oppositely adjust the transport activities of plant and fungal sucrose transporters at the host/pathogen interface.

  12. Basidiomycete Mating Type Genes and Pheromone Signaling▿

    Science.gov (United States)

    Raudaskoski, Marjatta; Kothe, Erika

    2010-01-01

    The genome sequences of the basidiomycete Agaricomycetes species Coprinopsis cinerea, Laccaria bicolor, Schizophyllum commune, Phanerochaete chrysosporium, and Postia placenta, as well as of Cryptococcus neoformans and Ustilago maydis, are now publicly available. Out of these fungi, C. cinerea, S. commune, and U. maydis, together with the budding yeast Saccharomyces cerevisiae, have been investigated for years genetically and molecularly for signaling in sexual reproduction. The comparison of the structure and organization of mating type genes in fungal genomes reveals an amazing conservation of genes regulating the sexual reproduction throughout the fungal kingdom. In agaricomycetes, two mating type loci, A, coding for homeodomain type transcription factors, and B, encoding a pheromone/receptor system, regulate the four typical mating interactions of tetrapolar species. Evidence for both A and B mating type genes can also be identified in basidiomycetes with bipolar systems, where only two mating interactions are seen. In some of these fungi, the B locus has lost its self/nonself discrimination ability and thus its specificity while retaining the other regulatory functions in development. In silico analyses now also permit the identification of putative components of the pheromone-dependent signaling pathways. Induction of these signaling cascades leads to development of dikaryotic mycelia, fruiting body formation, and meiotic spore production. In pheromone-dependent signaling, the role of heterotrimeric G proteins, components of a mitogen-activated protein kinase (MAPK) cascade, and cyclic AMP-dependent pathways can now be defined. Additionally, the pheromone-dependent signaling through monomeric, small GTPases potentially involved in creating the polarized cytoskeleton for reciprocal nuclear exchange and migration during mating is predicted. PMID:20190072

  13. The inlfuence of potassium to mineral fertilizers on the maize health

    Institute of Scientific and Technical Information of China (English)

    Jan Bocianowski; Piotr Szulc; Anna Tratwal; Kamila Nowosad; Dariusz Piesik

    2016-01-01

    Field experiments (2009–2011) were conducted at the Department of Agronomy at Poznań University of Life Sciences on the ifelds of the Research Institute in Swadzim. We evaluated the health of maize plants of two types, depending on the variations in mineral fertilization. The conducted research recorded the occurrence of pests such as oscinela frit (Oscinela fritL.) and the European corn borer (Pyrausta nubilalis Hbn.). Diseases recorded during the research included two patho-genes:Fusarium (Fusarium ssp.) and corn smut (Ustilago maydis Corda). It was shown that the meteorological conditions during the maize vegetation had a signiifcant inlfuence on the occurrence of pests. Adding potassium to mineral fertilizers increased the maize resistance toFusarium. Cultivation of “stay-green” cultivar shal be considered as an element of in-tegrated maize protection. The occurrence of oscinela frit was correlated with the occurrence ofFusarium as wel as the occurrence of the European corn borer for both examined cultivars.

  14. Queueing induced by bidirectional motor motion near the end of a microtubule

    Science.gov (United States)

    Ashwin, Peter; Lin, Congping; Steinberg, Gero

    2010-11-01

    Recent live observations of motors in long-range microtubule (MT) dependent transport in the fungus Ustilago maydis have reported bidirectional motion of dynein and an accumulation of the motors at the polymerization-active (the plus-end) of the microtubule. Quantitative data derived from in vivo observation of dynein has enabled us to develop an accurate, quantitatively-valid asymmetric simple exclusion process (ASEP) model that describes the coordinated motion of anterograde and retrograde motors sharing a single oriented microtubule. We give approximate expressions for the size and distribution of the accumulation, and discuss queueing properties for motors entering this accumulation. We show for this ASEP model, that the mean accumulation can be modeled as an M/M/∞ queue that is Poisson distributed with mean Farr/pd , where Farr is the flux of motors that arrives at the tip and pd is the rate at which individual motors change direction from anterograde to retrograde motion. Deviations from this can in principle be used to gain information about other processes at work in the accumulation. Furthermore, our work is a significant step toward a mathematical description of the complex interactions of motors in cellular long-range transport of organelles.

  15. 球毛壳菌60S核糖体蛋白L10a基因克隆与特性分析%Cloning and Characterization Analysis of 60S Ribosomal Protein L10a Gene from Chaetomium globosum

    Institute of Scientific and Technical Information of China (English)

    刘志华; 杨谦

    2006-01-01

    用粗糙脉孢菌(Neurospora crassa)XP_322380和赤霉菌(Gibberella zeae)PH-1(EAA76971)的60S核糖体蛋白L10a基因(60S ribosomal protein L10a,RPL10a)蛋白序列对球毛壳菌(Chaetomium globosum)ESTs序列数据库进行tBlastn检索,获得了球毛壳菌RPL10a cDNA序列.cDNA序列长765 bp,开放阅读框654 bp,编码217个氨基酸组成的多肽,蛋白分子量为23.9 kD.BlastP分析表明该基因氨基酸序列与粗糙脉胞菌相似最高为89%;与玉蜀黍黑粉菌(Ustilago maydis)相似性最低为78%.cDNA序列及推测的氨基酸序列在GenBank登录(登录号分别为AY669070,AAT74578).

  16. Proteomic analysis of the metabolic adaptation of the biocontrol agent Pseudozyma flocculosa leading to glycolipid production

    Directory of Open Access Journals (Sweden)

    Bélanger Richard R

    2010-02-01

    Full Text Available Abstract The yeast-like epiphytic fungus Pseudozyma flocculosa is known to antagonize powdery mildew fungi through proliferation on colonies presumably preceded by the release of an antifungal glycolipid (flocculosin. In culture conditions, P. flocculosa can be induced to produce or not flocculosin through manipulation of the culture medium nutrients. In order to characterize and understand the metabolic changes in P. flocculosa linked to glycolipid production, we conducted a 2-DE proteomic analysis and compared the proteomic profile of P. flocculosa growing under conditions favoring the development of the fungus (control or conducive to flocculosin synthesis (stress. A large number of protein spots (771 were detected in protein extracts of the control treatment compared to only 435 matched protein spots in extracts of the stress cultures, which clearly suggests an important metabolic reorganization in slow-growing cells producing flocculosin. From the latter treatment, we were able to identify 21 protein spots that were either specific to the treatment or up-regulated significantly (2-fold increase. All of them were identified based on similarity between predicted ORF of the newly sequenced genome of P. flocculosa with Ustilago maydis' available annotated sequences. These proteins were associated with the carbon and fatty acid metabolism, and also with the filamentous change of the fungus leading to flocculosin production. This first look into the proteome of P. flocculosa suggests that flocculosin synthesis is elicited in response to specific stress or limiting conditions.

  17. Aspects of DNA repair and nucleotide pool imbalance

    Energy Technology Data Exchange (ETDEWEB)

    Holliday, R.

    1985-01-01

    Evidence that optimum repair depends on adequate pools of deoxynucleotide triphosphates (dNTPs) comes from the study of pyrimidine auxotrophs of Ustilago maydis. These strains are sensitive to UV light and X-rays, and for pyr1-1 it has been shown that the intracellular concentration of dTTP is reduced about 7-fold. The survival curve of pyr1-1 after UV-treatment, and split dose experiments with wild-type cells, provide evidence for an inducible repair mechanism, which probably depends on genetic recombination. Although inducible repair saves cellular resources, it has the disadvantage of becoming ineffective at doses which are high enough to inactivate the repressed structural gene(s) for repair enzymes. It is clear that a wide variety of repair mechanisms have evolved to remove lesions which arise either spontaneously or as a result of damage from external agents. Nevertheless, it would be incorrect to assume that all species require all possible pathways of repair. It is now well established that the accuracy of DNA and protein synthesis depends on proof-reading or editing mechanisms. Optimum accuracy levels will evolve from the balance between error avoidance in macromolecular synthesis and physiological efficiency in growth and propagation.

  18. Molecular cloning, characterization and differential expression of a Sporothrix schenckii STE20-like protein kinase SsSte20.

    Science.gov (United States)

    Zhang, Zhenying; Hou, Binbin; Zheng, Fangliang; Yu, Xiaohong; Liu, Xiaoming

    2013-06-01

    Dimorphic switching requires fungal cells to undergo changes in polarized growth in response to environmental stimuli. The Ste20-related kinases are involved in signaling through mitogen-activated protein kinase pathways and in morphogenesis through the regulation of cytokinesis and actin-dependent polarized growth. In this report, we isolated and characterized an Ste20 homologue gene, designated SsSte20, from yeast-form Sporothrix schenckii (S. schenckii). The full length SsSte20 cDNA sequence is 2846 bp in size, and contains an open reading frame of 2505 bp encoding 835 amino acids. The predicted molecular mass of SsSte20 is 91.31 kDa with an estimated theoretical isoelectric point of 5.76. SsSte20 kinase domain shows 63% identity with that of Don3, a germinal centre kinase (GCK) from Ustilago maydis. Two exons and one intron are identified within the 2578 bp SsSte20 genomic DNA sequence of S. schenckii. Differential expression of the SsSte20 was demonstrated by real-time RT-PCR. The expression of SsSte20 was much higher in the yeast stage compared with that in the mycelial stage, which indicated that the SsSte20 may be involved in the pathogenesis of the yeast phase of S. schenckii.

  19. An expanded family of fungalysin extracellular metallopeptidases of Coprinopsis cinerea.

    Science.gov (United States)

    Lilly, Walt W; Stajich, Jason E; Pukkila, Patricia J; Wilke, Sarah K; Inoguchi, Noriko; Gathman, Allen C

    2008-03-01

    Proteolytic enzymes, particularly secreted proteases of fungal origin, are among the most important of industrial enzymes, yet the biochemical properties and substrate specificities of these proteins have been difficult to characterize. Genomic sequencing offers a powerful tool to identify potentially novel proteases. The genome of the model basidiomycete Coprinopsis cinereus was found to have an unusually high number of metalloproteases that closely match the M36 peptidase family known as fungalysins. The eight predicted C. cinereus fungalysins divide into two groups upon comparison with fungalysins from other fungi. One member, CcMEP1, is most similar to the single representative fungalysins from the basidiomycetes Phanerochaete chrysosporium, Cryptococcus neoformans, and Ustilago maydis, and to the fungalysin type-protein from Aspergillus fumigatus. The remaining seven C. cinereus predicted fungalysins form a group with similarity to three predicted M36 peptidases of Laccaria bicolor. All eight of the C. cinereus enzymes contain both the signature M36 Pfam domain and the FTP propeptide domain. All contain large propeptides with considerable sequence conservation near a proposed cleavage site. The predicted mature enzymes range in size from 37-46 kDa and have isoelectric points that are mildly acidic to neutral. The proximity of these genes to telomeres and/or to transposable elements may have contributed to the expansion of this gene family in C. cinereus.

  20. SCGPred: A Score-based Method for Gene Structure Prediction by Combining Multiple Sources of Evidence

    Institute of Scientific and Technical Information of China (English)

    Xiao Li; Qingan Ren; Yang Weng; Haoyang Cai; Yunmin Zhu; Yizheng Zhang

    2008-01-01

    Predicting protein-coding genes still remains a significant challenge. Although a variety of computational programs that use commonly machine learning methods have emerged, the accuracy of predictions remains a low level when implementing in large genomic sequences. Moreover, computational gene finding in newly sequenced genomes is especially a difficult task due to the absence of a training set of abundant validated genes. Here we present a new gene-finding program, SCGPred,to improve the accuracy of prediction by combining multiple sources of evidence.SCGPred can perform both supervised method in previously well-studied genomes and unsupervised one in novel genomes. By testing with datasets composed of large DNA sequences from human and a novel genome of Ustilago maydi, SCGPred gains a significant improvement in comparison to the popular ab initio gene predictors. We also demonstrate that SCGPred can significantly improve prediction in novel genomes by combining several foreign gene finders with similarity alignments, which is superior to other unsupervised methods. Therefore, SCGPred can serve as an alternative gene-finding tool for newly sequenced eukaryotic genomes. The program is freely available at http://bio.scu.edu.cn/SCGPred/.

  1. Calcium homeostasis and signaling in fungi and their relevance for pathogenicity of yeasts and filamentous fungi

    Directory of Open Access Journals (Sweden)

    Renata Tisi

    2016-09-01

    Full Text Available Though fungi show peculiarities in the purposes and specific traits of calcium signaling pathways, the general scheme and the most important players are well conserved if compared to higher eukaryotes. This provides a powerful opportunity either to investigate shared features using yeast as a model or to exploit fungal specificities as potential targets for antifungal therapies. The sequenced genomes from yeast Saccharomyces cerevisiae, Schizosaccharomyces pombe and the filamentous fungus Neurospora crassa were already published more than ten years ago. More recently the genome sequences of filamentous fungi of Aspergillus genus, some of which threatening pathogens, and dimorphic fungi Ustilago maydis were published, giving the chance to identify several proteins involved in calcium signaling based on their homology to yeast or mammalian counterparts. Nonetheless, unidentified calcium transporters are still present in these organisms which await to be molecularly characterized. Despite the relative simplicity in yeast calcium machinery and the availability of sophisticated molecular tools, in the last years, a number of new actors have been identified, albeit not yet fully characterized. This review will try to describe the state of the art in calcium channels and calcium signaling knowledge in yeast, with particular attention to the relevance of this knowledge with respect to pathological fungi.

  2. The Vip1 inositol polyphosphate kinase family regulates polarized growth and modulates the microtubule cytoskeleton in fungi.

    Directory of Open Access Journals (Sweden)

    Jennifer Pöhlmann

    2014-09-01

    Full Text Available Microtubules (MTs are pivotal for numerous eukaryotic processes ranging from cellular morphogenesis, chromosome segregation to intracellular transport. Execution of these tasks requires intricate regulation of MT dynamics. Here, we identify a new regulator of the Schizosaccharomyces pombe MT cytoskeleton: Asp1, a member of the highly conserved Vip1 inositol polyphosphate kinase family. Inositol pyrophosphates generated by Asp1 modulate MT dynamic parameters independent of the central +TIP EB1 and in a dose-dependent and cellular-context-dependent manner. Importantly, our analysis of the in vitro kinase activities of various S. pombe Asp1 variants demonstrated that the C-terminal phosphatase-like domain of the dual domain Vip1 protein negatively affects the inositol pyrophosphate output of the N-terminal kinase domain. These data suggest that the former domain has phosphatase activity. Remarkably, Vip1 regulation of the MT cytoskeleton is a conserved feature, as Vip1-like proteins of the filamentous ascomycete Aspergillus nidulans and the distantly related pathogenic basidiomycete Ustilago maydis also affect the MT cytoskeleton in these organisms. Consistent with the role of interphase MTs in growth zone selection/maintenance, all 3 fungal systems show aspects of aberrant cell morphogenesis. Thus, for the first time we have identified a conserved biological process for inositol pyrophosphates.

  3. The Vip1 inositol polyphosphate kinase family regulates polarized growth and modulates the microtubule cytoskeleton in fungi.

    Science.gov (United States)

    Pöhlmann, Jennifer; Risse, Carmen; Seidel, Constanze; Pohlmann, Thomas; Jakopec, Visnja; Walla, Eva; Ramrath, Pascal; Takeshita, Norio; Baumann, Sebastian; Feldbrügge, Michael; Fischer, Reinhard; Fleig, Ursula

    2014-09-01

    Microtubules (MTs) are pivotal for numerous eukaryotic processes ranging from cellular morphogenesis, chromosome segregation to intracellular transport. Execution of these tasks requires intricate regulation of MT dynamics. Here, we identify a new regulator of the Schizosaccharomyces pombe MT cytoskeleton: Asp1, a member of the highly conserved Vip1 inositol polyphosphate kinase family. Inositol pyrophosphates generated by Asp1 modulate MT dynamic parameters independent of the central +TIP EB1 and in a dose-dependent and cellular-context-dependent manner. Importantly, our analysis of the in vitro kinase activities of various S. pombe Asp1 variants demonstrated that the C-terminal phosphatase-like domain of the dual domain Vip1 protein negatively affects the inositol pyrophosphate output of the N-terminal kinase domain. These data suggest that the former domain has phosphatase activity. Remarkably, Vip1 regulation of the MT cytoskeleton is a conserved feature, as Vip1-like proteins of the filamentous ascomycete Aspergillus nidulans and the distantly related pathogenic basidiomycete Ustilago maydis also affect the MT cytoskeleton in these organisms. Consistent with the role of interphase MTs in growth zone selection/maintenance, all 3 fungal systems show aspects of aberrant cell morphogenesis. Thus, for the first time we have identified a conserved biological process for inositol pyrophosphates.

  4. [Pityriasis versicolor : new aspects of an old disease].

    Science.gov (United States)

    Mayser, P A; Preuss, J

    2012-11-01

    Pityriasis versicolor (PV) is one of the most common infectious skin diseases, as well as the most common dermatosis associated with pigmentation alterations of the skin. PV is prevalent in 1% of the population living in temperate climate zones and more common during the summer. In tropical areas, PV is found in up to 50% of all patients consulting a dermatologist. Of the known Malassezia species, M. globosa is currently felt to play a key role in the pathogenesis of PV, as it is most commonly found in PV lesions. In addition, its round-shaped cells may contribute to the characteristic histology of the disease ("spaghetti and meatballs"). However, the clinical appearance of PV including hyper- and hypopigmentation, fluorescence of the lesions, as well as a lack of inflammation despite high fungal load cannot fully be explained by the presence of M. globosa, which is also found on healthy skin. In M. furfur a tryptophan-dependent metabolic pathway generates a number of indole pigments, which may be associated with the clinical appearance of PV. In the model organism Ustilago maydis it was shown that the formation of the indole compounds occurs spontaneously after initial conversion of tryptophan into indole pyruvate controlled by the key enzyme aminotransferase Tam 1. We review the present knowledge of PV and highlight the potential role of Tam1 in explaining the poorly understood aspects of the disease. Promising therapeutic results using the application of Tam1 inhibitors to treat PV support the enzyme's important role in the disease pathogenesis.

  5. Lowering intercellular melatonin levels by transgenic analysis of indoleamine 2,3-dioxygenase from rice in tomato plants.

    Science.gov (United States)

    Okazaki, Masateru; Higuchi, Kenji; Aouini, Asma; Ezura, Hiroshi

    2010-10-01

    Melatonin exists in numerous living organisms including vertebrates, insects, fungi, bacteria, and plants. Extensive studies have been conducted on the physiological roles of melatonin in various plant species. In plants, melatonin seems to act in antioxidant protection, as a growth promoter, and in photoperiodism. However, the mechanisms by which melatonin carries out these roles remain unclear. We manipulated the endogenous melatonin content in tomato plants by modifying the metabolic enzyme indoleamine 2,3-dioxygenase (IDO). The OsIDO gene was isolated from rice (Oryza sativa) and characterized using 3-D homology modeling and reverse genetic approaches. The amino acid sequence of OsIDO showed high homology to the Ustilago maydis IDO. The 3-D model structure of OsIDO is composed of a small and a large domain. Transgenic tomato plants constitutively expressing the OsIDO gene exhibited a decrease in their melatonin content. Moreover, the number of lateral leaflets decreased in transgenic plants. Protein extracts taken from these plants showed activity degradation, demonstrating the function of OsIDO. These results suggest the involvement of IDO in plant melatonin metabolism and a possible role in plant leaf development. © 2010 The Authors. Journal of Pineal Research © 2010 John Wiley & Sons A/S.

  6. Występowanie głowni smugowej traw (Ustilago striiformis (Westend. Niessl na rajgrasie wyniosłym w Polsce [The occurrence of the stripe smut of grasses (Ustilago stritformis (Westend. Niessl on Arrhenatherum elatius in Poland

    Directory of Open Access Journals (Sweden)

    J. W. Tomala-Bednarek

    2015-06-01

    Full Text Available The stripe smut of grasses (Ustalago striiformis rarely occurs on Arrhenatherum elatius. It has been collected on this species of grass for the first time in Poland on July 1, 1959, by the author. It has been found that it occurs on tall oat grass from May until late fall. It attacks all aboveground parts of tall oat grass. Strongly smutted plants die off, while the slightly smutted ones may recover.

  7. Sex-specific costs of resistance to the fungal pathogen Ustilago violacea (Microbotryum violaceum) in Silene alba

    NARCIS (Netherlands)

    Biere, A.; Antonovics, J.

    1996-01-01

    Costs of resistance are often invoked to explain the maintenance of polymorphisms for resistance to fungal pathogens in natural plant populations. To investigate such costs, 27 half-sib families of Silene alba, collected from a single host population, were grown in experiment populations in the pres

  8. Preliminary Trial of 11 New Hybrid Maize Genotype to The Resistance on Java Downy Mildew (Peronosclerospora maydis)

    OpenAIRE

    Budi Setyawan; Irfan Suliansyah; Aswaldi Anwar; Etti Swasti

    2016-01-01

    Maize or corn (Zea mays L.) belongs to the family of grasses (Poaceae).  Maize is grown globally and one of the most important cereal crop in the world.  In many countries, corn is the main agricultural crop, and are used as food, feed and industrial raw materials. Together with rice and wheat, corn included in the cereals that provide about 65% carbohydrates and 50% protein that humans need. For this purpose, many developing countries, especially in Asia and Africa are in a strong effort to ...

  9. Enhanced Survival and Nodule Occupancy of Pigeon pea Nodulating Rhizobium sp. ST1 expressing fegA Gene of Bradyrhizobium japonicum 61A152

    Directory of Open Access Journals (Sweden)

    G. Archana

    2009-01-01

    Full Text Available Problem statement: Rhizobial isolates belonging to genera (Rhizobium sp. and Mesorhizobium sp. in our laboratory produced only catecholate type of siderophores. Although FhuA and FegA (ferrichrome receptors homologs were found to be present in the sequenced genomes of few rhizobia (e.g., 1 in R. etli and 2 in Mesorhizobium sp. BNC1, laboratory isolates of the corresponding genera failed to utilize ferrichrome, a siderophore which is present in nanomolar concentrations in the soil. This inability was considered as a negative fitness factor with respect to rhizospheric colonization by these rhizobia. Approach: The 2.4 kb fegA gene (encoding ferrichrome receptor was amplified along with its native promoter from Bradyrhizobium japonicum 61A152 and cloned in a broad host range plasmid vector pUCPM18. The plasmid construct pFJ was transferred by conjugation into Rhizobium sp. ST1 to give transconjugant ST1pFJ12. The consequence of FegA expression on the transconjugant was tested under lab and soil conditions, using physiological experiments. Results: Ability of the transconjugant ST1pFJ12 to utilize ferrichrome and expression of a 79 kD protein band on the outer membrane of the transconjugant confirmed FegA expression. Transconjugant ST1pFJ12 exhibited increased growth rate as compared to the parent strain ST1, in minimal media containing ferrichrome as the sole iron source, confirming the positive effect of FegA expression. Inoculation of pigeon pea seedlings with transconjugant ST1pFJ12 led to a marked increase in plant growth parameters as compared to plants inoculated with the parent strain ST1, the effect being more pronounced when Ustilago maydis, a ferrichrome producer was co-inoculated in the systems. Nodule occupancy on pigeon pea plant when inoculated with the transconjugant ST1pFJ12 alone was 57% which increased to 66% when co-inoculated with U. maydis as compared with 37 and 30

  10. Incidence and distribution of seed-borne fungi associated with wheat in Markazi Province, Iran

    National Research Council Canada - National Science Library

    Masomeh Hajihasani

    2012-01-01

    ... including Tilletia laevis, Tilletia tritici, Ustilago tritici, Fusarium graminearum, Fusarium culmorum, Microdochium nivale, Bipolaris sorokiniana, Alternaria alternata, Curvularia sp., Aspergil...

  11. Identification of growth stage molecular markers in Trichoderma sp. 'atroviride type B' and their potential application in monitoring fungal growth and development in soil.

    Science.gov (United States)

    Mendoza-Mendoza, Artemio; Steyaert, Johanna; Nieto-Jacobo, Maria Fernanda; Holyoake, Andrew; Braithwaite, Mark; Stewart, Alison

    2015-11-01

    Several members of the genus Trichoderma are biocontrol agents of soil-borne fungal plant pathogens. The effectiveness of biocontrol agents depends heavily on how they perform in the complex field environment. Therefore, the ability to monitor and track Trichoderma within the environment is essential to understanding biocontrol efficacy. The objectives of this work were to: (a) identify key genes involved in Trichoderma sp. 'atroviride type B' morphogenesis; (b) develop a robust RNA isolation method from soil; and (c) develop molecular marker assays for characterizing morphogenesis whilst in the soil environment. Four cDNA libraries corresponding to conidia, germination, vegetative growth and conidiogenesis were created, and the genes identified by sequencing. Stage specificity of the different genes was confirmed by either Northern blot or quantitative reverse-transcriptase PCR (qRT-PCR) analysis using RNA from the four stages. con10, a conidial-specific gene, was observed in conidia, as well as one gene also involved in subsequent stages of germination (L-lactate/malate dehydrogenase encoding gene). The germination stage revealed high expression rates of genes involved in amino acid and protein biosynthesis, while in the vegetative-growth stage, genes involved in differentiation, including the mitogen-activated protein kinase kinase similar to Kpp7 from Ustilago maydis and the orthologue to stuA from Aspergillus nidulans, were preferentially expressed. Genes involved in cell-wall synthesis were expressed during conidiogenesis. We standardized total RNA isolation from Trichoderma sp. 'atroviride type B' growing in soil and then examined the expression profiles of selected genes using qRT-PCR. The results suggested that the relative expression patterns were cyclic and not accumulative.

  12. Survey and analysis of simple sequence repeats in the Laccaria bicolor genome, with development of microsatellite markers

    Energy Technology Data Exchange (ETDEWEB)

    Labbe, Jessy L [ORNL; Murat, Claude [INRA, Nancy, France; Morin, Emmanuelle [INRA, Nancy, France; Le Tacon, F [UMR, France; Martin, Francis [INRA, Nancy, France

    2011-01-01

    It is becoming clear that simple sequence repeats (SSRs) play a significant role in fungal genome organization, and they are a large source of genetic markers for population genetics and meiotic maps. We identified SSRs in the Laccaria bicolor genome by in silico survey and analyzed their distribution in the different genomic regions. We also compared the abundance and distribution of SSRs in L. bicolor with those of the following fungal genomes: Phanerochaete chrysosporium, Coprinopsis cinerea, Ustilago maydis, Cryptococcus neoformans, Aspergillus nidulans, Magnaporthe grisea, Neurospora crassa and Saccharomyces cerevisiae. Using the MISA computer program, we detected 277,062 SSRs in the L. bicolor genome representing 8% of the assembled genomic sequence. Among the analyzed basidiomycetes, L. bicolor exhibited the highest SSR density although no correlation between relative abundance and the genome sizes was observed. In most genomes the short motifs (mono- to trinucleotides) were more abundant than the longer repeated SSRs. Generally, in each organism, the occurrence, relative abundance, and relative density of SSRs decreased as the repeat unit increased. Furthermore, each organism had its own common and longest SSRs. In the L. bicolor genome, most of the SSRs were located in intergenic regions (73.3%) and the highest SSR density was observed in transposable elements (TEs; 6,706 SSRs/Mb). However, 81% of the protein-coding genes contained SSRs in their exons, suggesting that SSR polymorphism may alter gene phenotypes. Within a L. bicolor offspring, sequence polymorphism of 78 SSRs was mainly detected in non-TE intergenic regions. Unlike previously developed microsatellite markers, these new ones are spread throughout the genome; these markers could have immediate applications in population genetics.

  13. Domestication of maize, sorghum, and sugarcane did not drive the divergence of their smut pathogens.

    Science.gov (United States)

    Munkacsi, Andrew B; Stoxen, Sam; May, Georgiana

    2007-02-01

    We investigated two alternative hypotheses for the origin of crop pathogen species: that human-mediated agricultural practices drove the divergence of many crop plant pathogen species or that coevolutionary processes in natural populations of the crops' ancestors drove divergence of pathogen species. We distinguished between these two hypotheses by constructing a robust multigene phylogeny and estimating the dates of divergence among four, monophyletic species of smut fungi (Ustilago maydis, U. scitaminea, Sporisorium reilianum, S. sorghi) known to specifically infect maize, sorghum, sugarcane, and their wild ancestors. Without a fossil record for smut fungi, we calibrated the pathogen species' divergence times to their plant host divergence times. Specifically, a calibration date of 10,000 years was employed to test the hypothesis that the fungal species originated at the time of domestication of their current hosts and a calibration date of 50 million years was employed to test the hypothesis that the fungal species originated on wild ancestors of their domesticated hosts. Substitution rates at five protein coding genes were calculated and rates obtained for the 10,000 year calibration date were orders of magnitude faster than those commonly reported for eukaryotes, thus rejecting the hypothesis that these smut pathogen species diverged at the time of domestication. In contrast, substitution rates obtained for the 50 million year calibration were comparable to eukaryotic substitution rates. We used the 50 million year calibration to estimate divergence times of taxa in two datasets, one comprised solely the focal species and one comprised the focal species and additional related taxa. Both datasets indicate that all taxa diverged millions of years ago, strongly supporting the hypothesis that smut species diverged before the time of domestication and modern agriculture. Thus, smut species diverged in the ecological context of natural host plant and fungal

  14. On the origin of 3-methylglutaconic acid in disorders of mitochondrial energy metabolism.

    Science.gov (United States)

    Ikon, Nikita; Ryan, Robert O

    2016-09-01

    3-methylglutaconic acid (3MGA)-uria occurs in numerous inborn errors of metabolism (IEM) associated with compromised mitochondrial energy metabolism. This organic acid arises from thioester cleavage of 3-methylglutaconyl CoA (3MG CoA), an intermediate in leucine catabolism. In individuals harboring mutations in 3MG CoA hydratase (i.e., primary 3MGA-uria), dietary leucine is the source of 3MGA. In secondary 3MGA-uria, however, no leucine metabolism defects have been reported. While others have suggested 3MGA arises from aberrant isoprenoid shunting from cytosol to mitochondria, an alternative route posits that 3MG CoA arises in three steps from mitochondrial acetyl CoA. Support for this biosynthetic route in IEMs is seen by its regulated occurrence in microorganisms. The fungus, Ustilago maydis, the myxobacterium, Myxococcus xanthus and the marine cyanobacterium, Lyngbya majuscule, generate 3MG CoA (or acyl carrier protein derivative) in the biosynthesis of iron chelating siderophores, iso-odd chain fatty acids and polyketide/nonribosomal peptide products, respectively. The existence of this biosynthetic machinery in these organisms supports a model wherein, under conditions of mitochondrial dysfunction, accumulation of acetyl CoA in the inner mitochondrial space as a result of inefficient fuel utilization drives de novo synthesis of 3MG CoA. Since humans lack the downstream biosynthetic capability of the organisms mentioned above, as 3MG CoA levels rise, thioester hydrolysis yields 3MGA, which is excreted in urine as unspent fuel. Understanding the metabolic origins of 3MGA may increase its utility as a biomarker.

  15. Motor-mediated bidirectional transport along an antipolar microtubule bundle: a mathematical model.

    Science.gov (United States)

    Lin, Congping; Ashwin, Peter; Steinberg, Gero

    2013-05-01

    Long-distance bidirectional transport of organelles depends on the coordinated motion of various motor proteins on the cytoskeleton. Recent quantitative live cell imaging in the elongated hyphal cells of Ustilago maydis has demonstrated that long-range motility of motors and their endosomal cargo occurs on unipolar microtubules (MTs) near the extremities of the cell. These MTs are bundled into antipolar bundles within the central part of the cell. Dynein and kinesin-3 motors coordinate their activity to move early endosomes (EEs) in a bidirectional fashion where dynein drives motility towards MT minus ends and kinesin towards MT plus ends. Although this means that one can easily assign the drivers of bidirectional motion in the unipolar section, the bipolar orientations in the bundle mean that it is possible for either motor to drive motion in either direction. In this paper we use a multilane asymmetric simple exclusion process modeling approach to simulate and investigate phases of bidirectional motility in a minimal model of an antipolar MT bundle. In our model, EE cargos (particles) change direction on each MT with a turning rate Ω and there is switching between MTs in the bundle at the minus ends. At these ends, particles can hop between MTs with rate q(1) on passing from a unipolar to a bipolar section (the obstacle-induced switching rate) or q(2) on passing in the other direction (the end-induced switching rate). By a combination of numerical simulations and mean-field approximations, we investigate the distribution of particles along the MTs for different values of these parameters and of Θ, the overall density of particles within this closed system. We find that even if Θ is low, the system can exhibit a variety of phases with shocks in the density profiles near plus and minus ends caused by queuing of particles. We discuss how the parameters influence the type of particle that dominates active transport in the bundle.

  16. MODEL ORGANISMS USED IN MOLECULAR BIOLOGY OR MEDICAL RESEARCH

    Directory of Open Access Journals (Sweden)

    Pandey Govind

    2011-11-01

    Full Text Available A model organism is a non-human species that is studied to understand specific biological phenomena with the expectation that investigations made in the organism model will provide insight into the workings of other organisms. The model organisms are widely used to explore potential causes and treatments for human as well as animal diseases when experiments on animals or humans would be unfeasible or considered less ethical. Studying model organisms may be informative, but care must be taken when generalizing from one organism to another. Often, model organisms are chosen on the basis that they are amenable to experimental manipulation. When researchers look for an organism to use in their studies, they look for several traits. Among these are size, generation time, accessibility, manipulation, genetics, conservation of mechanisms and potential economic benefit. As comparative molecular biology has become more common, some researchers have sought model organisms from a wider assortment of lineages on the tree of life. There are many model organisms, such as viruses (e.g., Phage lambda virus, Tobacco mosaic virus, etc., bacteria (e.g., Bacillus subtilis, Escherichia coli, Pseudomonas fluorescens, Vibrio fischeri, etc., algae (e.g., Chlamydomonas reinhardtii, Emiliania huxleyi, etc., molds (e.g., Aspergillus nidulans, Neurospora crassa, etc., yeasts (e.g., Saccharomyces cerevisiae, Ustilago maydis, etc., higher plants (e.g., Arabidopsis thaliana, Lemna gibba, Lotus japonicus, Nicotiana tabaccum, Oryza sativa, Physcomitrella patens, Zea mays, etc. and animals (e.g., Caenorhabditis elegans, guinea pig, hamster, mouse, rat, cat, chicken, dog, frog, Hydra, Drosophila melanogaster fruit fly, fish, etc..

  17. Gene Network Polymorphism Illuminates Loss and Retention of Novel RNAi Silencing Components in the Cryptococcus Pathogenic Species Complex.

    Directory of Open Access Journals (Sweden)

    Marianna Feretzaki

    2016-03-01

    Full Text Available RNAi is a ubiquitous pathway that serves central functions throughout eukaryotes, including maintenance of genome stability and repression of transposon expression and movement. However, a number of organisms have lost their RNAi pathways, including the model yeast Saccharomyces cerevisiae, the maize pathogen Ustilago maydis, the human pathogen Cryptococcus deuterogattii, and some human parasite pathogens, suggesting there may be adaptive benefits associated with both retention and loss of RNAi. By comparing the RNAi-deficient genome of the Pacific Northwest Outbreak C. deuterogattii strain R265 with the RNAi-proficient genomes of the Cryptococcus pathogenic species complex, we identified a set of conserved genes that were lost in R265 and all other C. deuterogattii isolates examined. Genetic and molecular analyses reveal several of these lost genes play roles in RNAi pathways. Four novel components were examined further. Znf3 (a zinc finger protein and Qip1 (a homolog of N. crassa Qip were found to be essential for RNAi, while Cpr2 (a constitutive pheromone receptor and Fzc28 (a transcription factor are involved in sex-induced but not mitosis-induced silencing. Our results demonstrate that the mitotic and sex-induced RNAi pathways rely on the same core components, but sex-induced silencing may be a more specific, highly induced variant that involves additional specialized or regulatory components. Our studies further illustrate how gene network polymorphisms involving known components of key cellular pathways can inform identification of novel elements and suggest that RNAi loss may have been a core event in the speciation of C. deuterogattii and possibly contributed to its pathogenic trajectory.

  18. Analysis of simple sequence repeats in genomes of Sclerotinia sclerotiorum and Botrytis cinerea%核盘菌和灰葡萄孢基因组中的简单重复序列分析

    Institute of Scientific and Technical Information of China (English)

    陈怀谷; 李伟; 张爱香; 陈丽华; 姜伟丽

    2007-01-01

    利用公共的真菌基因组数据库资源,对核盘菌(Sclerotinia sclerotiorum)和灰葡萄孢(Botrytis cinerea)基因组中SSRs的结构类型、分布、丰度及最长序列等进行了系统分析,并与已经研究过的禾谷镰孢菌(Fusarium graminearum),稻瘟病菌(Magnaporthe grisea)和黑粉菌(Ustilago maydis)等几种植物病原真菌基因组中的SSRs进行了比较.结果表明:核盘菌和灰葡萄孢基因组中的SSRs非常丰富,分别为6 539和8 627个,并且在结构类型和分布规律上具有一定的相似性;与其他几种病原真菌相比,核盘菌和灰葡萄孢基因组中长重复的四、五、六核苷酸基序更为丰富,从而使得这两种真菌具有更高的变异性.同时,我们发现真菌基因组中SSRs的丰度与基因组的大小及GC含量没有必然的关系.文章对核盘菌和灰葡萄孢基因组中SSRs的丰度、出现频率及最长基序的分析为快速、便捷地设计多态性丰富的SSRs引物提供了有益的信息.

  19. The mating type locus (MAT and sexual reproduction of Cryptococcus heveanensis: insights into the evolution of sex and sex-determining chromosomal regions in fungi.

    Directory of Open Access Journals (Sweden)

    Banu Metin

    2010-05-01

    Full Text Available Mating in basidiomycetous fungi is often controlled by two unlinked, multiallelic loci encoding homeodomain transcription factors or pheromones/pheromone receptors. In contrast to this tetrapolar organization, Cryptococcus neoformans/Cryptococcus gattii have a bipolar mating system, and a single biallelic locus governs sexual reproduction. The C. neoformans MAT locus is unusually large (>100 kb, contains >20 genes, and enhances virulence. Previous comparative genomic studies provided insights into how this unusual MAT locus might have evolved involving gene acquisitions into two unlinked loci and fusion into one contiguous locus, converting an ancestral tetrapolar system to a bipolar one. Here we tested this model by studying Cryptococcus heveanensis, a sister species to the pathogenic Cryptococcus species complex. An extant sexual cycle was discovered; co-incubating fertile isolates results in the teleomorph (Kwoniella heveanensis with dikaryotic hyphae, clamp connections, septate basidia, and basidiospores. To characterize the C. heveanensis MAT locus, a fosmid library was screened with C. neoformans/C. gattii MAT genes. Positive fosmids were sequenced and assembled to generate two large probably unlinked MAT gene clusters: one corresponding to the homeodomain locus and the other to the pheromone/receptor locus. Strikingly, two divergent homeodomain genes (SXI1, SXI2 are present, similar to the bE/bW Ustilago maydis paradigm, suggesting one or the other homeodomain gene was recently lost in C. neoformans/C. gattii. Sequencing MAT genes from other C. heveanensis isolates revealed a multiallelic homeodomain locus and at least a biallelic pheromone/receptor locus, similar to known tetrapolar species. Taken together, these studies reveal an extant C. heveanensis sexual cycle, define the structure of its MAT locus consistent with tetrapolar mating, and support the proposed evolutionary model for the bipolar Cryptococcus MAT locus revealing

  20. Common motifs in the response of cereal primary metabolism to fungal pathogens are not based on similar transcriptional reprogramming

    Directory of Open Access Journals (Sweden)

    Lars Matthias Voll

    2011-08-01

    Full Text Available During compatible interactions with their host plants, biotrophic plant pathogens subvert host metabolism to ensure the sustained provision of nutrient assimilates by the colonized host cells. To investigate, whether common motifs can be revealed in the response of primary carbon and nitrogen metabolism towards colonization with biotrophic fungi in cereal leaves, we have conducted a combined metabolome and transcriptome study of three quite divergent pathosystems, the barley powdery mildew fungus (Blumeria graminis f.sp. hordei, the corn smut fungus Ustilago maydis and the maize anthracnose fungus Colletotrichum graminicola, the latter being a hemibiotroph that only exhibits an initial biotrophic phase during its establishment.Based on the analysis of 42 water-soluble metabolites, we were able to separate early biotrophic from late biotrophic interactions by hierarchical cluster analysis and principal component analysis, irrespective of the plant host. Interestingly, the corresponding transcriptome dataset could not discriminate between these stages of biotrophy, irrespective, of whether transcript data for genes of central metabolism or the entire transcriptome dataset was used. Strong differences in the transcriptional regulation of photosynthesis, glycolysis, the TCA cycle, lipid biosynthesis, and cell wall metabolism were observed between the pathosystems. Increased contents of Gln, Asn, and glucose as well as diminished contents of PEP and 3-PGA were common to early post-penetration stages of all interactions. On the transcriptional level, genes of the TCA cycle, nucleotide energy metabolism and amino acid biosynthesis exhibited consistent trends among the compared biotrophic interactions, identifying the requirement for metabolic energy and the rearrangement of amino acid pools as common transcriptional motifs during early biotrophy. Both metabolome and transcript data were employed to generate models of leaf primary metabolism during

  1. Molecular characterization of a bipartite double-stranded RNA virus and its satellite-like RNA co-infecting the phytopathogenic fungus Sclerotinia sclerotiorum

    Directory of Open Access Journals (Sweden)

    Lijiang eLiu

    2015-05-01

    Full Text Available A variety of mycoviruses have been found in Sclerotinia sclerotiorum. In this study, we report a novel mycovirus Sclerotinia sclerotiorum botybirnavirus 1 (SsBRV1 that was originally isolated from the hypovirulent strain SCH941 of S. sclerotiorum. SsBRV1 has rigid spherical virions that are ~38 nm in diameter, and three dsRNA segments (dsRNA1, 2 and 3 with lengths of 6.4, 6.0 and 1.7 kbp, respectively were packaged in the virions. dsRNA1 encodes a cap-pol fusion protein, and dsRNA2 encodes a polyprotein with unknown functions but contributes to the formation of virus particles. The dsRNA3 is dispensable and may be a satellite-like RNA (SatlRNA of SsBRV1. Although phylogenetic analysis of the RdRp domain demonstrated that SsBRV1 is related to Botrytis porri RNA virus 1 (BpRV1 and Ustilago maydis dsRNA virus-H1 (UmV-H1, the structure proteins of SsBRV1 do not have any significant sequence similarities with other known viral proteins with the exception of those of BpRV1. SsBRV1 carrying dsRNA3 seems to have no obvious effects on the colony morphology, but can significantly reduce the growth rate and virulence of S. sclerotiorum. Notably, a growth hormone receptor binding domain (GHBP, Pfam12772 is detected in ORF2-encoded protein of SsBRV1, which have not been reported in any other viruses. These findings provide new insights into the virus taxonomy, virus evolution and the interactions between SsBRV1 and the fungal hosts.

  2. Cloning and Sequence Analysis of a Ser/Thr Protein Kinase Homologous Gene ( vv-stpk 1) from Volvariella volvacea%草菇丝/苏氨酸蛋白激酶同源基因的克隆及序列分析

    Institute of Scientific and Technical Information of China (English)

    汪虹; 陈明杰; 鲍大鹏

    2011-01-01

    根据担子菌丝/苏氨酸蛋白激酶同源基因编码的氨基酸序列设计两对简并引物,通过巢式简并PCR方法获得草菇(Volvariella volvacea)丝/苏氨酸蛋白激酶同源基因(vv-stpk1)的保守片段,然后通过基因组步行的方法获得了vv-stpk1全长序列.vv-stpk1全长为2 003 bp,含有8个内含子,长度分别为72、57、53、54、45、50、55和48 bp,可编码522个氨基酸残基的多肽.推定的氨基酸序列与玉米黑粉菌(Ustilago maydis)、木糖发酵酵母(Pichia stipitis)、新型隐球菌(Cryptococcus neoformans)中与细胞形态发生相关的丝/苏氨酸蛋白激酶Orb6的相似性分别为81%、77%和76%,对VV-STPK1蛋白的系统发生学分析的结果表明,VV-STPK1与Orb6同源蛋白聚在同一进化枝上,这些数据都支持VV-STPK1蛋白为与细胞形态发生相关同源蛋白的推定.

  3. High abundance of Serine/Threonine-rich regions predicted to be hyper-O-glycosylated in the secretory proteins coded by eight fungal genomes

    Directory of Open Access Journals (Sweden)

    González Mario

    2012-09-01

    Full Text Available Abstract Background O-glycosylation of secretory proteins has been found to be an important factor in fungal biology and virulence. It consists in the addition of short glycosidic chains to Ser or Thr residues in the protein backbone via O-glycosidic bonds. Secretory proteins in fungi frequently display Ser/Thr rich regions that could be sites of extensive O-glycosylation. We have analyzed in silico the complete sets of putatively secretory proteins coded by eight fungal genomes (Botrytis cinerea, Magnaporthe grisea, Sclerotinia sclerotiorum, Ustilago maydis, Aspergillus nidulans, Neurospora crassa, Trichoderma reesei, and Saccharomyces cerevisiae in search of Ser/Thr-rich regions as well as regions predicted to be highly O-glycosylated by NetOGlyc (http://www.cbs.dtu.dk. Results By comparison with experimental data, NetOGlyc was found to overestimate the number of O-glycosylation sites in fungi by a factor of 1.5, but to be quite reliable in the prediction of highly O-glycosylated regions. About half of secretory proteins have at least one Ser/Thr-rich region, with a Ser/Thr content of at least 40% over an average length of 40 amino acids. Most secretory proteins in filamentous fungi were predicted to be O-glycosylated, sometimes in dozens or even hundreds of sites. Residues predicted to be O-glycosylated have a tendency to be grouped together forming hyper-O-glycosylated regions of varying length. Conclusions About one fourth of secretory fungal proteins were predicted to have at least one hyper-O-glycosylated region, which consists of 45 amino acids on average and displays at least one O-glycosylated Ser or Thr every four residues. These putative highly O-glycosylated regions can be found anywhere along the proteins but have a slight tendency to be at either one of the two ends.

  4. Motor-mediated bidirectional transport along an antipolar microtubule bundle: A mathematical model

    Science.gov (United States)

    Lin, Congping; Ashwin, Peter; Steinberg, Gero

    2013-05-01

    Long-distance bidirectional transport of organelles depends on the coordinated motion of various motor proteins on the cytoskeleton. Recent quantitative live cell imaging in the elongated hyphal cells of Ustilago maydis has demonstrated that long-range motility of motors and their endosomal cargo occurs on unipolar microtubules (MTs) near the extremities of the cell. These MTs are bundled into antipolar bundles within the central part of the cell. Dynein and kinesin-3 motors coordinate their activity to move early endosomes (EEs) in a bidirectional fashion where dynein drives motility towards MT minus ends and kinesin towards MT plus ends. Although this means that one can easily assign the drivers of bidirectional motion in the unipolar section, the bipolar orientations in the bundle mean that it is possible for either motor to drive motion in either direction. In this paper we use a multilane asymmetric simple exclusion process modeling approach to simulate and investigate phases of bidirectional motility in a minimal model of an antipolar MT bundle. In our model, EE cargos (particles) change direction on each MT with a turning rate Ω and there is switching between MTs in the bundle at the minus ends. At these ends, particles can hop between MTs with rate q1 on passing from a unipolar to a bipolar section (the obstacle-induced switching rate) or q2 on passing in the other direction (the end-induced switching rate). By a combination of numerical simulations and mean-field approximations, we investigate the distribution of particles along the MTs for different values of these parameters and of Θ, the overall density of particles within this closed system. We find that even if Θ is low, the system can exhibit a variety of phases with shocks in the density profiles near plus and minus ends caused by queuing of particles. We discuss how the parameters influence the type of particle that dominates active transport in the bundle.

  5. Influence of commorn smut and anti-tumor screening of substitution lines%玉米染色体片段代换系对瘤黑粉病的抗性研究

    Institute of Scientific and Technical Information of China (English)

    吕爱枝; 王立秋; 曹熙敏; 张晓光

    2011-01-01

    Common smut in maize reduces grain yield greatly. lines play an importment role to minimize the losses caused by common smut. At the elongation stage, Ustilago maydis fungus was injected into 8 maize chromosome segment introgress lines( CSILs). Eight CSILs have different chromosome segments of donor maize HB522 respectively. There are four kinds of varieties in eight CSILs. is highly resist to common smut. Chr3-7's grain yield is highest among other seven CSILs and two parents. chr3-7 is good SSILs for Resistance to common smut study and resistance breeding.%玉米瘤黑粉病能造成玉米严重减产,抗性系的选育对于减少病害具有重要价值.本研究在拔节期采用注射法接种瘤黑粉孢子,对第三染色体的8个染色体片段代换系进行抗性研究.结果表明,8个染色体片段代换系被代换的供体片段位点不同.抗瘤黑粉病表型有4种变异.其中,chr3-7系发病率、病级和病情指数均为0,表现高抗.与其他染色体片段代换系、Z3受体亲本及HB522供体亲本相比,ehr3-7系产量高,接种瘤黑粉孢子与对照产量差异不显著.chr3-7系对于抗瘤黑粉病遗传研究和育种具有重要意义.

  6. The cultural significance of wild mushrooms in San Mateo Huexoyucan, Tlaxcala, Mexico.

    Science.gov (United States)

    Alonso-Aguilar, Luis Enrique; Montoya, Adriana; Kong, Alejandro; Estrada-Torres, Arturo; Garibay-Orijel, Roberto

    2014-03-05

    We performed an ethnomycological study in a community in Tlaxcala, Central Mexico to identify the most important species of wild mushrooms growing in an oak forest, their significance criteria, and to validate the Cultural Significance Index (CSI). Thirty-three mestizo individuals were randomly selected in San Mateo Huexoyucan and were asked seven questions based on criteria established by the CSI. Among the 49 mushroom species collected in the oak forest and open areas, 20 species were mentioned most often and were analyzed in more detail. Ordination and grouping techniques were used to determine the relationship between the cultural significance of the mushroom species, according to a perceived abundance index, frequency of use index, taste score appreciation index, multifunctional food index, knowledge transmission index, and health index. The mushrooms with highest CSI values were Agaricus campestris, Ramaria spp., Amanita aff. basii, Russula spp., Ustilago maydis, and Boletus variipes. These species were characterized by their good taste and were considered very nutritional. The species with the lowest cultural significance included Russula mexicana, Lycoperdon perlatum, and Strobylomyces strobilaceus. The ordination and grouping analyses identified four groups of mushrooms by their significance to the people of Huexoyucan. The most important variables that explained the grouping were the taste score appreciation index, health index, the knowledge transmission index, and the frequency of use index. A. aff. basii and A. campestris were the most significant wild mushrooms to the people of San Mateo. The diversity of the Russula species and the variety of Amanita and Ramaria species used by these people was outstanding. Environments outside the forest also produced useful resources. The CSI used in Oaxaca was useful for determining the cultural significance of mushrooms in SMH, Tlaxcala. This list of mushrooms can be used in conservation proposals for the Quercus

  7. Comparative Analysis Highlights Variable Genome Content of Wheat Rusts and Divergence of the Mating Loci

    Directory of Open Access Journals (Sweden)

    Christina A. Cuomo

    2017-02-01

    Full Text Available Three members of the Puccinia genus, Puccinia triticina (Pt, P. striiformis f.sp. tritici (Pst, and P. graminis f.sp. tritici (Pgt, cause the most common and often most significant foliar diseases of wheat. While similar in biology and life cycle, each species is uniquely adapted and specialized. The genomes of Pt and Pst were sequenced and compared to that of Pgt to identify common and distinguishing gene content, to determine gene variation among wheat rust pathogens, other rust fungi, and basidiomycetes, and to identify genes of significance for infection. Pt had the largest genome of the three, estimated at 135 Mb with expansion due to mobile elements and repeats encompassing 50.9% of contig bases; in comparison, repeats occupy 31.5% for Pst and 36.5% for Pgt. We find all three genomes are highly heterozygous, with Pst [5.97 single nucleotide polymorphisms (SNPs/kb] nearly twice the level detected in Pt (2.57 SNPs/kb and that previously reported for Pgt. Of 1358 predicted effectors in Pt, 784 were found expressed across diverse life cycle stages including the sexual stage. Comparison to related fungi highlighted the expansion of gene families involved in transcriptional regulation and nucleotide binding, protein modification, and carbohydrate degradation enzymes. Two allelic homeodomain pairs, HD1 and HD2, were identified in each dikaryotic Puccinia species along with three pheromone receptor (STE3 mating-type genes, two of which are likely representing allelic specificities. The HD proteins were active in a heterologous Ustilago maydis mating assay and host-induced gene silencing (HIGS of the HD and STE3 alleles reduced wheat host infection.

  8. Evaluation of corn germplasm lines for multiple ear-colonizing insect and disease resistance.

    Science.gov (United States)

    Ni, Xinzhi; Xu, Wenwei; Blanco, Michael H; Wilson, Jeffrey P

    2012-08-01

    Ear-colonizing insects and diseases that reduce yield and impose health threats by mycotoxin contaminations in the grain, are critical impediments for corn (Zea mays L.) production in the southern United States. Ten germplasm lines from the Germplasm Enhancement of Maize (GEM) Program in Ames, IA, and Raleigh, NC, and 10 lines (derived from GEM germplasm) from the Texas Agricultural Experiment Station in Lubbock, TX, were examined in 2007 and 2008 with local resistant and susceptible controls. Four types of insect damage and smut disease (Ustilago maydis) infection, as well as gene X environment (G X E) interaction, was assessed on corn ears under field conditions. Insect damage on corn ears was further separated as cob and kernel damage. Cob penetration rating was used to assess corn earworm [Helicoverpa zea (Boddie)] and fall armyworm [Spodoptera frugiperda (J.E. Smith)] feeding on corn cobs, whereas kernel damage was assessed using three parameters: 1) percentage of kernels discolored by stink bugs (i.e., brown stink bug [Euschistus serous (Say)], southern green stink bug [Nezara viridula (L.)], and green stink bug [Chinavia (Acrosternum) hilare (Say)]; 2) percentage of maize weevil (Sitophilus zeamais Motschulsky)-damaged kernels; and 3) percentage of kernels damaged by sap beetle (Carpophilus spp.), "chocolate milkworm" (Moodna spp.), and pink scavenger caterpillar [Pyroderces (Anatrachyntis) rileyi (Walsingham)]. The smut infection rates on ears, tassels, and nodes also were assessed. Ear protection traits (i.e., husk tightness and extension) in relation to insect damage and smut infection also were examined. Significant differences in insect damage, smut infection, and husk protection traits were detected among the germplasm lines. Three of the 20 germplasm lines were identified as being multiple insect and smut resistant. Of the three lines, entries 5 and 7 were derived from DKXL370, which was developed using corn germplasm from Brazil, whereas entry 14 was

  9. Impact of applying edible oils to silk channels on ear pests of sweet corn.

    Science.gov (United States)

    Ni, Xinzhi; Sparks, Alton N; Riley, David G; Li, Xianchun

    2011-06-01

    The impact of applying edible oils to corn silks on ear-feeding insects in sweet corn, Zea mays L., production was evaluated in 2006 and 2007. Six edible oils used in this experiment were canola, corn, olive, peanut, sesame, and soybean. Water and two commercial insecticidal oils (Neemix neem oil and nC21 Sunspray Ultrafine, a horticultural mineral oil) were used as the controls for the experiment. Six parameters evaluated in this experiment were corn earworm [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] damage rating, the number of sap beetle [Carpophilus spp. (Coleoptera: Nitidulidae)] adults and larvae, the number of corn silk fly (or picture-winged fly) (Diptera: Ulidiidae) larvae, common smut [Ustilago maydis (D.C.) Corda] infection rate, and corn husk coverage. Among the two control treatments, neem oil reduced corn earworm damage at both pre- and postpollination applications in 2006, but not in 2007, whereas the mineral oil applied at postpollination treatments reduced corn earworm damage in both years. The mineral oil also reduced the number of sap beetle adults, whereas the neem oil applied at postpollination attracted the most sap beetle adults in 2007. Among the six edible oil treatments, the corn and sesame oils applied at postpollination reduced corn earworm damage only in 2007. The application of the peanut oil at postpollination attracted more sap beetle adults in 2006, and more sap beetle larvae in 2007. Olive and neem oils significantly reduced husk coverage compared with the water control in both years. The mineral oil application consistently increased smut infection rate in both 2006 and 2007. Ramifications of using oil treatments in ear pest management also are discussed.

  10. Histone deacetylases: revealing the molecular base of dimorphism in pathogenic fungi

    Directory of Open Access Journals (Sweden)

    Alberto Elías-Villalobos

    2015-11-01

    Full Text Available Fungi, as every living organism, interact with the external world and have to adapt to its fluctuations. For pathogenic fungi, such interaction involves adapting to the hostile environment of their host. Survival depends on the capacity of fungi to detect and respond to external stimuli, which is achieved through a tight and efficient genetic control. Chromatin modifications represent a well-known layer of regulation that controls gene expression in response to environmental signals. However, less is known about the chromatin modifications that are involved in fungal virulence and the specific cues and signalling pathways that target chromatin modifications to specific genes. In a recently published study, our research group identified one such regulatory pathway. We demonstrated that the histone deacetylase (HDAC Hos2 is involved in yeast-to-hyphal transition (dimorphism and it is associated with the virulence of the maize pathogen Ustilago maydis, the causative agent of smut disease in corn. Hos2 activates mating-type genes by directly binding to their gene bodies. Furthermore, Hos2 acts downstream of the nutrient-sensing cyclic AMP-Protein Kinase A pathway. We also found that another HDAC, Clr3, contributes to this regulation, possibly in cooperation with Hos2. As a whole, our data suggest that there is a direct link between changes in the environment and acetylation of nucleosomes within certain genes. We propose that histone acetylation is critical to the proper timing and induction of transcription of the genes encoding factors that coordinate changes in morphology with pathogenesis.

  11. The 3-hydroxy-3-methylglutaryl coenzyme-A reductases from fungi: a proposal as a therapeutic target and as a study model.

    Science.gov (United States)

    Andrade-Pavón, Dulce; Sánchez-Sandoval, Eugenia; Rosales-Acosta, Blanca; Ibarra, José Antonio; Tamariz, Joaquín; Hernández-Rodríguez, César; Villa-Tanaca, Lourdes

    2014-01-01

    The enzyme 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR) catalyzes the conversion of HMG-Co-A into mevalonate. This step is the limiting point for the synthesis of cholesterol in mammals and ergosterol in fungi. We describe in this article the genome organization of HMGR coding genes and those deduced from different fungi, recount the evidence showing statins as HMGR inhibitors for ergosterol synthesis and its effect in yeast viability, and propose fungal HMGR (HMGRf) as a model to study the use of pharmaceutical compounds to inhibit cholesterol and ergosterol synthesis. Bibliographical search and bioinformatic analyses were performed and discussed. HMGRfs belong to the class I with a high homology in the catalytic region. The sterol biosynthetic pathway in humans and fungi share many enzymes in the initial steps (such as the HMGR enzyme), but in the last steps enzymes are different rendering the two final products: cholesterol in mammals and ergosterol in fungi. With regards to inhibitors such as statins and other compounds, these affect also fungal viability. Since HMGR from Schizosaccharomyces pombe and Ustilago maydis are very similar to the human HMGR in the catalytic regions, we propose that fungal enzymes can be used to test inhibitors for a potential use in humans. We consider that HMGRf is a good therapeutic target to design and test new antifungal compounds. 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). Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  12. Comparative EST analysis provides insights into the basal aquatic fungus Blastocladiella emersonii

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    Gomes Suely L

    2006-07-01

    Full Text Available Abstract Background Blastocladiella emersonii is an aquatic fungus of the Chytridiomycete class, which is at the base of the fungal phylogenetic tree. In this sense, some ancestral characteristics of fungi and animals or fungi and plants could have been retained in this aquatic fungus and lost in members of late-diverging fungal species. To identify in B. emersonii sequences associated with these ancestral characteristics two approaches were followed: (1 a large-scale comparative analysis between putative unigene sequences (uniseqs from B. emersonii and three databases constructed ad hoc with fungal proteins, animal proteins and plant unigenes deposited in Genbank, and (2 a pairwise comparison between B. emersonii full-length cDNA sequences and their putative orthologues in the ascomycete Neurospora crassa and the basidiomycete Ustilago maydis. Results Comparative analyses of B. emersonii uniseqs with fungi, animal and plant databases through the two approaches mentioned above produced 166 B. emersonii sequences, which were identified as putatively absent from other fungi or not previously described. Through these approaches we found: (1 possible orthologues of genes previously identified as specific to animals and/or plants, and (2 genes conserved in fungi, but with a large difference in divergence rate in B. emersonii. Among these sequences, we observed cDNAs encoding enzymes from coenzyme B12-dependent propionyl-CoA pathway, a metabolic route not previously described in fungi, and validated their expression in Northern blots. Conclusion Using two different approaches involving comparative sequence analyses, we could identify sequences from the early-diverging fungus B. emersonii previously considered specific to animals or plants, and highly divergent sequences from the same fungus relative to other fungi.

  13. Dicty_cDB: Contig-U11279-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ( FG251718 ) CBYF1589.g1 CBYF Cercospora zeae-maydis conidiati... 52 0.067 1 ( FG250521 ) CBYC640.g1 CBYC Cercospora... zeae-maydis cercospori... 52 0.067 1 ( FG243214 ) CBYB2040.g1 CBYB Cercospora zeae-maydis cercospor

  14. Microsporogenesis and pollen grains in Silene dioica (L. Cl. and alterations in its anthers parasited by Ustilago violacea (Pers. Rouss. (Ustilaginales

    Directory of Open Access Journals (Sweden)

    Jean-Claude Audran

    2014-01-01

    Full Text Available Healthy and infected anthers are comparatively studied with optical and electron microscopic techniques. The fungus stops the stamen histogenesis at an early stage and destroys specifically the sporogenous tissue.

  15. Microsporogenesis and pollen grains in Silene dioica (L.) Cl. and alterations in its anthers parasited by Ustilago violacea (Pers.) Rouss. (Ustilaginales)

    OpenAIRE

    Jean-Claude Audran; Mekinto Batcho

    2014-01-01

    Healthy and infected anthers are comparatively studied with optical and electron microscopic techniques. The fungus stops the stamen histogenesis at an early stage and destroys specifically the sporogenous tissue.

  16. Evaluation of resistance in grasses to Gaeumannomyces graminis var. maydis%禾草对禾顶囊壳玉米变种的抗病性研究

    Institute of Scientific and Technical Information of China (English)

    石仁才; 商鸿生

    2006-01-01

    利用人工接菌方法测定了10种草坪禾草,包括多年生黑麦草、高羊茅、紫羊茅、中华结缕草、硬羊茅、草地早熟禾、加拿大早熟禾、匍匐剪股颖、细弱剪股颖、狗牙根;13种禾本科牧草,包括一年生黑麦草、鸭茅、碱茅、扁穗冰草、沙生冰草、披碱草、老芒麦、杂交小米草、猫尾草、弯叶画眉草、无芒雀麦、百喜草、苏丹草;以及4种禾本科作物,包括燕麦、甜高粱、小麦、玉米对禾顶囊壳玉米变种的抗病性.结果表明,多数供试禾草对其具有免疫能力,多年生黑麦草、一年生黑麦草、草地早熟禾、小麦高度抗侵染;无芒雀麦和玉米具有中等程度的抗侵染能力,但抗侵入能力较差;高粱属的苏丹草和甜高粱中度感病,属于中感作物.多年生黑麦草和草地早熟禾品种间存在着抗病性差异,有免疫品种和高抗品种.

  17. 玉米须多糖的利尿作用研究%Study on diuretic effect of stigma maydis polysaccharide

    Institute of Scientific and Technical Information of China (English)

    窦传斌; 杜娟; 许启泰

    2007-01-01

    目的:研究玉米须多糖(SMPS)的利尿作用.方法:采用代谢笼法和滤纸法测定NS负荷大、小鼠的排尿量及尿中Na+、K+、Cl-的含量.采用甘油致大鼠肾衰竭模型观察SMPS对肾衰竭大鼠排尿的影响.结果:高、中剂量的SMPS可明显增加大、小鼠的排尿量,增加尿液中K+、Cl-的含量,对Na+含量无明显影响;增肾衰竭大鼠的排尿量.结论:玉米须多糖具有明显的利尿作用.

  18. 台湾Pseudozyma属之分类学研究

    Institute of Scientific and Technical Information of China (English)

    刘桂郁; 魏育慧; 李福临; 许文浩; 陈学如; 陈建州; 温秋燕; 林锡杰; 朱文深; 袁国芳

    2004-01-01

    黑穗菌目(Ustilaginales)感染重要作物造成重大之经济损失,其中黑穗菌属(Ustilago)大部分的种可寄生于单子叶植物,是黑穗菌目最大的一个属;黑穗菌目之无性世代曾被归群至不同的属,例如:Candida,Pseudozyma,Sporobolomyces,Sterigmatomyces,Stephanoascus,及Trichosporon等,至1995年,Boekhout等学者依据形态、生理生化特性以及26S rDNA D1&D2片段分析,将这些种重新分类,归属于Pseudozyma,并以26S rDNA D1&D2片段之分析结果,确认Pseudozyma属与玉米黑穗病(Ustilago maydis)之类缘关系.Pseudozyma属为类酵母真菌,至1998年为止,所有的种均分离自植物组织,例如叶、花及果实,共有七个种;但是在2003年,Sugita等学者报导在泰国自病人的血液样品中也分离到Pseudozyma属,其中两个菌株为新种,目前,Pseudozyma属共有九个种,由文献之记载可知,其分布于南极、泰国、日本、德国、加拿大、英国及巴西等区域.在台湾,自合欢(Albizia julibrissin)、杏梅(Prunus mume)及玫瑰(Rosa rugosa)等植物的花分离到Pseudozyma属之菌株,已寄存于生物资源保存及研究中心,编号分别为BCRC 33871、33950、33951,依据形态特征、生理生化特性及rDNA ITS1-5.8S-ITS2片段序列分析进行鉴定,BCRC 33871种名为P.antarctica,是台湾新纪录种;BCRC 33950及33951则可能为新种,尚待进一步之发表.此外,本研究利用生理生化特性及rDNA序列分析探讨P.antarctica种内与其近似种P.rugulosa及P.aphidis间的差异,并讨论用以区分P.antarctica与近似种之关键性特征.

  19. Genome-Wide Annotation and Comparative Analysis of Cytochrome P450 Monooxygenases in Basidiomycete Biotrophic Plant Pathogens.

    Directory of Open Access Journals (Sweden)

    Lehlohonolo Benedict Qhanya

    Full Text Available Fungi are an exceptional source of diverse and novel cytochrome P450 monooxygenases (P450s, heme-thiolate proteins, with catalytic versatility. Agaricomycotina saprophytes have yielded most of the available information on basidiomycete P450s. This resulted in observing similar P450 family types in basidiomycetes with few differences in P450 families among Agaricomycotina saprophytes. The present study demonstrated the presence of unique P450 family patterns in basidiomycete biotrophic plant pathogens that could possibly have originated from the adaptation of these species to different ecological niches (host influence. Systematic analysis of P450s in basidiomycete biotrophic plant pathogens belonging to three different orders, Agaricomycotina (Armillaria mellea, Pucciniomycotina (Melampsora laricis-populina, M. lini, Mixia osmundae and Puccinia graminis and Ustilaginomycotina (Ustilago maydis, Sporisorium reilianum and Tilletiaria anomala, revealed the presence of numerous putative P450s ranging from 267 (A. mellea to 14 (M. osmundae. Analysis of P450 families revealed the presence of 41 new P450 families and 27 new P450 subfamilies in these biotrophic plant pathogens. Order-level comparison of P450 families between biotrophic plant pathogens revealed the presence of unique P450 family patterns in these organisms, possibly reflecting the characteristics of their order. Further comparison of P450 families with basidiomycete non-pathogens confirmed that biotrophic plant pathogens harbour the unique P450 families in their genomes. The CYP63, CYP5037, CYP5136, CYP5137 and CYP5341 P450 families were expanded in A. mellea when compared to other Agaricomycotina saprophytes and the CYP5221 and CYP5233 P450 families in P. graminis and M. laricis-populina. The present study revealed that expansion of these P450 families is due to paralogous evolution of member P450s. The presence of unique P450 families in these organisms serves as evidence of how a host

  20. Genome-Wide Annotation and Comparative Analysis of Cytochrome P450 Monooxygenases in Basidiomycete Biotrophic Plant Pathogens.

    Science.gov (United States)

    Qhanya, Lehlohonolo Benedict; Matowane, Godfrey; Chen, Wanping; Sun, Yuxin; Letsimo, Elizabeth Mpholoseng; Parvez, Mohammad; Yu, Jae-Hyuk; Mashele, Samson Sitheni; Syed, Khajamohiddin

    2015-01-01

    Fungi are an exceptional source of diverse and novel cytochrome P450 monooxygenases (P450s), heme-thiolate proteins, with catalytic versatility. Agaricomycotina saprophytes have yielded most of the available information on basidiomycete P450s. This resulted in observing similar P450 family types in basidiomycetes with few differences in P450 families among Agaricomycotina saprophytes. The present study demonstrated the presence of unique P450 family patterns in basidiomycete biotrophic plant pathogens that could possibly have originated from the adaptation of these species to different ecological niches (host influence). Systematic analysis of P450s in basidiomycete biotrophic plant pathogens belonging to three different orders, Agaricomycotina (Armillaria mellea), Pucciniomycotina (Melampsora laricis-populina, M. lini, Mixia osmundae and Puccinia graminis) and Ustilaginomycotina (Ustilago maydis, Sporisorium reilianum and Tilletiaria anomala), revealed the presence of numerous putative P450s ranging from 267 (A. mellea) to 14 (M. osmundae). Analysis of P450 families revealed the presence of 41 new P450 families and 27 new P450 subfamilies in these biotrophic plant pathogens. Order-level comparison of P450 families between biotrophic plant pathogens revealed the presence of unique P450 family patterns in these organisms, possibly reflecting the characteristics of their order. Further comparison of P450 families with basidiomycete non-pathogens confirmed that biotrophic plant pathogens harbour the unique P450 families in their genomes. The CYP63, CYP5037, CYP5136, CYP5137 and CYP5341 P450 families were expanded in A. mellea when compared to other Agaricomycotina saprophytes and the CYP5221 and CYP5233 P450 families in P. graminis and M. laricis-populina. The present study revealed that expansion of these P450 families is due to paralogous evolution of member P450s. The presence of unique P450 families in these organisms serves as evidence of how a host

  1. In silico prediction and characterization of secondary metabolite biosynthetic gene clusters in the wheat pathogen Zymoseptoria tritici.

    Science.gov (United States)

    Cairns, Timothy; Meyer, Vera

    2017-08-17

    Fungal pathogens of plants produce diverse repertoires of secondary metabolites, which have functions ranging from iron acquisition, defense against immune perturbation, to toxic assaults on the host. The wheat pathogen Zymoseptoria tritici causes Septoria tritici blotch, a foliar disease which is a significant threat to global food security. Currently, there is limited knowledge of the secondary metabolite arsenal produced by Z. tritici, which significantly restricts mechanistic understanding of infection. In this study, we analyzed the genome of Z. tritici isolate IP0323 to identify putative secondary metabolite biosynthetic gene clusters, and used comparative genomics to predict their encoded products. We identified 32 putative secondary metabolite clusters. These were physically enriched at subtelomeric regions, which may facilitate diversification of cognate products by rapid gene rearrangement or mutations. Comparative genomics revealed a four gene cluster with significant similarity to the ferrichrome-A biosynthetic locus of the maize pathogen Ustilago maydis, suggesting this siderophore is deployed by Z. tritici to acquire iron. The Z. tritici genome also contains several isoprenoid biosynthetic gene clusters, including one with high similarity to a carotenoid/opsin producing locus in several fungi. Furthermore, we identify putative phytotoxin biosynthetic clusters, suggesting Z. tritici can produce an epipolythiodioxopiperazine, and a polyketide and non-ribosomal peptide with predicted structural similarities to fumonisin and the Alternaria alternata AM-toxin, respectively. Interrogation of an existing transcriptional dataset suggests stage specific deployment of numerous predicted loci during infection, indicating an important role of these secondary metabolites in Z. tritici disease. We were able to assign putative biosynthetic products to numerous clusters based on conservation amongst other fungi. However, analysis of the majority of secondary

  2. Knowledge and use of edible mushrooms in two municipalities of the Sierra Tarahumara, Chihuahua, Mexico.

    Science.gov (United States)

    Quiñónez-Martínez, Miroslava; Ruan-Soto, Felipe; Aguilar-Moreno, Ivonne Estela; Garza-Ocañas, Fortunato; Lebgue-Keleng, Toutcha; Lavín-Murcio, Pablo Antonio; Enríquez-Anchondo, Irma Delia

    2014-09-17

    The Sierra Madre Occidental of Chihuahua in Northern Mexico is inhabited by indigenous Raramuris, mestizos, and other ethnic groups. The territory consists of canyons and ravines with pine, oak and pine-oak forests in the higher plateaus. A great diversity of potentially edible mushrooms is found in forests of the Municipalities of Bocoyna and Urique. Their residents are the only consumers of wild mushrooms in the Northern Mexico; they have a long tradition of collecting and eating these during the "rainy season." However, despite the wide diversity of edible mushrooms that grow in these areas, residents have a selective preference. This paper aims to record evidence of the knowledge and use of wild potentially edible mushroom species by inhabitants of towns in the Sierra Tarahumara of Chihuahua, Mexico. Using a semi-structured technique, we surveyed 197 habitants from seven locations in Urique, Bocoyna, and the Cusarare area from 2010 to 2012. Known fungi, local nomenclature, species consumed, preparation methods, appreciation of taste, forms of preservation, criteria for differentiating toxic and edible fungi, other uses, economic aspects, and traditional teaching were recorded. To identify the recognized species, photographic stimuli of 22 local edible species and two toxic species were used. The respondents reported preference for five species: Amanita rubescens, Agaricus campestris, Ustilago maydis, Hypomyces lactifluorum, and the Amanita caesarea complex. No apparent differences were found between ethnic groups in terms of preference, although mestizos used other species in Bocoyna (Boletus edulis and B. pinophilus). Some different uses of fungi are recognized by respondents, i.e. home decorations, medicine, as food in breeding rams, etc. The studied population shows a great appreciation towards five species, mainly the A. caesarea complex, and an apparent lack of knowledge of nearly 20 species which are used as food in other areas of Mexico. There are no

  3. A kingdom-specific protein domain HMM library for improved annotation of fungal genomes

    Directory of Open Access Journals (Sweden)

    Oliver Stephen G

    2007-04-01

    Full Text Available Abstract Background Pfam is a general-purpose database of protein domain alignments and profile Hidden Markov Models (HMMs, which is very popular for the annotation of sequence data produced by genome sequencing projects. Pfam provides models that are often very general in terms of the taxa that they cover and it has previously been suggested that such general models may lack some of the specificity or selectivity that would be provided by kingdom-specific models. Results Here we present a general approach to create domain libraries of HMMs for sub-taxa of a kingdom. Taking fungal species as an example, we construct a domain library of HMMs (called Fungal Pfam or FPfam using sequences from 30 genomes, consisting of 24 species from the ascomycetes group and two basidiomycetes, Ustilago maydis, a fungal pathogen of maize, and the white rot fungus Phanerochaete chrysosporium. In addition, we include the Microsporidion Encephalitozoon cuniculi, an obligate intracellular parasite, and two non-fungal species, the oomycetes Phytophthora sojae and Phytophthora ramorum, both plant pathogens. We evaluate the performance in terms of coverage against the original 30 genomes used in training FPfam and against five more recently sequenced fungal genomes that can be considered as an independent test set. We show that kingdom-specific models such as FPfam can find instances of both novel and well characterized domains, increases overall coverage and detects more domains per sequence with typically higher bitscores than Pfam for the same domain families. An evaluation of the effect of changing E-values on the coverage shows that the performance of FPfam is consistent over the range of E-values applied. Conclusion Kingdom-specific models are shown to provide improved coverage. However, as the models become more specific, some sequences found by Pfam may be missed by the models in FPfam and some of the families represented in the test set are not present in FPfam

  4. 脱乙酰壳多糖抑制真菌生长的构效关系%Structure-Function Relationship of Chitosan against Phytopathogenic Fungi

    Institute of Scientific and Technical Information of China (English)

    杨典洱; 林晓怡; 向本琼; 杨照罡; 王海龙; 魏群

    2005-01-01

    本研究目标是研究脱乙酰壳多糖的化学结构(乙酰化程度DA和聚合程度DP)与它的抑制真菌生长能力之间的构效关系.选用了12个分属于3个系列、化学结构相关而又不同的、结构清晰的脱乙酰壳多糖和3种不同的真菌(Fusarium solani,Fusarium graminearum和Ustilago maydis).通过分别测定每个脱乙酰壳多糖对3种真菌的生长曲线和最低抑制浓度(MIC,minimum inhibitory concentration);比较各个系列脱乙酰壳多糖的MIC和它的化学结构(DA和DP)之间的关系.结果显示对同一种真菌,不同脱乙酰壳多糖的抑制真菌生长曲线形态和MIC是各不相同的;同样同一脱乙酰壳多糖,对不同真菌也有其特殊的生长曲线和MIC;通常随着脱乙酰壳多糖中DA的递增,MIC是增加的,其抑制真菌的活性是降低的;在DA相同的条件下,随着DP的递增,MIC也是增加的,其抑制真菌的活性是减低的.所以可以说,脱乙酰壳多糖抑制真菌生长的能力与其化学结构紧密相关,在本实验的条件下,脱乙酰壳多糖分子越小,分子中的自由氨基越多,抑制真菌的活性越大.

  5. Gene discovery in EST sequences from the wheat leaf rust fungus Puccinia triticina sexual spores, asexual spores and haustoria, compared to other rust and corn smut fungi

    Directory of Open Access Journals (Sweden)

    Wynhoven Brian

    2011-03-01

    . tritici (Pst, and poplar leaf rust Melampsora species, and the corn smut fungus, Ustilago maydis (Um. While extensive homologies were found, many genes appeared novel and species-specific; over 40% of genes did not match any known sequence in existing databases. Focusing on spore stages, direct comparison to Um identified potential functional homologs, possibly allowing heterologous functional analysis in that model fungus. Many potentially secreted protein genes were identified by similarity searches against genes and proteins of Pgt and Melampsora spp., revealing apparent orthologs. Conclusions The current set of Pt unigenes contributes to gene discovery in this major cereal pathogen and will be invaluable for gene model verification in the genome sequence.

  6. Dicty_cDB: Contig-U05491-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ADI_5th_i... 46 1.9 1 ( FG250469 ) CBYC608.b1 CBYC Cercospora zeae-maydis cercosp...ori... 46 1.9 1 ( FG246443 ) CBYB4322.b1 CBYB Cercospora zeae-maydis cercospor... 46 1.9 1 ( FG244122 ) CBYB2657.g1 CBYB Cercospora

  7. Dicty_cDB: Contig-U10097-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available tyostelium discoideum slug cDNA, clone SSL239. 307 e-108 2 ( FG252229 ) CBYF541.g1 CBYF Cercospora... zeae-maydis conidiatio... 58 3e-18 4 ( FG252228 ) CBYF541.b1 CBYF Cercospora zeae-maydis c

  8. Datos preliminares de la evaluación del mecanismo de tolerancia en trigo a Sipha maydis mediante el uso del ácido jasmónico

    OpenAIRE

    Vicente, Augusto P.; Sartori, Aldo; Marro, Eduardo; Saldúa, Vilma Luciana

    2015-01-01

    Las plantas han desarrollado distintos sistemas de defensas ante el estrés biótico. Las defensas constitutivas, son preexistentes al ataque del patógeno o de los insectos. Las defensas inducidas son activadas de manera local ante el ataque del patógeno y/o insecto, están mediadas por fitohormonas, entre ellas el ácido Jasmónico (AJ), hormona que modula la acción de genes implicados en las defensas y en varios procesos del desarrollo de la planta. La tolerancia es un tipo de resistencia que ot...

  9. 立枯丝核菌(水稻纹枯病菌)G蛋白β亚基基因的克隆与特性分析%Cloning and expression of G-protein beta-subunit in rice Rhizoctonia solani

    Institute of Scientific and Technical Information of China (English)

    曲广林; 李仕贵; 徐正君; 王玉平; 黄文娟; 林瑜凡; 万佳; 马炳田

    2008-01-01

    由立枯丝核菌Rhizoctonia solani引起的水稻纹枯病(rice sheath blight)是水稻三大病害之一.G蛋白β亚基(G-protein beta-subunit)编码的蛋白作为重要的信号传导蛋白,在其致病分子机制中起着重要作用.为了解G蛋白β亚基基因的作用方式,本文根据同源物种G蛋白β亚基相关序列设计引物,通过PCR(polymerase chain reaction)和RT-PCR(reverse transcriptase PCR)技术,获得了以水稻为寄主的立枯丝核菌G蛋白β亚基(G-protein beta-subunit of rice Rhizoctonia solani,简写gbrrs1)的基因序列和开放阅读框ORF(open reading frame)(GenBank登录号EU267677).该基因全长1867bp,含有4个内含子和5个外显子,各内含子长度在54bp-65bp,且序列均符合5'-gt……ag-3,模式.开放阅读框1047bp,编码348aa,推测的蛋白质分子量为38.23kDa,等电点为6.54.该蛋白质具有2个alpha-helix和7个beta sheet的二级结构,每个beta sheet又包含4个beta-strand.gbrrs1在N端有2个alpha-helix,紧接着是由7个beta sheet由无规则卷曲连接形成的桶形结构.GenBank Blast结果表明,gbrrsl与四种真菌生物G蛋白β亚基的氨基酸序列同源性较高,与Lentinula edodes(AAT74567.1)、Coprinopsis cinema(EAU92269)、Ustilago maydis(AAN33051)和Filobasidiella neoformans(AAD03596)的一致性分别达到了89%、88%、81%和81%.将gbrss1的开放阅读框克隆于原核融合表达载体pGEX-4T-2中,经异丙基-β-D-硫代半乳糖苷(IPTG)诱导,获得了相应蛋白的表达.

  10. Dicty_cDB: Contig-U06911-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 63 ) 7LEAF--06-J20.b1 Rice leaf plasmid cDNA library I... 46 1.2 1 ( FG253080 ) CBYG1302.g1 CBYG Cercospora ...zeae-maydis conidiati... 46 1.2 1 ( FG253079 ) CBYG1302.b1 CBYG Cercospora zeae-m...aydis conidiati... 46 1.2 1 ( FG243788 ) CBYB2420.g1 CBYB Cercospora zeae-maydis cercospor... 46 1.2 1 ( FD7

  11. Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens

    KAUST Repository

    Ali, Shawkat

    2014-09-19

    The genetic transformation of certain organisms, required for gene function analysis or complementation, is often not very efficient, especially when dealing with large gene constructs or genomic fragments. We have adapted the natural DNA transfer mechanism from the soil pathogenic bacterium Agrobacterium tumefaciens, to deliver intact large DNA constructs to basidiomycete fungi of the genus Ustilago where they stably integrated into their genome. To this end, Bacterial Artificial Chromosome (BAC) clones containing large fungal genomic DNA fragments were converted via a Lambda phage-based recombineering step to Agrobacterium transfer-competent binary vectors (BIBACs) with a Ustilago-specific selection marker. The fungal genomic DNA fragment was subsequently successfully delivered as T-DNA through Agrobacterium-mediated transformation into Ustilago species where an intact copy stably integrated into the genome. By modifying the recombineering vector, this method can theoretically be adapted for many different fungi.

  12. Dicty_cDB: Contig-U01723-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available cospora zeae-maydis cercospor... 32 5.4 2 ( GD236757 ) G1045P366RL10.T1 Aplysia cal...EY383727 ) CAXA476.fwd CAXA Helobdella robusta Subtracted La... 38 5.4 2 ( FG247080 ) CBYB4735.b1 CBYB Cer

  13. AcEST: DK950768 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ospora zeae-maydis GN=CZK3 PE=4 SV=1 Length = 1372 Score = 35.4 bits (80), Expect =...HLH 206 Query: 54 GA 49 GA Sbjct: 207 GA 208 >tr|Q7Z9J3|Q7Z9J3_9PEZI MAP kinase kinase kinase Czk3 OS=Cerc

  14. Dicty_cDB: Contig-U15661-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 8 ) SCCCLR1024F06.g LR1 Saccharum officinarum cDNA cl... 76 5e-09 1 ( FG245328 ) CBYB3528.b1 CBYB Cercospora... zeae-maydis cercospor... 76 5e-09 1 ( FG242556 ) CBYB1587.b1 CBYB Cercospora zea

  15. Dicty_cDB: Contig-U12034-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2365 ) Mus musculus mammary gland RCB-0526 Jyg-MC(A) cDN... 48 0.80 1 ( FG244122 ) CBYB2657.g1 CBYB Cercos...pora zeae-maydis cercospor... 48 0.80 1 ( EK182641 ) 1095458129632 Global-Ocean-Sam

  16. Survival of Stenocarpella spp. in maize debris and soil suppressiveness to maize ear rot pathogens

    NARCIS (Netherlands)

    Moretti Ferreira Pinto, Felipe; Novaes Medeiros, H.; Biazzotto Correia Porto, V.; Silva Siqueira, da C.; Cruz Machado, da J.; Köhl, J.; Vasconcelos de Medeiros, Flavio

    2016-01-01

    Stenocarpella species (S. maydis and S. macrospora) overwinter saprophytically in maize stubble but little is known about the factors that contribute to its survival and to the induction of suppressiveness of pathogen colonization. We aimed at determining the role of crop rotation on the survival of

  17. Identification of a chitinase modifying protein from Fusarium verticillioides: truncation of a host resistance protein by a fungalysin metalloprotease

    Science.gov (United States)

    Chitinase modifying proteins (cmps) are proteases, secreted by fungal pathogens, which truncate the plant class IV chitinases ChitA and ChitB during maize ear rot. Cmp activity has been characterized for Bipolaris zeicola and Stenocarpella maydis, but the identities of the proteases are not known. H...

  18. 7 CFR 319.24 - Notice of quarantine.

    Science.gov (United States)

    2010-01-01

    ... SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Corn Diseases Quarantine § 319.24 Notice of..., that maize or Indian corn (Zea mays L.) and closely related plants are subject to certain injurious diseases, especially Peronospora maydis Raciborski, Sclerospora sacchari Miyake and other downy...

  19. Incidence and Severity of Maize Ear Rots and Factors Responsible for Their Occurrence in Uganda

    Science.gov (United States)

    Bigirwa, G.; Kaaya, A. N.; Sseruwu, G.; Adipala, E.; Okanya, S.

    Eleven major maize growing districts of Uganda were surveyed for three consecutive seasons between 2002 and 2003 to establish maize ear rot incidence and severity. Sternocarpella maydis and Fusarium species particularly F. graminearum and F. verticillioides were the identified maize ear rot causing fungi. Incidence of S. maydis ranged from 2.5 to 32.5% while that of Fusarium sp. was in the range of 1.9 and 15.3%. In districts of higher altitude (above 1,800 m above sea level) F. graminearum dominated in all seasons while in districts with an altitude between 900 and 1,500 m above sea level, S. maydis was the major cause of ear rots. This observation was attributed to differences in temperature, altitude and rainfall. There was a strong positive correlation (p = 0.001) between incidence and severity for S. maydis and a weak one for Fusarium sp. because the latter would rarely infect the entire cob unlike the former. All farmers expressed concern about the quality of maize due to ear rots and sort out infected grain after harvest. However, varied uses of infected grain were noted. For example, in Kapchorwa district 82% of the respondents indicated that the infected grain is used for making local brew because the moulds give it a good taste and aroma, while in Kamuli and Masaka districts, 36% use it as animal feed ingredient. This indicates that people and animals could be ingesting mycotoxins unknowingly thus the need for sensitization programmes.

  20. Significant yield increases from control of leaf diseases in maize - an overlooked problem?!

    DEFF Research Database (Denmark)

    Jørgensen, Lise Nistrup

    2012-01-01

    The area of maize has increased in several European countries in recent years. In Denmark, the area has increased from 10,000 ha in 1980 to 185,000 ha in 2011. Initially only silage maize was cultivated in Denmark, but in more recent years the area of grain maize has also increased. Farms growing...... maize have often grown maize consecutively for several years. This leads to significant amounts of debris building up in the fields, which serves as an important source of inoculum for diseases when new crops are grown. In recent years, leaf diseases have caused production problems over several seasons....... Two major diseases have been identified: Eyespot (Kabatiella zeae) and Northern leaf blotch (Exserohilum turcicum). Other diseases, including Southern maize leaf blight or maydis leaf blight, caused by Bipolaris maydis, and Northern corn leaf spot, caused by Bipolaris zeicola, may potentially play...

  1. Comparison of fungi within the Gaeumannomyces-Phialophora complex by analysis of ribosomal DNA sequences.

    OpenAIRE

    Bryan, G. T.; Daniels, M. J.; Osbourn, A E

    1995-01-01

    Four ascomycete species of the genus Gaeumannomyces infect roots of monocotyledons. Gaeumannomyces graminis contains four varieties, var. tritici, var. avenae, var. graminis, and var. maydis. G. graminis varieties tritici, avenae, and graminis have Phialophora-like anamorphs and, together with the other Gaeumannomyces and Phialophora species found on cereal roots, constitute the Gaeumannomyces-Phialophora complex. Relatedness of a number of Gaeumannomyces and Phialophora isolates was assessed...

  2. N,N{prime}-dicyclohexylcarbodiimide cross-linking suggests a central core of helices II in oligomers of URF13, the pore-forming T-toxin receptor of cms-T maize mitochondria

    Energy Technology Data Exchange (ETDEWEB)

    Rhoads, D.M. [Duke Univ., Durham, NC (United States)]|[North Carolina State Univ., Raleigh, NC (United States); Kaspi, C.I.; Siedow, J.N. [Duke Univ., Durham, NC (United States); Levings, C.S. III [North Carolina State Univ., Raleigh, NC (United States)

    1994-08-16

    URF13 is a mitochondrially encoded, integral membrane protein found only in maize carrying the cms-T cytoplasm. URF13 is associated with cytoplasmic male sterility, Texas type, and causes susceptibility to the fungal pathogens Bipolaris maydis race T and Phyllosticta maydis. URF13 is predicted to contain three transmembrane {alpha}-helices and is a receptor for the pathotoxins (T-toxins) produced by B. maydis race T and P. maydis. Binding of T-toxin to URF13 leads to membrane permeability. Cross-linking of URF13 oligomers with N,N{prime}-dicyclohexylcarbodiimide (DCCD) protects Escherichia coli cells expressing URF13 and cms-T mitochondria from the permeability caused by T-toxin or methomyl. Using mutated forms of URF13 expressed in E. coli cells, the authors determined the molecular mechanism of DCCD protection. They separately changed Lys-37 in helix II to isoleucine (K37I-URF13) and Lys-32 in the helix I/helix II loop region to alanine (K32A-URF13). DCCD treatment of K37I-URF13-expressing cells did not protect the cells from permeability caused by T-toxin or methomyl. DCCD cross-linking was greatly reduced in K37I-URF13 and in D39V-URF13-expressing cells, but it was unaffected in K32A-URF13-expressing cells. Binding of methomyl or T-toxin decreases DCCD cross-linking of URF13 oligomers expressed in either E. coli or cms-T mitochondria. They conclude that Asp-39 in helix II is cross-linked by DCCD to Lys-37 in helix II of an adjacent URF13 molecule and that this cross-linking protects against toxin-mediated permeabilization. The results also indicate that helices II form a central core in URF13 oligomers.

  3. 沿淮地区玉米病害初步鉴定

    Institute of Scientific and Technical Information of China (English)

    王波; 史玲莉

    2003-01-01

    对沿淮地区玉米病害调查采样,分离培养,鉴定.结果表明该地区发生的玉米病害有十几种.主要病害是:玉米纹枯病(Rhizoctonia solani),玉米瘤黑粉病(Vstilago maydis)及各种叶斑病.还首次发现玉米细菌性茎腐病(Erwinia carotovora).

  4. Essential oils on the control of stem and ear rot in maize

    Directory of Open Access Journals (Sweden)

    Glauco Antonio Teixeira

    2013-11-01

    Full Text Available Stem and ear rot caused by Stenocarpella maydis are responsible for severe losses in maize production. Treatment of seeds with fungicides may induce environmental damage. Hence, this study aimed to evaluate the effects of essential oils extracted from Cymbopogon winterianus, Thymus vulgaris, Cymbopogon citratus, Corymbia citriodora, Cinnamomum zeylanicum, and Syzygium aromaticum on the development of in vitro S. maydis. In addition, maize seeds were treated with these essential oils to determine their possible mode of action and effects. The oils from S. aromaticum, C. zeylanicum, and T. vulgaris inhibited fungal development at concentrations higher than 0.025%. The oils from S. aromaticum and C. zeylanicum showed seed germination rates of 89.0% and 84.5%, which were higher than that of the control. The oils from S. aromaticum and C. zeylanicum reduced the pathogen incidence in the seeds to 39.0% and 28.0%, respectively. Further, these oils as well as that from T. vulgaris produced lower reduction of maize stand. Scanning electron microscopy examination revealed that essential oils from S. aromaticum and T. vulgaris acted directly on the conidia, impeding germination. The findings suggest that the oils from S. aromaticum, C. zeylanicum, and T. vulgaris are potential alternatives for maize seed treatment in the control of S. maydis.

  5. Indirect selection for resistance to ear rot and leaf diseases in maize lines using biplots.

    Science.gov (United States)

    Pereira, G S; Camargos, R B; Balestre, M; Von Pinho, R G; C Melo, W M

    2015-09-21

    Leaf disease and ear rot have caused reductions in maize yield in Brazil and other producer countries. Therefore, the aims of this study were to analyze the association between husked ear yield and the severity of maize white spot, gray leaf spot, helminthosporium, and ear rot caused by Fusarium verticillioides and Diplodia maydis using biplots in a mixed-model approach. The responses of 238 lines introduced to Brazil and four controls were evaluated using an incomplete block design with three replicates in two locations: Lavras and Uberlândia, Minas Gerais, Brazil. Two experiments were conducted in each location, one with F. verticillioides and the other with D. maydis. The mixed models elucidated the relationship between yield, leaf disease, and ear disease. Significant genotype x environment and genotype x pathogen interactions were observed. In conclusion, husked ear yield is more associated with ear rot than with the leaf diseases evaluated, justifying the indirect selection for resistance to kernel rot in maize-F. verticillioides and maize-D. maydis pathosystems by yield evaluation.

  6. Detecção de espécies de Fusarium potencialmente produtoras de Micotoxinas em grãos de milho no Nordeste do Brasil.

    OpenAIRE

    Maruzanete Pereira Melo

    2011-01-01

    O Brasil é o terceiro maior produtor mundial de grãos milho (Zea mays L.), ficando atrás apenas dos Estados Unidos e China. Os agentes causadores de podridões de espiga e colmo são fungos como: Stenocarpella maydis, Fusarium spp e Penicillium oxalicum, merecendo destaque para as espécies de Fusarium, em especial as espécies do complexo Giberrela fujikuroi. As espécies do complexo, frequetemente associado à podridão de colmo e espiga são: Fusarium verticillioides (Gibberella moniliformis), Fus...

  7. Detecção de espécies de Fusarium potencialmente produtoras de Micotoxinas em grãos de milho no Nordeste do Brasil.

    OpenAIRE

    Maruzanete Pereira de Melo

    2011-01-01

    O Brasil é o terceiro maior produtor mundial de grãos milho (Zea mays L.), ficando atrás apenas dos Estados Unidos e China. Os agentes causadores de podridões de espiga e colmo são fungos como: Stenocarpella maydis, Fusarium spp e Penicillium oxalicum, merecendo destaque para as espécies de Fusarium, em especial as espécies do complexo Giberrela fujikuroi. As espécies do complexo, frequetemente associado à podridão de colmo e espiga são: Fusarium verticillioides (Gibberella moniliformis), Fus...

  8. Detection of fungal infectous agent of wheat grains in store-pits of Markazi province, Iran.

    Science.gov (United States)

    Saberi-Riseh, R; Javan-Nikkhah, M; Heidarian, R; Hosseini, S; Soleimani, P

    2004-01-01

    Wheat is an economic and important crop that provides approximately 20% of food calorie in the world. It is first crop in Iran and cultivated in the most areas of this country. Store-pit fungi make undesirable changes in quality and appearance of wheat grains. Even, some fungi produce different mycotoxins which are toxic to human and livestock's that use wheat grains as source of food. In this study, several samples were randomly collected from each of five store-pits located in different areas of Markazi Province including: Arak, Mahallat, Khomein, Saveh and Sarband. Grains were treated on PDA, and blotter, agar and washing test also used for isolating and detection of fungi. At least 100 grains per each sample were randomly used for each test and treatment. The fungi that determined in this study were Cochliobolus australiensis, Cladosporium herbarum, Epicoccum sp., Tilletia leavis, Aspergillus flavus, A. niger, A. fumigatus, Alternaria alternata, Alternaria sp., Penicillium italicum, P. digitatum, Fusarium sp., Rhizopus sp., Ustilago tritici, Scytalidium sp. Among these fungi the most isolates were belonged to Cladosporium, Alternaria, Rhizopus and Fusarium. Cladosporium herbarum was the most common in different sampling areas. Tilletia laevis and Ustilago tritici were just recovered in washing test. This study revealed that different fungi are associated with wheat grains in store-pits in Markazi Province. Some of them like Aspergillus flavus normally produce aflatoxin, a very toxic and carcinogenic mycotoxin that is harmful for human.

  9. Antimicrobial constituents from endophytic fungus Fusarium sp.

    Directory of Open Access Journals (Sweden)

    Hidayat Hussain

    2015-03-01

    Full Text Available Objective: To evaluate the antimicrobial potential of fraction of the fungus Fusarium sp. and study the tentative identification of their active constituents. Methods: Six compounds were purified from an fraction of endophytic fungus Fusarium sp. using column chromatography and their structures have been confirmed based on 1H and 13C nuclear magnetic resonance spectra, distortionless enhancement by polarization transfer, 2D COSY, heteronuclear multiple quantum correlation and heteronuclear multiple bond correlation experiments. The six isolated compounds were screened for antimicrobial activity using the agar well diffusion method. Results: Phytochemical investigation of endophytic fungus Fusarium sp. lead to the isolation and identification of the following compounds viz., colletorin B, colletochlorin B, LL-Z1272β (llicicolin B, 4,5-dihydroascochlorin, ascochlorin, and 4,5-dihydrodechloroascochlorin. Colletorin B and colletochlorin B displayed moderate herbicidal, antifungal and antibacterial activities towards Chlorella fusca, Ustilago violacea, Fusarium oxysporum, and Bacillus megaterium. On the other hand LL-Z1272β (llicicolin B showed moderate antifungal activity towards Ustilago violacea and Fusarium oxysporum while 4,5-dihydroascochlorin showed strong antibacterial activity towards Bacillus megaterium. Furthermore, 4,5-dihydrodechloroascochlorin showed very strong antifungal activity towards Eurotium repens. Conclusions: Antimicrobial activities demonstrated by five of the six isolated compounds clearly demonstrate that these fungi extracts and active compounds present a great potential for the food, cosmetic and pharmaceutical industries.

  10. A review on threat of gray leaf spot disease of maize in Asia

    Directory of Open Access Journals (Sweden)

    Narayan Bahadur Dhami

    2015-12-01

    Full Text Available Biotic and biotic constraints are yield limiting factors in maize producing regions. Among these gray leaf spot is a yield limiting foliar disease of maize in high land regions of Asia. This review is done from related different national and international journals, thesis, books, research papers etc. The objectives of this review are to become familiar with genetics and inheritance, epidemiology, symptoms and disease management strategies etc. High relative humidity, temperature, minimum tillage and maize monoculture are important factors responsible for disease development. The sibling species of Cercospora zeae-maydis (Tehon and Daniels, 1925 Group I and Group II and Cercospora sorghai var. maydis (Chupp, 1954 are associated with this disease. Pathogens colonize in maize debris. Conidia are the source of inoculums for disease spread. Severe blighting of leaves reduces sugars, stalk lodging and causes premature death of plants resulting in yield losses of up to 100%. Disease management through cultural practices is provisional. The use of fungicides for emergencies is effective however; their prohibitive cost and detrimental effects on the environment are negative consequences. The inheritance of tolerance is quantitative with small additive effects. The introgression of resistant genes among the crosses of resistant germplasm enhances the resistance. The crosses of resistant and susceptible germplasm possess greater stability than the crosses of susceptible and resistant germplasm. The development of gray leaf spot tolerant populations through tolerance breeding principle is an economical and sustainable approach to manage the disease.

  11. Mechanisms of Induced Resistance in Barley Against Drechslera teres.

    Science.gov (United States)

    Lyngs Jørgensen, H J; Lübeck, P S; Thordal-Christensen, H; de Neergaard, E; Smedegaard-Petersen, V

    1998-07-01

    ABSTRACT Quantitative and qualitative histopathological methods and molecular analyses were used to study the mechanisms by which preinoculation with either of the nonbarley pathogens, Bipolaris maydis and Septoria nodorum, inhibited growth of Drechslera teres. Collectively, our data suggest that induced resistance is the principal mechanism responsible for impeding the pathogen. The enhancement of resistance in the host was primarily manifested during penetration by D. teres, and after penetration, where growth of D. teres ceased soon after development of infection vesicles. Thus, 24 h after pretreatment with B. maydis or S. nodorum, the penetration frequency from D. teres appressoria was reduced from 42.7% in the controls to 9.5 and 14.8%, respectively. The reductions were associated with increased formation of fluorescent papillae in induced cells (early defense reaction). The postpenetrational inhibition of D. teres completely stopped fungal growth and was apparently linked to an enhancement of multicellular hypersensitive responses in inducer-treated leaves (late defense reaction). Papillae formation and multicellular hypersensitive reactions were also observed in fully susceptible, noninduced control leaves, but they were inadequate to stop fungal progress. Northern blots from leaves treated with either inducer alone support the conclusion that induced resistance is involved in suppression of D. teres by increased formation of papillae and hypersensitive reactions. Thus, the blots showed strong expression of several defense response genes that are involved in these reactions in barley attacked by Erysiphe graminis f. sp. hordei.

  12. Effect of Maize Hybrid and Foliar Fungicides on Yield Under Low Foliar Disease Severity Conditions.

    Science.gov (United States)

    Mallowa, Sally O; Esker, Paul D; Paul, Pierce A; Bradley, Carl A; Chapara, Venkata R; Conley, Shawn P; Robertson, Alison E

    2015-08-01

    Foliar fungicide use in the U.S. Corn Belt increased in the last decade; however, questions persist pertaining to its value and sustainability. Multistate field trials were established from 2010 to 2012 in Illinois, Iowa, Ohio, and Wisconsin to examine how hybrid and foliar fungicide influenced disease intensity and yield. The experimental design was in a split-split plot with main plots consisting of hybrids varying in resistance to gray leaf spot (caused by Cercospora zeae-maydis) and northern corn leaf blight (caused by Setosphaera turcica), subplots corresponding to four application timings of the fungicide pyraclostrobin, and sub-subplots represented by inoculations with either C. zeae-maydis, S. turcica, or both at two vegetative growth stages. Fungicide application (VT/R1) significantly reduced total disease severity relative to the control in five of eight site-years (Pfungicide-treated in three site-years. Fungicide application increased the yield by approximately 6% at Ohio in 2010, 5% at Wisconsin in 2010 and 6% in 2011. Yield differences ranged from 8,403 to 8,890 kg/ha in Wisconsin 2011 to 11,362 to 11,919 kg/ha in Wisconsin 2010. Results suggest susceptibility to disease and prevailing environment are important drivers of observed differences. Yield increases as a result of the physiological benefits of plant health benefits under low disease were not consistent.

  13. Enfermidades do sistema nervoso dos ruminantes no sul do Rio Grande do Sul Neurological diseases in ruminants in southern Brazil

    Directory of Open Access Journals (Sweden)

    Franklin Riet-Correa

    1998-06-01

    Full Text Available Descrevem-se os aspectos epidemiológicos, clínicos e patológicos das enfermidades do sistema nervoso central dos ruminantes, diagnosticadas na região Sul do Rio Grande do Sul, incluindo: abiotrofia cerebelar; hipoplasia cerebelar; hipermetria hereditária; artrogripose; hipomielinogênese congênita; abscesso cerebral; listeriose; tétano; botulismo; necrose simétrica focal; raiva; leucose; encefalite por herpesvírus bovino-5; febre catarral maligna; intoxicações por Solanum fastigiatum, Claviceps paspali, Ramaria flavo-brunnescens, Halimium brasiliense e Diplodia maydis; encefalopatia hepática causada por Senecio spp. e Echium plantagineum; cetose; coenurose; e síndrome espinhal.The main epidemiological, clinical and pathologic aspects of the diseases of the nervous system in cattle in Southern Rio Grande do Sul are described, including, the following conditions: cerebellar abiotrophy; cerebellar hypoplasia; congenital hypermetria; arthrogryposis; congenital hypomyelinogenesis; brain abscess; listeriose; tetanus; botulism; focal symmetrical encephalomalacia; rabies; leucosis; encephalitis by Herpesvirus Bovine-5; bovino malignant catarrh; intoxications by Solanum fastigiatum, Claviceps paspali, Halimium brasiliense, Diplodia maydis, and Ramaria flavo-brunnescens; hepatoencephalopaty caused by Senecio spp. and Echium plantagineum; ketosis; coenurosis; and spinal syndrome.

  14. AcEST: DK957206 [AcEST

    Lifescience Database Archive (English)

    Full Text Available _USTMA Putative DNA helicase INO80 OS=Ustilago ma... 59 2e-08 sp|Q0CA78|INO80_ASPTN Putative DNA helicase ino80... OS=Coccidioide... 55 3e-07 sp|A2R9H9|INO80_ASPNC Putative DNA helicase ino80 OS=Aspergillus... 54 4e-07 sp|Q872I5|INO80... OS=Magnaporthe... 54 8e-07 sp|Q2UTQ9|INO80_ASPOR Putative DNA helicase ino80... OS=Aspergillus... 53 1e-06 sp|A1CZE5|INO80_NEOFI Putative DNA helicase ino80 OS=Neosartorya... 53 1e-06 sp|Q4WTV7|INO80..._ASPFU Putative DNA helicase ino80 OS=Aspergillus... 53 1e-06 sp|A1C9W6|INO80_ASPCL Putative DNA helicase ino8

  15. Mycological and palynological studies of early medieval cultural layers from strongholds in Pszczew and Santok (western Poland

    Directory of Open Access Journals (Sweden)

    Kinga Mazurkiewicz-Zapałowicz

    2015-08-01

    Full Text Available Cultural layers from early medieval strongholds in Pszczew and Santok have been examined for the presence of pollen grains and spores as well as residues of fungi. The presence of the following remains has been recorded: fossil hyphopodia of Gaeumannomyces, teliospores of Puccinia, spores of Bipolaris, Thecaphora and Tilletia, teliospores of the genus Urocystis, Ustilago and Uromyces, ascocarps (perithecium of the Ascomycota or the pycnidium of Sphaeropsidales. A greater diversity and abundance of fungi spores sensu lato was recorded in Santok, as compared to Pszczew. Both early medieval sites recorded a significant proportion of cereal pollen, including Secale cereale. It remains an undisputed fact that the grains and other plants collected in both strongholds were strongly infected with fungi. The analysis of the cultural layers for the presence of fungi remains provides significant data on the presence of certain species of plants and their growth conditions in natural environments and in agriculture.

  16. Análisis de las reacciones de hipersensibilidad tipo I a los basidiomicetes en una población alérgica costarricense, durante 2009 Analysis of the reactions of type I hypersensitivity to the Basidiomycetes in a Costa Rican allergic population during 2009

    Directory of Open Access Journals (Sweden)

    Daniela Jaikel-Víquez

    2012-12-01

    Full Text Available Justificación y objetivos: Costa Rica es un país tropical en donde se ha encontrado una alta concentración de basidiosporas aéreas, las cuales miden alrededor de 4-13µm de diámetro, y son por definición aerobiológicas, capaces de penetrar a tracto respiratorio inferior, irrespetando la barrera nasal. El presente estudio tiene como objetivos encontrar posibles fuentes de contaminación ambiental por basidiomicetes y determinar si en Costa Rica sería necesario analizar, de forma rutinaria, la reactividad por alergenos a basidiomicetes en la población portadora de enfermedad rinobronquial alérgica. Métodos: se determinó el porcentaje de pacientes alérgicos a los extractos de basidiomicetes, tipo carbón Sporisorium cruentum y Ustilago cynodontis y tipo roya Puccinia graminis subsp. graminis. A la vez, se recolectaron panículas de Panicum maximum para evaluar su contaminación por basidiomicetes. Resultados: se encontró que el 59,23% de los pacientes eran alérgicos a hongos y que el tipo de hongo al que más pacientes presentaron alergia fueron los basidiomicetes. El 100% de las panículas recolectadas estaban contaminadas con un hongo del género Ustilago. Conclusión: se recomienda el testeo continuo por basidiomicetes, ya que estos son los hongos a los que la mayoría de los pacientes estudiados presentó alergia.Background and aim: Costa Rica is a tropical country in which a high concentration of aerial basidiospores has been found, which measure between 4-13 µm and are capable of arriving to the lower respiratory system, surpassing the nasal barrier. This study aims to find possible sources of environmental contamination by basidiomycetes and to determine if it is necessary to routinely analyze this allergen in the Costa Rican population that suffer from rhino bronchial allergy. Methods: The percentage of patients allergic to the extracts of basidiomycetes, specifically smuts like Sporisorium cruentum and Ustilago cynodontis

  17. 芦苇三种黑粉病菌冬孢子萌发试验初报

    Institute of Scientific and Technical Information of China (English)

    谭语词; 裴艳

    2007-01-01

    芦苇粒黑粉病(Ustilago phragmites Ling)、叶腥黑粉病(Tilletia nigrifaciens Langdon et Boughton)、穗腥黑粉病(Neovossia iowensis Hume et Hods)是发生在辽宁苇区芦苇上的三种黑粉病,国内外对该病菌的深入研究较少.我们通过冬孢子萌发试验证明,芦苇粒黑粉菌的冬孢子当年11月份已经成熟,在适当条件下即可萌发,而在山梨糖中的萌发率最高,在水中也有极个别孢子萌发,其它两种黑粉菌的冬孢子在糖液中较难萌发.

  18. A Survey of Phytopathogenic Fungi and Oomycetes in Riyadh, Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Arya Widyawan

    2015-06-01

    Full Text Available A survey of phytopathogenic fungi and Oomyceteswas conducted in Riyadh, Kingdom of Saudi Arabia duringOctober 2008 – May 2009. Total of 223 samples were collectedfrom four regions; Al-Kharj, Oyaynah, Old Diriyah, and Al Amariyah. Isolation was done using Potato Dextrose Agar(PDA. Infected parts were cut then sterilized in chlorox(10%, then were put in petridish that contain PDA andincubated at 25-27 °C. A total twelve genera of fungi andsingle genera of Oomycetes were isolated from the infectedplants and identifi ed as Fusarium spp., Alternaria spp.,Helmintosphorium (Bipolaris spp., Sclerotium spp., Rhizoctoniaspp., Cladosporium spp., Mauginiella scattae, Erysiphe spp.,Leveillula spp., Macrophomina phaseolina, Ustilago spp.,Ulocladium spp., and Phytium spp.

  19. 25S ribosomal RNA homologies of basidiomycetous yeasts: taxonomic and phylogenetic implications

    Science.gov (United States)

    Baharaeen, S.; Vishniac, H. S.

    1984-01-01

    Genera, families, and possibly orders of basidiomycetous yeasts can be defined by 25S rRNA homology and correlated phenotypic characters. The teleomorphic genera Filobasidium, Leucosporidium, and Rhodosporidium have greater than 96 relative binding percent (rb%) intrageneric 25S rRNA homology and significant intergeneric separation from each other and from Filobasidiella. The anamorphic genus Cryptococcus can be defined by morphology (monopolar budding), colony color, and greater than 75 rb% intrageneric homology; Vanrija is heterogeneous. Agaricostilbum (Phragmobasidiomycetes, Auriculariales), Hansenula (Ascomycotera, Endomycota), Tremella (Phragmobasidiomycetes, Tremellales), and Ustilago (Ustomycota, Ustilaginales) appear equally unrelated to the Cryptococcus, Filobasidiella, and Rhodosporidium spp. used as probes. The Filobasidiaceae and Sporidiaceae, Filobasidiales and Sporidiales, form coherent homology groups which appear to have undergone convergent 25S rRNA evolution, since their relatedness is much greater than that indicated by 5S rRNA homology. Ribosomal RNA homologies do not appear to measure evolutionary distance.

  20. AcEST: DK953287 [AcEST

    Lifescience Database Archive (English)

    Full Text Available elease 56.9) Link to BlastX Result : Swiss-Prot sp_hit_id P31622 Definition sp|P31622|GAG_JSRV Gag polyprotein OS=Sheep...ue sp|P31622|GAG_JSRV Gag polyprotein OS=Sheep pulmonary adenomatos... 36 0.17 sp|Q8YV57|Y2124_ANASP Unchara...in... 30 9.3 sp|Q4P0H7|BBP_USTMA Branchpoint-bridging protein OS=Ustilago may... 30 9.3 >sp|P31622|GAG_JSRV Gag polyprotein OS=Sheep...WR75_JSRV Gag polyprotein OS=Sheep pulmonary adenomatosis virus Align length 17 S...ue tr|Q9WR75|Q9WR75_JSRV Gag polyprotein OS=Sheep pulmonary adenoma... 36 2.0 tr|

  1. Análisis de las reacciones de hipersensibilidad tipo I a los basidiomicetes en una población alérgica costarricense, durante 2009

    Directory of Open Access Journals (Sweden)

    Daniela Jaikel-Víquez

    2012-12-01

    Full Text Available Justificación y objetivos: Costa Rica es un país tropical en donde se ha encontrado una alta concentración de basidiosporas aéreas, las cuales miden alrededor de 4-13µm de diámetro, y son por definición aerobiológicas, capaces de penetrar a tracto respiratorio inferior, irrespetando la barrera nasal. El presente estudio tiene como objetivos encontrar posibles fuentes de contaminación ambiental por basidiomicetes y determinar si en Costa Rica sería necesario analizar, de forma rutinaria, la reactividad por alergenos a basidiomicetes en la población portadora de enfermedad rinobronquial alérgica. Métodos: se determinó el porcentaje de pacientes alérgicos a los extractos de basidiomicetes, tipo carbón Sporisorium cruentum y Ustilago cynodontis y tipo roya Puccinia graminis subsp. graminis. A la vez, se recolectaron panículas de Panicum maximum para evaluar su contaminación por basidiomicetes. Resultados: se encontró que el 59,23% de los pacientes eran alérgicos a hongos y que el tipo de hongo al que más pacientes presentaron alergia fueron los basidiomicetes. El 100% de las panículas recolectadas estaban contaminadas con un hongo del género Ustilago. Conclusión: se recomienda el testeo continuo por basidiomicetes, ya que estos son los hongos a los que la mayoría de los pacientes estudiados presentó alergia.

  2. 新疆骆驼刺拮抗内生细菌XJAS-AB-13的分选·鉴定及其对玉米斑点病菌的生物防治潜能研究%Screening and Identification of An Antagonistic Endobacterium ( XJAS-AB-13 ) from Xinjiang Alhagi pseudalhagi Desv and Studies on Its Biocontrol Potentials to Maize Spot Pathogens

    Institute of Scientific and Technical Information of China (English)

    阿布都卡地尔·阿布力孜; 吾甫尔·米吉提; 阿依乃再·孜亚克; 阿力木江·阿合幼力

    2011-01-01

    [目的] 从骆驼刺(Alhagi pseudalhagi Desv)内生菌中筛选出对农作物致病菌具有较强生物活性的拮抗菌株,活体验证拮抗菌株的生物防治能力.[方法] 采用琼脂扩散法对常见的农作物致病菌做拮抗试验,结合传统病情统计和不同处理组玉米叶片中总蛋白含量以及防御酶活性测定活体验证拮抗菌株的生防能力.[结果] 筛选出对玉米大、小斑点病菌具有较强活性拮抗菌株XJAS-AB-13,根据拮抗内生细菌XJAS-AB-13的生理生化特征以及16S rDNA序列分析将其最终鉴定为枯草芽孢杆菌.该菌株发酵液对玉米大斑病菌和玉米小班病菌的抑菌圈直径分别达32.8和24.6 mm.活体试验结果显示,发酵液对玉米小斑病菌和玉米大斑病菌的防效率分别达到45.00%和63.33%.[结论] 拮抗菌株XJAS-AB-13在玉米大、小斑点病菌的生物防治中具有较好的应用潜力.%[ Objective ] The aim was to screen antagonistic endophytes from Alhagi pseudalhagi Desv to common crop pathogens, and verify biocontrol capacity of the active strains with in vivo experiment. [ Method] Agar diffusion method was used in screening the antagonistic endophytes. Traditional disease severity investigating method, measuring the total protein contents and defense enzymes activity of the maize leaf samples under the different treatment were used in testing the in vivo activity of the antagonistic strain. [Result] An antagonistic endobacterium (XJAS-AB-13) with strong antifungal activity to Exserohilum turcicum and Bipolaris maydis, was screened from Alhagi pseudalhagi Desv XJAS-AB-13 was finally identified as Bacillus subtilis according to physiological and biochemical characteristics and 16S rDNA sequence features. Inhibition zone diameter of fermentation broth of XJAS-AB-13 to Exserohilum turcicum and Bipolaris maydis reached 32. 8 and 24. 6 ram, respectively, in vivo experiment results showed that the biocontrol rate of XJAS-AB-13 to

  3. Pathogenic microorganisms of medicinal herbal drugs

    Directory of Open Access Journals (Sweden)

    Stević Tatjana

    2012-01-01

    Full Text Available All the parts of plants (root, leaf, flower naturally have a high level of microorganisms, bacteria and fungi, especially molds. Microbial contamination could be a result of inappropriate harvesting, cleaning of the raw plant material, unhygienic processing of the plants, unsuitable transport and storage. After examination of over 40 dried medicinal plant species, the lowest microbial quality was determined for Maydis stigma, Mentha leaf and herb, Equisetum herb, Calendula flower, Urtica leaf, Melissa leaf, Serpylli herb, Chamomilla flower etc. Although mixed infections are recorded with different types of fungus, Fusarium was observed as the most dominant genus in most of the tested drugs, followed by Aspergillus and Alternaria. In addition to these fungi species from the following genera were identified: Phoma, Cephalosporium, Nigrospora, Cladosporium, Epicoccum, Gliocladium, Myrothecium, Cercospora, Phomopsis, Verticillium, Dreschlera (=Bipolaris, Rhizoctonia, Septoria, Trichoderma, Curvularia, Stachybotrys, Trichothecium, Puccinia, Botrytis, Mucor and Rhizopus sp., depending on plant species.

  4. Incidência e severidade de doenças de quatro híbridos de milho cultivados com diferentes densidades de plantas Disease incidence and severity of four maize hybrids grown at different plant densities

    Directory of Open Access Journals (Sweden)

    Luís Sangoi

    2000-03-01

    Full Text Available Os híbridos modernos de milho apresentam maior potencial produtivo do que os cultivares utilizados no passado, especialmente sob situações de estresse, como densidade de semeadura elevada e deficiência hídrica. É possível que o maior rendimento de grãos dos genótipos modernos nessas condições se deva, em parte, a sua melhor sanidade. Este trabalho foi conduzido em Lages, SC, com o objetivo de avaliar a incidência e severidade das principais doenças foliares, de colmo e espiga de híbridos de milho cultivados no Planalto Catarinense, em diferentes densidades de semeadura. Utilizou-se o delineamento experimental de blocos ao acaso com parcelas subdivididas. Na parcela principal, foram testadas três populações de planta, equivalentes a 50000, 75000 e 100000pl ha-1. Nas subparcelas, testaram-se quatro híbridos: Ag 12, Ag 28, Ag 303 e Ag 9012, os quais foram cultivados de forma expressiva nas décadas de 60, 70, 80, e 90, respectivamente. O aumento da população de plantas promoveu um incremento na incidência das doenças de colmo causadas por Fusarium moniliforme, Colletotrichum graminicola e Diplodia maydis. Esse aumento na quantidade de doenças foi mais intenso nos híbridos Ag 12 e Ag 28, contribuindo para aumentar a percentagem de plantas acamadas e quebradas destes materiais na colheita. O híbrido Ag 9012 apresentou menor incidência e severidade das doenças de espiga causadas por Fusarium moniliforme e Diplodia maydis, independentemente da densidade de semeadura. Não foi constatada menor severidade a doenças foliares no híbrido Ag 9012, em relação aos híbridos mais antigos.Modern maize hybrids have greater yield potential than cultivars used in the past, particularly under stressful conditions such as high plant population and water deficit. It is possible that the yield advantage presented by modern genotypes is partially due to their better ability to resist major maize diseases. This experiment was carried out to

  5. Studies on the Antimicrobial Activity of Chitooligosaccharides from Housefly Larvae,Musca domestica vicina Macquart (Diptera:Muscidae)

    Institute of Scientific and Technical Information of China (English)

    WEI Xin-kui; LEI Chao-liang

    2004-01-01

    The inhibition effect of chitooligosaccharides from housefly larvae on pathogens ofcrops seeds, fruits or vegetables was studied and the main factors influencing theantimicrobial activity of chitooligosaccharides were also investigated. It was foundthat chitooligosaccharides from housefly larvae had wide spectrum fungistasis. It canstrongly inhibit the development of 31 kinds of plant pathogenic fungi such as corticiumrolfsii Saccardo. The results indicated that the degree of deacelylation (DD) or theaverage molecular weight (MW) of chitooligosaccharides were related to the antimicrobialactivity. The antimicrobial activity increased with the rising of DD or the declining ofMW. Pot culture results showed that chitooligosaccharide could enhance the rate ofgermination and emergence of the seeds of maize, wheat and cotton. Chitooligosaccharidehad certain effect on corn southern leaf blight caused by Helminthosporium maydis.

  6. Design, synthesis, anti-TMV, fungicidal, and insecticidal activity evaluation of 1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid derivatives based on virus inhibitors of plant sources.

    Science.gov (United States)

    Song, Hong-jian; Liu, Yong-xian; Liu, Yu-xiu; Huang, Yuan-qiong; Li, Yong-qiang; Wang, Qing-min

    2014-11-15

    By drawing the creation ideas of botanical pesticides, a series of tetrahydro-β-carboline-3-carboxylic acid derivatives were designed and synthesized, and first evaluated for their anti-TMV, fungicidal and insecticidal activities. Most of these derivatives exhibited good antiviral activity against TMV both in vitro and in vivo. Especially, the activities of compounds 8 and 15 in vivo were higher than that of ribavirin. The compound 8 exhibited more than 70% fungicidal activities against Cercospora arachidicola Hori, Alternaria solani, Bipolaris maydis, and Rhizoctonia solani at 50mg/kg, compounds 16 and 20 exhibited more than 60% insecticidal activities against Mythimna separate and Ostrinia nubilalis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. 河西走廊制种玉米生育中后期叶部真菌病害诊断与病原初步鉴定%Diagnosis and Preliminary Identification on Pathogeny of Foliage Fungal Diseases at Medium-late Stage of Maize in Hexi Corridor

    Institute of Scientific and Technical Information of China (English)

    陈广泉; 闫治斌; 陈占伟; 郑天翔; 邢会琴; 张建超; 费永祥

    2015-01-01

    对河西走廊制种玉米生育中后期叶部真菌病害进行田间调查,并将采集到的标本进行组织分离、病原鉴定和致病性测定.结果表明,引起制种玉米生育中后期叶部真菌病害的植物病原有13种,其中由Curvulairia lunata(Wakke) Boed病原引起的玉米弯孢菌叶斑病是玉米叶部的一种新病害,在田间发病率较高,病情指数较大,危害十分严重;由Exserohilum turcicum(Pass.) Leonard et Suggs和Bipolaris maydis(Nishik.et Miyabe) Shoemaker病原引起的大斑病和小斑病是玉米生育中后期叶部发生的主要真菌病害;Physoderma maydis Miyabe和Fusarium subglutinans(Wollenwe.et Reinking)P.E.Nelson et al病原引起的褐斑病和顶腐病有加重发生的趋势;Puccinia sorghi Schw引起的普通锈病在局部区域的一些品种和组合上发病较为严重;其他叶部病害零星发生,对制种玉米正常的生长发育影响较小,一般不会造成流行为害.

  8. Genome and transcriptome analysis of the basidiomycetous yeast Pseudozyma antarctica producing extracellular glycolipids, mannosylerythritol lipids.

    Directory of Open Access Journals (Sweden)

    Tomotake Morita

    Full Text Available Pseudozyma antarctica is a non-pathogenic phyllosphere yeast known as an excellent producer of mannosylerythritol lipids (MELs, multi-functional extracellular glycolipids, from vegetable oils. To clarify the genetic characteristics of P. antarctica, we analyzed the 18 Mb genome of P. antarctica T-34. On the basis of KOG analysis, the number of genes (219 genes categorized into lipid transport and metabolism classification in P. antarctica was one and a half times larger than that of yeast Saccharomyces cerevisiae (140 genes. The gene encoding an ATP/citrate lyase (ACL related to acetyl-CoA synthesis conserved in oleaginous strains was found in P. antarctica genome: the single ACL gene possesses the four domains identical to that of the human gene, whereas the other oleaginous ascomycetous species have the two genes covering the four domains. P. antarctica genome exhibited a remarkable degree of synteny to U. maydis genome, however, the comparison of the gene expression profiles under the culture on the two carbon sources, glucose and soybean oil, by the DNA microarray method revealed that transcriptomes between the two species were significantly different. In P. antarctica, expression of the gene sets relating fatty acid metabolism were markedly up-regulated under the oily conditions compared with glucose. Additionally, MEL biosynthesis cluster of P. antarctica was highly expressed regardless of the carbon source as compared to U. maydis. These results strongly indicate that P. antarctica has an oleaginous nature which is relevant to its non-pathogenic and MEL-overproducing characteristics. The analysis and dataset contribute to stimulate the development of improved strains with customized properties for high yield production of functional bio-based materials.

  9. Efeito de época de semeadura e adubação na mancha-foliar de Phaeosphaeria em milho

    Directory of Open Access Journals (Sweden)

    Pegoraro Diego Girardi

    2001-01-01

    Full Text Available O aumento da área de cultivo de milho, aliado a algumas práticas culturais, tem favorecido o surgimento da mancha-foliar causada pelo fungo Phaeosphaeria maydis. Como conseqüência, o rendimento de grãos tem decrescido, especialmente em áreas com plantio direto e maior amplitude da época de semeadura. O trabalho teve como objetivo avaliar a severidade de P. maydis em seis híbridos de milho, em cinco épocas de semeadura e em dois níveis de adubação. O experimento foi conduzido em Xanxerê, SC, na safra 1997/98, em blocos casualizados, com parcelas subdivididas, em duas repetições. As plantas foram avaliadas 30 dias após o florescimento, e a porcentagem da área foliar afetada foi registrada. Na medida em que a semeadura do milho foi retardada, houve um aumento na severidade da moléstia e uma redução no rendimento de grãos. As doses de adubação não apresentaram diferença quanto à severidade da moléstia. O mês de setembro apresentou menor severidade da mancha-foliar e maior rendimento de grãos. Houve uma correlação significativa entre o rendimento de grãos e a severidade da moléstia.

  10. Microflora fúngica de sementes de milho em ambientes de armazenamento

    Directory of Open Access Journals (Sweden)

    Tanaka Maria Aparecida de Souza

    2001-01-01

    Full Text Available Durante o armazenamento, vários fungos podem permanecer associados às sementes de milho, causando deterioração ou se mantendo viáveis, infectando posteriormente a plântula. Objetivou-se neste trabalho verificar a sobrevivência de fungos associados às sementes de milho durante 12 meses de armazenamento, em câmara fria (14ºC; 40% UR e em ambiente não controlado. Observaram-se com maior freqüência os fungos de campo Alternaria alternata, Bipolaris maydis, Cephalosporium acremonium, Cladosporium herbarum, Fusarium moniliforme e Rhizoctonia solani, além de Rhizopus spp. e Trichoderma spp., cuja sobrevivência decresceu ao longo do armazenamento, de modo muito mais acentuado em condição de ambiente não controlado, em comparação à câmara fria. Aspergillus e Penicillium tiveram suas incidências aumentadas ao longo do período, principalmente em ambiente não controlado. Nessa condição, a sobrevivência de Fusarium moniliforme foi reduzida gradativamente até o final dos doze meses; comparativamente, em câmara fria, a viabilidade do fungo foi menos afetada. Bipolaris maydis manteve-se viável na maioria dos lotes, durante todo o armazenamento em câmara fria; em ambiente não controlado, o fungo sobreviveu durante quatro a dez meses, dependendo do lote avaliado. O armazenamento em ambiente não controlado, embora tenha provocado a redução do inóculo de F. moniliforme e outros fungos importantes, poderia acelerar o processo de deterioração das sementes. Em câmara fria, por outro lado, a viabilidade dos fungos é favorecida, comprometendo a qualidade sanitária das sementes.

  11. FORMAS DE PENETRAÇÃO DO GÊNERO CERCOSPORA

    Directory of Open Access Journals (Sweden)

    PAULA, Paulo Victor Augusto Azevedo de

    2015-11-01

    Full Text Available The genus Cercospora has species that cause diseases in important crops causing yield losses worldwide. These pathogens are commonly associated with leaf spot symptoms, but can cause injury of flowers, fruits, bracts, pedicels and seeds in various plant families. This review aims to describe the different forms of penetration of the genus Cercospora, causing diseases in crops of commercial importance, such as C. beticola in sugar beet, cassava in C. heningsii, C. coffeicola in coffee, C. sojina in soybean, C. zeae-maydis in corn, C. ricinella in castor and C. kikuchii in soybean. The penetration may occur through the stomata of the leaf of the host can also occur through the epidermal tissue injury and which occur by physical force and/or the action of enzymes directly through the epidermis of the host body. And for the development of the germ tube, different species may develop one or more germ tubes. Then, Cercospora spp has different ways of penetration depending of your host. O gênero Cercospora destaca-se por ser o agente etiológico de doenças em importantes culturas acarretando perdas de produção em todo o mundo. Esses patógenos comumente estão associados a sintomas de manchas foliares, mas podem causar lesões em flores, frutos, brácteas, sementes e pedicelos em diversas famílias de plantas. Essa revisão tem como objetivo descrever as diferentes formas de penetração de espécies do gênero Cercospora, agentes etiológicos de doenças em culturas de grande importância comercial, como C. beticola em beterraba, C. heningsii em mandioca, C. coffeicola em cafeeiro, C. sojina em soja, C. zeae-maydis em milho, C. ricinella em mamona e C. kikuchi em soja. A penetração pode ocorrer através dos estômatos das folhas dos hospedeiros, também pode ocorrer através de ferimentos nos tecidos da epiderme e ainda por força física e/ou ação de enzimas diretamente através da epiderme do órgão do hospedeiro. E em relação ao tubo

  12. Effects of Bacillus amyloliquefaciens B9601-Y2 on Diseases Control and Growth-promotion of Maize%解淀粉芽孢杆菌B9601-Y2对玉米的防病促生长效果研究

    Institute of Scientific and Technical Information of China (English)

    崔文艳; 何朋杰; 尚娟; 李艳云; 吴毅歆; 何月秋

    2015-01-01

    By dual-culture confrontation assay(DCA) and pot culture test, to study the effect of Bacillus amylo?liquefaciens B9601-Y2(Y2) to control southern leaf blight(SLB) and stalk rot and increase biomass of maize. DCA showed that formulate suspension(FS), cell-free filtrate(CF) and cell suspension(CS) of Y 2 could inhibit mycelial growth of Bipolaris maydis and Fusarium verticillioides. Pot culture test indicated that those suspensions could con?trol over 60%of SLB and over 50%of stalk rot with the tendency FS>CS>CF. Those suspensions promoted over 20%of root length, plant type height, dry weight of above-ground and dry weight of under-ground for healthy maize and for the disease seedlings inoculated with B. maydis and F. verticillioides by spray or drenching. Therefore, Y2 has the potential serving as a root and leaf biofertilizer with disease control effect for maize.%通过平板对峙培养及盆栽试验,研究解淀粉芽孢杆菌B9601-Y2(Y2)防治玉米小斑病和茎基腐病以及提高玉米生物量的效果.平板对峙培养结果表明,Y2发酵液、无菌滤液及菌体悬液对玉米小斑病菌和茎基腐病原菌均有拮抗作用.盆栽实验结果显示,3种处理对玉米小斑病防效60%左右,对茎腐病防效50%左右,防效依次为Y2发酵液>菌体悬液>无菌滤液.单独使用Y2发酵液、无菌滤液及菌体悬液处理的苗根长、株高、鲜重和干重均显著高于清水和病原菌对照20%以上,喷施或灌溉接种病原菌的玉米苗生长量显著高于单独接种病原菌处理,有效消除了病原菌对植株生长的影响.因此,生防菌Y2具有用于防治玉米病害和具有根肥及叶面肥的潜力.

  13. Desfolha, população de plantas e precocidade do milho afetam a incidência e a severidade de podridões de colmo Defoliation, plant population, and earliness of maize affect the incidence and severity of stalk rots

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo Bassay Blum

    2003-10-01

    Full Text Available As relações entre fonte e dreno interferem sobre a ocorrência de podridões de colmo na cultura do milho (Zea mays. O estudo foi conduzido em Lages, SC, para avaliar o efeito da desfolha, população de plantas e híbridos sobre as podridões do colmo do milho. O delineamento experimental foi de blocos ao acaso com parcelas sub-subdivididas e três repetições. Os híbridos P32R21 (superprecoce, Premium (precoce e C333B (tardio foram testados sem desfolha e com 50% de desfolha no estágio de espigamento nas populações de 25.000, 50.000, 75.000 e 100.000 plantas ha-1. Avaliaram-se a incidência e a severidade das podridões do colmo causadas por Colletotrichum graminicola, Diplodia maydis e Fusarium moniliforme. O híbrido P32R21 foi o mais afetado pelos fungos causadores de podridões. Nos híbridos P32R21 e Premium, as plantas desfolhadas foram mais severamente infectadas pelos patógenos do que as intactas. O aumento da população de plantas aumentou a intensidade (incidência ou severidade de podridões de colmo nas plantas intactas e diminuiu a ocorrência nas desfolhadas.The relationship between source and sink interferes on maize (Zea mays stalk rot occurrence. This study was conducted in Lages, SC, Brazil, to evaluate the effect of defoliation, plant population, and hybrids on the intensity (incidence or severity of maize stalk rots. The experiment was set in a randomized complete block design (three replications with a split-split-plot arrangement. The hybrids P32R21 (very early, Premium (early and C333B (late were tested without defoliation and with 50% defoliation on ear formation stage at plant populations of 25,000, 50,000, 75,000, and 100,000 plants ha-1. The incidence and severity of Colletotrichum graminicola, Diplodia maydis and Fusarium moniliforme stalk rots were evaluated. The hybrid P32R21 was the most affected by stalk rots. Defoliation enhanced rots on P32R21 and Premium. Enhancements in plant population increased

  14. Resistance to gray leaf spot of maize: genetic architecture and mechanisms elucidated through nested association mapping and near-isogenic line analysis.

    Directory of Open Access Journals (Sweden)

    Jacqueline M Benson

    2015-03-01

    Full Text Available Gray leaf spot (GLS, caused by Cercospora zeae-maydis and Cercospora zeina, is one of the most important diseases of maize worldwide. The pathogen has a necrotrophic lifestyle and no major genes are known for GLS. Quantitative resistance, although poorly understood, is important for GLS management. We used genetic mapping to refine understanding of the genetic architecture of GLS resistance and to develop hypotheses regarding the mechanisms underlying quantitative disease resistance (QDR loci. Nested association mapping (NAM was used to identify 16 quantitative trait loci (QTL for QDR to GLS, including seven novel QTL, each of which demonstrated allelic series with significant effects above and below the magnitude of the B73 reference allele. Alleles at three QTL, qGLS1.04, qGLS2.09, and qGLS4.05, conferred disease reductions of greater than 10%. Interactions between loci were detected for three pairs of loci, including an interaction between iqGLS4.05 and qGLS7.03. Near-isogenic lines (NILs were developed to confirm and fine-map three of the 16 QTL, and to develop hypotheses regarding mechanisms of resistance. qGLS1.04 was fine-mapped from an interval of 27.0 Mb to two intervals of 6.5 Mb and 5.2 Mb, consistent with the hypothesis that multiple genes underlie highly significant QTL identified by NAM. qGLS2.09, which was also associated with maturity (days to anthesis and with resistance to southern leaf blight, was narrowed to a 4-Mb interval. The distance between major leaf veins was strongly associated with resistance to GLS at qGLS4.05. NILs for qGLS1.04 were treated with the C. zeae-maydis toxin cercosporin to test the role of host-specific toxin in QDR. Cercosporin exposure increased expression of a putative flavin-monooxygenase (FMO gene, a candidate detoxification-related gene underlying qGLS1.04. This integrated approach to confirming QTL and characterizing the potential underlying mechanisms advances the understanding of QDR and will

  15. Synthesis and Anti-fungal Activity of Chalcones with Pyridyls%吡啶基查尔酮类化合物合成及其抑菌活性

    Institute of Scientific and Technical Information of China (English)

    詹帅军; 金洪; 陶科; 侯太平

    2016-01-01

    为了发现高效、低毒的农用先导化合物,本文采用活性亚结构拼接原理,设计并合成了一类含吡啶基查尔酮类化合物。初步抑菌活性测试结果表明,在测试浓度为100μg/mL时,大多数化合物对供试的水稻纹枯病菌(Rhizoctorzia solani)、苹果轮纹病(Physolospora piricola)、小麦赤霉病菌(Fusarium graminearum)、玉米小斑病菌(Bipolaris maydis)四种植物病原菌均具有一定的抑菌活性,其中化合物I9,I15和I16对水稻纹枯病菌的抑制率在90%以上。病原菌毒力测试表明,化合物I16的EC50值为35.43μg/mL,高于对照药多菌灵(40.56μg/mL)的值,此外,化合物I16还表现出了较宽的抑菌谱,其对四种植物病原菌的抑菌率均大于80%。%In order to find new anti-fungal leading compounds with high performance and low toxicity, this paper designed and synthesized a series of chalcone analogs with pyridyls according to a principle of splice. Bioassays showed that most of target compounds had a certain anti-fungal activities against Rhizoctorzia solani, Physalospora piricola, Fusarium graminearum and Bipolaris maydis at a dosage of 100μg/mL. Inhibition rates of compounds I9, I15 and I16 were more than 90%. The EC50 value of compound I16 was 35.43 μg/mL, which was higher than the control group Carbendazim(40.56μg/mL). Furthermore, compound I16 showed a broad-spectrum fungicidal activity, its fungicidal rate against four kinds of fungi were greater than 80%.

  16. The effects of meteorological factors on airborne fungal spore concentration in two areas differing in urbanisation level

    Science.gov (United States)

    Oliveira, M.; Ribeiro, H.; Delgado, J. L.; Abreu, I.

    2009-01-01

    Although fungal spores are an ever-present component of the atmosphere throughout the year, their concentration oscillates widely. This work aims to establish correlations between fungal spore concentrations in Porto and Amares and meteorological data. The seasonal distribution of fungal spores was studied continuously (2005-2007) using volumetric spore traps. To determine the effect of meteorological factors (temperature, relative humidity and rainfall) on spore concentration, the Spearman rank correlation test was used. In both locations, the most abundant fungal spores were Cladosporium, Agaricus, Agrocybe, Alternaria and Aspergillus/Penicillium, the highest concentrations being found during summer and autumn. In the present study, with the exception of Coprinus and Pleospora, spore concentrations were higher in the rural area than in the urban location. Among the selected spore types, spring-autumn spores ( Coprinus, Didymella, Leptosphaeria and Pleospora) exhibited negative correlations with temperature and positive correlations both with relative humidity and rainfall level. On the contrary, late spring-early summer (Smuts) and summer spores ( Alternaria, Cladosporium, Epicoccum, Ganoderma, Stemphylium and Ustilago) exhibited positive correlations with temperature and negative correlations both with relative humidity and rainfall level. Rust, a frequent spore type during summer, had a positive correlation with temperature. Aspergillus/Penicillium, showed no correlation with the meteorological factors analysed. This knowledge can be useful for agriculture, allowing more efficient and reliable application of pesticides, and for human health, by improving the diagnosis and treatment of respiratory allergic disease.

  17. A RALDH-like enzyme involved in Fusarium verticillioides development.

    Science.gov (United States)

    Díaz-Sánchez, Violeta; Limón, M Carmen; Schaub, Patrick; Al-Babili, Salim; Avalos, Javier

    2016-01-01

    Retinaldehyde dehydrogenases (RALDHs) convert retinal to retinoic acid, an important chordate morphogen. Retinal also occurs in some fungi, such as Fusarium and Ustilago spp., evidenced by the presence of rhodopsins and β-carotene cleaving, retinal-forming dioxygenases. Based on the assumption that retinoic acid may also be formed in fungi, we searched the Fusarium protein databases for RALDHs homologs, focusing on Fusarium verticillioides. Using crude lysates of Escherichia coli cells expressing the corresponding cDNAs, we checked the capability of best matches to convert retinal into retinoic acid in vitro. Thereby, we identified an aldehyde dehydrogenase, termed CarY, as a retinoic acid-forming enzyme, an activity that was also exerted by purified CarY. Targeted mutation of the carY gene in F. verticillioides resulted in alterations of mycelia development and conidia morphology in agar cultures, and reduced capacity to produce perithecia as a female in sexual crosses. Complementation of the mutant with a wild-type carY allele demonstrated that these alterations are caused by the lackof CarY. However, retinoic acid could not be detected by LC-MS analysis either in the wild type or the complemented carY strain in vivo, making elusive the connection between CarY enzymatic activity and retinoic acid formation in the fungus.

  18. Exploration and Swiss field-testing of a viral gene for specific quantitative resistance against smuts and bunts in wheat.

    Science.gov (United States)

    Schlaich, Thomas; Urbaniak, Bartosz; Plissonnier, Marie-Laure; Malgras, Nicole; Sautter, Christof

    2007-01-01

    The viral gene for the killer protein 4 (KP4) has been explored for its antifungal effect in genetically modified wheat to defeat specifically the seed-transmitted smut and bunt diseases. In vitro both important seed-transmitted diseases of wheat, loose smut (Ustilago tritici) and stinking smut (Tilletia caries), are susceptible to KP4, whereas all other organisms tested so far proved to be not susceptible to KP4. For studies in planta we used stinking smut as a model fungus. In greenhouse experiments, two KP4-transgenic wheat lines showed up to 30% lower symptom development as compared to the nontransgenic control. As the last step in the proof of concept, field-testing has shown for the first time increased fungal resistance of a transgene in wheat. Due to its specificity against smuts and bunts, KP4 presents a very low risk to humans and the environment. Field-testing in Switzerland is regulated by a strong law, which for research is acceptable if legally and scientifically correctly applied.

  19. Bioprospecting and evolving alternative xylose and arabinose pathway enzymes for use in Saccharomyces cerevisiae.

    Science.gov (United States)

    Lee, Sun-Mi; Jellison, Taylor; Alper, Hal S

    2016-03-01

    Bioprospecting is an effective way to find novel enzymes from strains with desirable phenotypes. Such bioprospecting has enabled organisms such as Saccharomyces cerevisiae to utilize nonnative pentose sugars. Yet, the efficiency of this pentose catabolism (especially for the case of arabinose) remains suboptimal. Thus, further pathway optimization or identification of novel, optimal pathways is needed. Previously, we identified a novel set of xylan catabolic pathway enzymes from a superior pentose-utilizing strain of Ustilago bevomyces. These enzymes were used to successfully engineer a xylan-utilizing S. cerevisiae through a blended approach of bioprospecting and evolutionary engineering. Here, we expanded this approach to xylose and arabinose catabolic pathway engineering and demonstrated that bioprospected xylose and arabinose catabolic pathways from U. bevomyces offer alternative choices for enabling efficient pentose catabolism in S. cerevisiae. By introducing a novel set of xylose catabolic genes from U. bevomyces, growth rates were improved up to 85 % over a set of traditional Scheffersomyces stipitis pathway genes. In addition, we suggested an alternative arabinose catabolic pathway which, after directed evolution and pathway engineering, enabled S. cerevisiae to grow on arabinose as a sole carbon source in minimal medium with growth rates upwards of 0.05 h(-1). This pathway represents the most efficient growth of yeast on pure arabinose minimal medium. These pathways provide great starting points for further strain development and demonstrate the utility of bioprospecting from U. bevomyces.

  20. Xylan catabolism is improved by blending bioprospecting and metabolic pathway engineering in Saccharomyces cerevisiae.

    Science.gov (United States)

    Lee, Sun-Mi; Jellison, Taylor; Alper, Hal S

    2015-04-01

    Complete utilization of all available carbon sources in lignocellulosic biomass still remains a challenge in engineering Saccharomyces cerevisiae. Even with efficient heterologous xylose catabolic pathways, S. cerevisiae is unable to utilize xylose in lignocellulosic biomass unless xylan is depolymerized to xylose. Here we demonstrate that a blended bioprospecting approach along with pathway engineering and evolutionary engineering can be used to improve xylan catabolism in S. cerevisiae. Specifically, we perform whole genome sequencing-based bioprospecting of a strain with remarkable pentose catabolic potential that we isolated and named Ustilago bevomyces. The heterologous expression of xylan catabolic genes enabled S. cerevisiae to grow on xylan as a single carbon source in minimal medium. A combination of bioprospecting and metabolic pathway evolution demonstrated that the xylan catabolic pathway could be further improved. Ultimately, engineering efforts were able to achieve xylan conversion into ethanol of up to 0.22 g/L on minimal medium compositions with xylan. This pathway provides a novel starting point for improving lignocellulosic conversion by yeast.

  1. Estimating the abundance of airborne pollen and fungal spores at variable elevations using an aircraft: how high can they fly?

    Science.gov (United States)

    Damialis, Athanasios; Kaimakamis, Evangelos; Konoglou, Maria; Akritidis, Ioannis; Traidl-Hoffmann, Claudia; Gioulekas, Dimitrios

    2017-03-01

    Airborne pollen and fungal spores are monitored mainly in highly populated, urban environments, for allergy prevention purposes. However, their sources can frequently be located outside cities’ fringes with more vegetation. So as to shed light to this paradox, we investigated the diversity and abundance of airborne pollen and fungal spores at various environmental regimes. We monitored pollen and spores using an aircraft and a car, at elevations from sea level to 2,000 m above ground, in the region of Thesssaloniki, Greece. We found a total of 24 pollen types and more than 15 spore types. Pollen and spores were detected throughout the elevational transect. Lower elevations exhibited higher pollen concentrations in only half of plant taxa and higher fungal spore concentrations in only Ustilago. Pinaceae and Quercus pollen were the most abundant recorded by airplane (>54% of the total). Poaceae pollen were the most abundant via car measurements (>77% of the total). Cladosporium and Alternaria spores were the most abundant in all cases (aircraft: >69% and >17%, car: >45% and >27%, respectively). We conclude that pollen and fungal spores can be diverse and abundant even outside the main source area, evidently because of long-distance transport incidents.

  2. Airborne Spore Analysis of Karabük Atmosphere

    Directory of Open Access Journals (Sweden)

    Ayşe Kaplan

    2014-05-01

    Full Text Available In order to identify types and amounts of airborne allergenic spore dispersal in the atmosphere of Karabük by gravimetric method in 2006 and 2007, two Durham samplers were situated on roof and garden of Technical Education Faculty of Karabük University between the dates January 1, 2006 and December 31, 2007. As a result of the analysis a total of 2822.2±625.01 spore/cm2 spore quantity belonging to 21 types was identified. Of this total, 1106±250.33 spore/cm² was observed in 2006 and 1716±374.68 spore/cm² was observed in 2007. Spore concentrations revealed no statistically differences between two samplers (t=0.1527-1.1355, p>0.05. The relationship between spore concentrations and meteorological factors was displayed by Spearman Correlation analysis. The highest quantity of fungal spores and Myxomycetes were determined in June and July. Cladosporium, Alternaria, Ustilago, Myxomycetes and unidentified Ascomycetes spores were recorded as dominant. In the end of this study, a two-year spore calendar was prepared.

  3. Investigation and Identification of Sugarcane Diseases in Guangxi Province%广西甘蔗病害种类调查与鉴定

    Institute of Scientific and Technical Information of China (English)

    霍秀娟; 李朝生; 陆荣生

    2013-01-01

    2010~2012年采用定点踏查、普查和定时系统调查的方式对广西甘蔗病害种类进行了调查.结果表明,共鉴定出甘蔗病害21种,其中真菌病害15种,细菌病害1种,病毒病害2种,线虫病害3种;甘蔗鞭黑穗病、凤梨病、梢腐病、宿根矮化病和花叶病发生较普遍,是广西蔗区的主要病害.%An investigation on sugarcane diseases in Guangxi was conducted during 2010-2012 by on-the-spot survey,general survey,system investigation at fixed spot and time.The results showed that there are 21 species of sugarcane diseases in Guangxi Province,including 15 kinds of fungal diseases,1 kind of bacterial disease,2 kinds of virus diseases and 3 kinds of nematodes diseases.Ustilago scitaminea Sydow,Ceratocystis paradoxa (Dode)Moreau,Fusarium moniliforme Sheldeon,Clavibacter xyli subsp.Xyli Davis etc and Sugarcane mosaic virus were widely spread.

  4. A RALDH-like enzyme involved in Fusarium verticillioides development

    KAUST Repository

    Díaz-Sánchez, Violeta

    2015-12-11

    Retinaldehyde dehydrogenases (RALDHs) convert retinal to retinoic acid, an important chordate morphogen. Retinal also occurs in some fungi, such as Fusarium and Ustilago spp., evidenced by the presence of rhodopsins and β–carotene cleaving, retinal-forming dioxygenases. Based on the assumption that retinoic acid may also be formed in fungi, we searched the Fusarium protein databases for RALDHs homologs, focusing on Fusarium verticillioides. Using crude lysates of Escherichia coli cells expressing the corresponding cDNAs, we checked the capability of best matches to convert retinal into retinoic acid in vitro. Thereby, we identified an aldehyde dehydrogenase, termed CarY, as a retinoic acid-forming enzyme, an activity that was also exerted by purified CarY. Targeted mutation of the carY gene in F. verticillioides resulted in alterations of mycelia development and conidia morphology in agar cultures, and reduced capacity to produce perithecia as a female in sexual crosses. Complementation of the mutant with a wild-type carY allele demonstrated that these alterations are caused by the lack of CarY. However, retinoic acid could not be detected by LC-MS analysis either in the wild type or the complemented carY strain in vivo, making elusive the connection between CarY enzymatic activity and retinoic acid formation in the fungus.

  5. A circadian rhythm regulating hyphal melanization in Cercospora kikuchii.

    Science.gov (United States)

    Bluhm, Burton H; Burnham, A Michele; Dunkle, Larry D

    2010-01-01

    Many metabolic and developmental processes in fungi are controlled by biological rhythms. Circadian rhythms approximate a daily (24 h) cycle and have been thoroughly studied in the model fungus, Neurospora crassa. However relatively few examples of true circadian rhythms have been documented among other filamentous fungi. In this study we describe a circadian rhythm underlying hyphal melanization in Cercospora kikuchii, an important pathogen of soybean. After growth in light or light : dark cycles, colonies transferred to darkness produced zonate bands of melanized hyphae interspersed with bands of hyaline hyphae. Rhythmic production of bands was remarkably persistent in the absence of external cues, lasting at least 7 d after transfer to darkness, and was compensated over a range of temperatures. As in N. crassa, blue light but not red light was sufficient to entrain the circadian rhythm in C. kikuchii, and a putative ortholog of white collar-1, one of the genes required for light responses in N. crassa, was identified in C. kikuchii. Circadian regulation of melanization is conserved in other members of the genus: Similar rhythms were identified in another field isolate of C. kikuchii as well as field isolates of C. beticola and C. sorghi, but not in wild-type strains of C. zeae-maydis or C. zeina. This report represents the first documented circadian rhythm among Dothideomycete fungi and provides a new opportunity to dissect the molecular basis of circadian rhythms among filamentous fungi.

  6. Comparison of fungi within the Gaeumannomyces-Phialophora complex by analysis of ribosomal DNA sequences.

    Science.gov (United States)

    Bryan, G T; Daniels, M J; Osbourn, A E

    1995-02-01

    Four ascomycete species of the genus Gaeumannomyces infect roots of monocotyledons. Gaeumannomyces graminis contains four varieties, var. tritici, var. avenae, var. graminis, and var. maydis. G. graminis varieties tritici, avenae, and graminis have Phialophora-like anamorphs and, together with the other Gaeumannomyces and Phialophora species found on cereal roots, constitute the Gaeumannomyces-Phialophora complex. Relatedness of a number of Gaeumannomyces and Phialophora isolates was assessed by comparison of DNA sequences of the 18S rRNA gene, the 5.8S rRNA gene, and the internal transcribed spacers (ITS). G. graminis var. tritici, G. graminis var. avenae, and G. graminis var. graminis isolates can be distinguished from each other by nucleotide sequence differences in the ITS regions. The G. graminis var. tritici isolates can be further subdivided into R and N isolates (correlating with ability [R] or inability [N] to infect rye). Phylogenetic analysis of the ITS regions of several oat-infecting G. graminis var. tritici isolates suggests that these isolates are actually more closely related to G. graminis var. avenae. The isolates of Magnaporthe grisea included in the analysis showed a surprising degree of relatedness to members of the Gaeumannomyces-Phialophora complex. G. graminis variety-specific oligonucleotide primers were used in PCRs to amplify DNA from cereal seedlings infected with G. graminis var. tritici or G. graminis var. avenae, and these should be valuable for sensitive detection of pathogenic isolates and for diagnosis of take-all.

  7. Take-all or nothing.

    Science.gov (United States)

    Hernández-Restrepo, M; Groenewald, J Z; Elliott, M L; Canning, G; McMillan, V E; Crous, P W

    2016-01-01

    Take-all disease of Poaceae is caused by Gaeumannomyces graminis (Magnaporthaceae). Four varieties are recognised in G. graminis based on ascospore size, hyphopodial morphology and host preference. The aim of the present study was to clarify boundaries among species and varieties in Gaeumannomyces by combining morphology and multi-locus phylogenetic analyses based on partial gene sequences of ITS, LSU, tef1 and rpb1. Two new genera, Falciphoriella and Gaeumannomycella were subsequently introduced in Magnaporthaceae. The resulting phylogeny revealed several cryptic species previously overlooked within Gaeumannomyces. Isolates of Gaeumannomyces were distributed in four main clades, from which 19 species could be delimited, 12 of which were new to science. Our results show that the former varieties Gaeumannomyces graminis var. avenae and Gaeumannomyces graminis var. tritici represent species phylogenetically distinct from G. graminis, for which the new combinations G. avenae and G. tritici are introduced. Based on molecular data, morphology and host preferences, Gaeumannomyces graminis var. maydis is proposed as a synonym of G. radicicola. Furthermore, an epitype for Gaeumannomyces graminis var. avenae was designated to help stabilise the application of that name.

  8. Evaluation of a crataegus-based multiherb formula for dyslipidemia: a randomized, double-blind, placebo-controlled clinical trial.

    Science.gov (United States)

    Hu, Miao; Zeng, Weiwei; Tomlinson, Brian

    2014-01-01

    Background. We for the first time examined the effects of a multiherb formula containing Crataegus pinnatifida (1 g daily), Alisma orientalis, Stigma maydis, Ganoderma lucidum, Polygonum multiflorum, and Morus alba on plasma lipid and glucose levels in Chinese patients with dyslipidemia. Methods. In this randomized, double-blind, placebo-controlled study, 42 patients were randomized at a ratio of 1 : 1 to receive the herbal formula or placebo for 12 weeks and 40 patients completed the study. Lipid profiles, glucose, glycated haemoglobin (HbA1c), and laboratory safety parameters were performed before and after treatment. Results. The difference in the changes in low-density lipoprotein cholesterol (LDL-C) levels between placebo and active treatment (-9%) was significantly (P < 0.05) better with active treatment. HbA1c levels significantly decreased by -3.9% in the active treatment group, but the change was not significantly different from that with placebo (-1.1%) (P = 0.098). There were no apparent adverse effects or changes in laboratory safety parameters with either treatment. Conclusions. The multiherb formula had mild beneficial effects on plasma LDL-C after 12-weeks treatment in subjects with dyslipidemia without any noticeable adverse effects.

  9. Evaluation of a Crataegus-Based Multiherb Formula for Dyslipidemia: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial

    Directory of Open Access Journals (Sweden)

    Miao Hu

    2014-01-01

    Full Text Available Background. We for the first time examined the effects of a multiherb formula containing Crataegus pinnatifida (1 g daily, Alisma orientalis, Stigma maydis, Ganoderma lucidum, Polygonum multiflorum, and Morus alba on plasma lipid and glucose levels in Chinese patients with dyslipidemia. Methods. In this randomized, double-blind, placebo-controlled study, 42 patients were randomized at a ratio of 1 : 1 to receive the herbal formula or placebo for 12 weeks and 40 patients completed the study. Lipid profiles, glucose, glycated haemoglobin (HbA1c, and laboratory safety parameters were performed before and after treatment. Results. The difference in the changes in low-density lipoprotein cholesterol (LDL-C levels between placebo and active treatment (−9% was significantly (P<0.05 better with active treatment. HbA1c levels significantly decreased by −3.9% in the active treatment group, but the change was not significantly different from that with placebo (−1.1% (P=0.098. There were no apparent adverse effects or changes in laboratory safety parameters with either treatment. Conclusions. The multiherb formula had mild beneficial effects on plasma LDL-C after 12-weeks treatment in subjects with dyslipidemia without any noticeable adverse effects.

  10. [Genetic analysis of maize cytoplasmic male sterile mutants obtained by space flight].

    Science.gov (United States)

    Zhang, Cai-Bo; Yuan, Guo-Zhao; Wang, Jing; Pan, Guang-Tang; Rong, Ting-Zhao; Cao, Mo-Ju

    2011-02-01

    Three maize male sterile mutants were obtained from the offsprings of two maize inbred lines 18-599 and 08-641, which were carried into space by the Shijian 8 Satellite. The stability of male sterile expression was observed in different locations, years, and seasons. In order to analyze the genetic characteristic of male sterility, testcross, backcross and reciprocal cross were made with these male sterile plants. The results showed that the male sterility character was stable in different locations, years, and seasons, and the sterility was inheritable. Because the maintainer lines and restorer lines for these sterile materials were found, and there was no male sterile plant separated among the reciprocal cross F2. Thus, we concluded that these mutants could be cytoplasmic male sterile. Combining the results of male fertility restoration test and PCR analysis, we could conclude that the three male sterile mutants were classified into the CMS-C type in maize. Owing to their difference in fertility restoration, these mutants may belong to different subgroups of CMS-C type. The discovery of the three male sterile mutants increased the genetic diversity of CMS-C type, improved the tolerance to Bipolaris maydis, and laid a foundation for extensive application of CMS-C in seeds production.

  11. An antifungal peptide from Phaseolus vulgaris cv. brown kidney bean

    Institute of Scientific and Technical Information of China (English)

    Yau Sang Chan; Jack Ho Wong; Evandro Fei Fang; Wen Liang Pan; Tzi Bun Ng

    2012-01-01

    A 5.4-kDa antifungal peptide,with an N-terminal sequence highly homologous to defensins and inhibitory activity against Mycosphaerella arachidicola (IC5o=3 μM),Setospaeria turcica and Bipolaris maydis,was isolated from the seeds of Phaseolus vulgaris cv.brown kidney bean.The peptide was purified by employing a protocol that entailed adsorption on Affi-gel blue gel and Mono S and finally gel filtration on Superdex 75.The antifungal activity of the peptide against M.arachidicola was stable in the pH range 3-12 and in the temperature range 0℃ to 80℃.There was a slight reduction of the antifungal activity at pH 2 and 13,and the activity was indiscernible at pH 0,1,and 14.The activity at 90℃ and 100℃ was slightly diminished.Deposition of Congo red at the hyphal tips of M.arachidicola was induced by the peptide indicating inhibition of hyphal growth.The lack of antiproliferative activity of brown kidney bean antifungal peptide toward tumor cells,in contrast to the presence of such activity of other antifungal peptides,indicates that different domains are responsible for the antifungal and antiproliferative activities.

  12. Isolation and characterization of juncin, an antifungal protein from seeds of Japanese Takana (Brassica juncea Var. integrifolia).

    Science.gov (United States)

    Ye, Xiujuan; Ng, Tzi Bun

    2009-05-27

    An 18.9 kDa antifungal protein designated juncin was isolated from seeds of the Japanese takana (Brassica juncea var. integrifolia). The purification protocol employed comprised anion-exchange chromatography on Q-Sepharose, affinity chromatography on Affi-gel blue gel, cation exchange chromatography on SP-Sepharose, and gel filtration on Superdex 75. Juncin was adsorbed on Affi-gel blue gel and SP-Sepharose but unadsorbed on Q-Sepharose. The protein exhibited antifungal activity toward the phytopathogens Fusarium oxysporum, Helminthosporium maydis, and Mycosphaerella arachidicola with IC(50) values of 13.5, 27, and 10 μM, respectively. It was devoid of mitogenic activity toward splenocytes and nitric oxide inducing activity toward macrophages. It inhibited the proliferation of hepatoma (HepG2) and breast cancer (MCF7) cells with IC(50) values of 5.6 and 6.4 μM, respecitvely, and the activity of HIV-1 reverse transcriptase with an IC(50) of 4.5 μM. Its N-terminal sequence differed from those of antifungal proteins that have been reported to date. Compared with Brassica campestris and Brassica alboglabra antifungal peptides, juncin exhibits a different molecular mass and N-terminal amino acid sequence but similar biological activities.

  13. An antifungal peptide from Phaseolus vulgaris cv. brown kidney bean.

    Science.gov (United States)

    Chan, Yau Sang; Wong, Jack Ho; Fang, Evandro Fei; Pan, Wen Liang; Ng, Tzi Bun

    2012-04-01

    A 5.4-kDa antifungal peptide, with an N-terminal sequence highly homologous to defensins and inhibitory activity against Mycosphaerella arachidicola (IC(50)= 3 μM), Setospaeria turcica and Bipolaris maydis, was isolated from the seeds of Phaseolus vulgaris cv. brown kidney bean. The peptide was purified by employing a protocol that entailed adsorption on Affi-gel blue gel and Mono S and finally gel filtration on Superdex 75. The antifungal activity of the peptide against M. arachidicola was stable in the pH range 3-12 and in the temperature range 0°C to 80°C. There was a slight reduction of the antifungal activity at pH 2 and 13, and the activity was indiscernible at pH 0, 1, and 14. The activity at 90°C and 100°C was slightly diminished. Deposition of Congo red at the hyphal tips of M. arachidicola was induced by the peptide indicating inhibition of hyphal growth. The lack of antiproliferative activity of brown kidney bean antifungal peptide toward tumor cells, in contrast to the presence of such activity of other antifungal peptides, indicates that different domains are responsible for the antifungal and antiproliferative activities.

  14. Potencial de híbridos simples de milho para extração de linhagens Potential of maize single hybrids to generate inbred lines

    Directory of Open Access Journals (Sweden)

    Odair Bison

    2003-04-01

    Full Text Available A utilização de híbridos simples comerciais de milho é uma das opções de populações para a extração de linhagens, porque são adaptados e provavelmente concentram alta freqüência de alelos favoráveis já fixados. Mesmo nos locos que estão segregando, a freqüência de alelos favoráveis é 0,5. Assim, a identificação de populações promissoras, derivadas de híbridos simples superiores, é uma boa estratégia para aumentar a eficiência dos programas de melhoramento. As populações derivadas dos híbridos simples comerciais AG9012 e C333 foram avaliadas com o objetivo de verificar o potencial dessas para extração de linhagens superiores, por meio das estimativas de parâmetros genéticos e fenotípicos, da estimativa de m+a e a metodologia proposta por Jinks & Pooni (1976. Foram avaliadas 169 famílias S1 de cada população, durante a safra agrícola de 1999/2000, na área experimental do Departamento de Biologia da UFLA, em Lavras - MG, em látice simples 13x13, sendo as parcelas constituídas por uma linha de 3 m. As características analisadas foram incidência de Phaeosphaeria maydis em duas épocas, altura de plantas, altura de espigas e produtividade de espigas despalhadas. Foi constatado que há possibilidade de se obterem linhagens com bom desempenho per se, sendo a população derivada do C333 a mais promissora, por associar resistência a Phaeosphaeria maydis e possuir média mais alta e maior probabilidade de obtenção de linhagens superiores. A metodologia proposta por Jinks & Pooni (1976 mostrou-se mais informativa do que a estimativa de m+a para a escolha de populações, mas, quando possível, as duas podem ser utilizadas simultaneamente para auxiliar na decisão dos melhoristas.Populations derived from commercial single hybrids are one of the breeder options for inbred line extraction because of their adaptation and probable high frequency of loci with fixed favorable alleles. Even the segregating loci carry

  15. Diallel analyze of yield and progress of the severity of leaf diseases in maize hybrids in two population density

    Directory of Open Access Journals (Sweden)

    Marcos Ventura Faria

    2015-02-01

    Full Text Available Seven commercial maize hybrids (AS1575, 2B688, Penta, GNZ2004, AG8021, Sprint e P30F53 were intercrossed in a complete diallel, excluded reciprocal, obtaining 21 crosses. The 28 treatments were evaluated in two environments characterized by different densities (62,500 and 90,000 plants ha-1, with the aim of selecting the most promising parents for generating base population to obtain lines. Two experiments were carried out in Guarapuava-PR, at randomized block design with three replications. We estimated the general (GCA and specific (SCA combining abilities for yield and disease severity assessed by the area under the common rust (Puccinia sorghi progress curve (AURPC and the area under the leaf spot (Cercospora zeae-maydis progress curve (AULPC. The effects of GCA and SCA were significant for grain yield and diseases severity in both densities, revealing the importance of both additive and non-additive effects. There GCA x densities interaction was significant only for grain yield. Crossings P30F53 x AG8021 and P30F53 x Penta had negative estimates of SCA for AURPC and AULPC on the environments average. Hybrids GNZ 2004 and P30F53 stood out showing positive GCA for grain yield and negative for AURPC and AULPC in both densities and therefore are recommended for generating base populations for obtaining lines adapted for both densities, conventional and denser plantings, given the current trends in management of maize.

  16. Ketahanan Beberapa Jamur Patogen terhadap Fungisida

    Directory of Open Access Journals (Sweden)

    Christanti Sumardiyono

    1995-12-01

    Full Text Available Chemical control of plant pathogens have been done for a long time using contact and systemic fungicides. Resistance of the pathogens to fungicides may caused failure of disease control program. Studies by in vitro, in green house and fields were done at Yogyakarta to know the fungal which were resistant to several fungicides after treatment. The pathogen tested to fungicides were Colletotrichum capsici on red pepper to propineb, mancozeb, mixture of carbendazim and mancozeb, and thiophanate methyl, Alternaria porri on garlic to thiophanate methyl, Phytophthora palmivora on cocoa to Al-fosetyl, metalaxyl, mancozeb, and Copper Oxychloride, and Peronosclerospora maydis on corn to metalaxyl. The study indicated that C. capsici was resistant to propineb, mancozeb, and mixture of carbendazim and mancozeb, but sensitive to thiophanate methyl. A. porri from plant treated with thiophanate methyl indicated resistant to the fungicides. ED50 of Al-fosetyl and metalaxyl on P. palmivora were higher than on mancozeb and copper oxychloride, so that it was resistance strain. Key words: contact fungicides, systemic fungicides, resistance

  17. Identification of a Chitinase-modifying Protein from Fusarium verticillioides

    Science.gov (United States)

    Naumann, Todd A.; Wicklow, Donald T.; Price, Neil P. J.

    2011-01-01

    Chitinase-modifying proteins (cmps) are proteases secreted by fungal pathogens that truncate the plant class IV chitinases ChitA and ChitB during maize ear rot. cmp activity has been characterized for Bipolaris zeicola and Stenocarpella maydis, but the identities of the proteases are not known. Here, we report that cmps are secreted by multiple species from the genus Fusarium, that cmp from Fusarium verticillioides (Fv-cmp) is a fungalysin metalloprotease, and that it cleaves within a sequence that is conserved in class IV chitinases. Protein extracts from Fusarium cultures were found to truncate ChitA and ChitB in vitro. Based on this activity, Fv-cmp was purified from F. verticillioides. N-terminal sequencing of truncated ChitA and MALDI-TOF-MS analysis of reaction products showed that Fv-cmp is an endoprotease that cleaves a peptide bond on the C-terminal side of the lectin domain. The N-terminal sequence of purified Fv-cmp was determined and compared with a set of predicted proteins, resulting in its identification as a zinc metalloprotease of the fungalysin family. Recombinant Fv-cmp also truncated ChitA, confirming its identity, but had reduced activity, suggesting that the recombinant protease did not mature efficiently from its propeptide-containing precursor. This is the first report of a fungalysin that targets a nonstructural host protein and the first to implicate this class of virulence-related proteases in plant disease. PMID:21878653

  18. Identification of a chitinase-modifying protein from Fusarium verticillioides: truncation of a host resistance protein by a fungalysin metalloprotease.

    Science.gov (United States)

    Naumann, Todd A; Wicklow, Donald T; Price, Neil P J

    2011-10-14

    Chitinase-modifying proteins (cmps) are proteases secreted by fungal pathogens that truncate the plant class IV chitinases ChitA and ChitB during maize ear rot. cmp activity has been characterized for Bipolaris zeicola and Stenocarpella maydis, but the identities of the proteases are not known. Here, we report that cmps are secreted by multiple species from the genus Fusarium, that cmp from Fusarium verticillioides (Fv-cmp) is a fungalysin metalloprotease, and that it cleaves within a sequence that is conserved in class IV chitinases. Protein extracts from Fusarium cultures were found to truncate ChitA and ChitB in vitro. Based on this activity, Fv-cmp was purified from F. verticillioides. N-terminal sequencing of truncated ChitA and MALDI-TOF-MS analysis of reaction products showed that Fv-cmp is an endoprotease that cleaves a peptide bond on the C-terminal side of the lectin domain. The N-terminal sequence of purified Fv-cmp was determined and compared with a set of predicted proteins, resulting in its identification as a zinc metalloprotease of the fungalysin family. Recombinant Fv-cmp also truncated ChitA, confirming its identity, but had reduced activity, suggesting that the recombinant protease did not mature efficiently from its propeptide-containing precursor. This is the first report of a fungalysin that targets a nonstructural host protein and the first to implicate this class of virulence-related proteases in plant disease.

  19. Permanent Genetic Resources added to Molecular Ecology Resources database 1 January 2009-30 April 2009.

    Science.gov (United States)

    Abercrombie, L G; Anderson, C M; Baldwin, B G; Bang, I C; Beldade, R; Bernardi, G; Boubou, A; Branca, A; Bretagnolle, F; Bruford, M W; Buonamici, A; Burnett, R K; Canal, D; Cárdenas, H; Caullet, C; Chen, S Y; Chun, Y J; Cossu, C; Crane, C F; Cros-Arteil, S; Cudney-Bueno, R; Danti, R; Dávila, J A; Della Rocca, G; Dobata, S; Dunkle, L D; Dupas, S; Faure, N; Ferrero, M E; Fumanal, B; Gigot, G; González, I; Goodwin, S B; Groth, D; Hardesty, B D; Hasegawa, E; Hoffman, E A; Hou, M L; Jamsari, A F J; Ji, H J; Johnson, D H; Joseph, L; Justy, F; Kang, E J; Kaufmann, B; Kim, K S; Kim, W J; Koehler, A V; Laitung, B; Latch, P; Liu, Y D; Manjerovic, M B; Martel, E; Metcalfe, S S; Miller, J N; Midgley, J J; Migeon, A; Moore, A J; Moore, W L; Morris, V R F; Navajas, M; Navia, D; Neel, M C; De Nova, P J G; Olivieri, I; Omura, T; Othman, A S; Oudot-Canaff, J; Panthee, D R; Parkinson, C L; Patimah, I; Pérez-Galindo, C A; Pettengill, J B; Pfautsch, S; Piola, F; Potti, J; Poulin, R; Raimondi, P T; Rinehart, T A; Ruzainah, A; Sarver, S K; Scheffler, B E; Schneider, A R R; Silvain, J F; Siti Azizah, M N; Springer, Y P; Stewart, C N; Sun, W; Tiedemann, R; Tsuji, K; Trigiano, R N; Vendramin, G G; Wadl, P A; Wang, L; Wang, X; Watanabe, K; Waterman, J M; Weisser, W W; Westcott, D A; Wiesner, K R; Xu, X F; Yaegashi, S; Yuan, J S

    2009-09-01

    This article documents the addition of 283 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Agalinis acuta; Ambrosia artemisiifolia; Berula erecta; Casuarius casuarius; Cercospora zeae-maydis; Chorthippus parallelus; Conyza canadensis; Cotesia sesamiae; Epinephelus acanthistius; Ficedula hypoleuca; Grindelia hirsutula; Guadua angustifolia; Leucadendron rubrum; Maritrema novaezealandensis; Meretrix meretrix; Nilaparvata lugens; Oxyeleotris marmoratus; Phoxinus neogaeus; Pristomyrmex punctatus; Pseudobagrus brevicorpus; Seiridium cardinale; Stenopsyche marmorata; Tetranychus evansi and Xerus inauris. These loci were cross-tested on the following species: Agalinis decemloba; Agalinis tenella; Agalinis obtusifolia; Agalinis setacea; Agalinis skinneriana; Cercospora zeina; Cercospora kikuchii; Cercospora sorghi; Mycosphaerella graminicola; Setosphaeria turcica; Magnaporthe oryzae; Cotesia flavipes; Cotesia marginiventris; Grindelia Xpaludosa; Grindelia chiloensis; Grindelia fastigiata; Grindelia lanceolata; Grindelia squarrosa; Leucadendron coniferum; Leucadendron salicifolium; Leucadendron tinctum; Leucadendron meridianum; Laodelphax striatellus; Sogatella furcifera; Phoxinus eos; Phoxinus rigidus; Phoxinus brevispinosus; Phoxinus bicolor; Tetranychus urticae; Tetranychus turkestani; Tetranychus ludeni; Tetranychus neocaledonicus; Tetranychus amicus; Amphitetranychus viennensis; Eotetranychus rubiphilus; Eotetranychus tiliarium; Oligonychus perseae; Panonychus citri; Bryobia rubrioculus; Schizonobia bundi; Petrobia harti; Xerus princeps; Spermophilus tridecemlineatus and Sciurus carolinensis.

  20. [An orientational examination of the effects of extracts from mixtures of herbal drugs on selected renal functions].

    Science.gov (United States)

    Masteiková, R; Klimas, R; Samura, B B; Savickas, A; Samura, B A; Belaij, S I; Samura, I B; Rabisková, M; Chalupová, Z; Bernatoniene, J

    2007-04-01

    The paper aimed to determine the effects of mixtures of selected medicinal plants on some physiological renal functions, i.e. excretion of urine and electrolytes and changes in the quantity of prostaglandins E2 (PGE2) and kallikrein-kinins in rat blood plasma after water and salt load. The following medicinal plants were selected for the examination: downy birch (Betula pubescens EHRH.), everlasting flower (Helichrysum arenarium L. MOENCH.), hawthorn (Crataegus oxyacantha L.), woodland strawberry (Fragaria vesca L.), sweet corn (Zea mays L.), German chamomile (Matricaria recutita L.), and field horsetail (Equisetum arvense L.). Herbal drugs were used to compose 6 mixtures. Extracts from these mixtures were administered to Wistar strain males and their effects were compared with the effects of an administered suspension of hydrochlorothiazide, an extract from field horsetail herb alone, and a control group of animals which was not administered any preparation. The greatest diuretic effect was found in a mixture composed of birch leaves (Betulae folium), hawthorn berries (Crataegi fructus), strawberry leaves (Fragariae folium), corn silk (Maydis stigmata), chamomile flowers (Matricariae flos), and horsetail herb (Equiseti herba). Its effect was greater by 47% and 34% than the effect of a horsetail herb extract and a hydrochlorothiazide suspension (p < 0.05), respectively. The extract from this mixture also increased the quantity of prostaglandins E2 and kallikrein-kinins in rat blood plasma in water and salt load.

  1. Antifungal metabolites (monorden, monocillins I, II, III) from Colletotrichum graminicola, a systemic vascular pathogen of maize.

    Science.gov (United States)

    Wicklow, Donald T; Jordan, Annalisa M; Gloer, James B

    2009-12-01

    Colletotrichum graminicola is a systemic vascular pathogen that causes anthracnose stalk rot and leaf blight of maize. In the course of an effort to explore the potential presence and roles of C. graminicola metabolites in maize, ethyl acetate extracts of solid substrate fermentations of several C. graminicola isolates from Michigan and Illinois were found to be active against Aspergillus flavus and Fusarium verticillioides, both mycotoxin-producing seed-infecting fungal pathogens. Chemical investigations of the extract of one such isolate (NRRL 47511) led to the isolation of known metabolites monorden (also known as radicicol) and monocillins I-III as major components. Monorden and monocillin I displayed in vitro activity against the stalk- and ear-rot pathogen Stenocarpella maydis while only the most abundant metabolite (monorden) showed activity against foliar pathogens Alternaria alternata, Bipolaris zeicola, and Curvularia lunata. Using LC-HRESITOFMS, monorden was detected in steam-sterilized maize stalks and stalk residues inoculated with C. graminicola but not in the necrotic stalk tissues of wound-inoculated plants grown in an environmental chamber. Monorden and monocillin I can bind and inhibit plant Hsp90, a chaperone of R-proteins. It is hypothesized that monorden and monocillins could support the C. graminicola disease cycle by disrupting maize plant defenses and by excluding other fungi from necrotic tissues and crop residues. This is the first report of natural products from C. graminicola, as well as the production of monorden and monocillins by a pathogen of cereals.

  2. IDENTIFICATION OF SPECIES BF-02 AND ITS ANTIFUNGAL ACTIVITY AGAINST SEVERAL PLANT PATHOGENS%拮抗菌Bf-02菌株鉴定及对几种植物病原真菌抑制活性测定

    Institute of Scientific and Technical Information of China (English)

    王清海; 牛赡光; 刘幸红; 刘玉升

    2010-01-01

    在对土壤有益微生物的分离、筛选中获得一株拮抗细菌Bf-02,该菌对多种植物病原真菌具有较强的抑菌活性.通过对菌株的培养性状、形态特征、生理生化特性等试验项目,初步鉴定该菌株为坚强芽孢杆菌(Bacillus firmus).采用平板对峙培养法测定Bf-02菌株抑菌活性,结果表明Bf-02对供试的14种植物病原真菌,除花生白绢病菌(Sclerotium rolfsii)外,都具有抑菌活性.其中对核桃炭疽病菌(Colletotrichum gloeosporioides)、小麦根腐病菌(Bipolaris sorokiniana)、玉米小斑病菌(Bipolaris maydis)效果最好,与其他处理在P<0.05水平上具有显著性差异.

  3. Conservation of XYN11A and XYN11B xylanase genes in Bipolaris sorghicola, Cochliobolus sativus, Cochliobolus heterostrophus, and Cochliobolus spicifer.

    Science.gov (United States)

    Emami, Kaveh; Hack, Ethan

    2002-10-01

    Two types of xylanase gene, XYN11A ( XYL1) and XYN11B ( XYL2), were amplified by PCR and partially sequenced in four phytopathogenic species of the ascomycete fungal genus Cochliobolus (anamorph genus Bipolaris). Three of the species, C. heterostrophus ( B. maydis), C. sativus ( B. sorokiniana), and Bipolaris sorghicola (no teleomorph known), are interrelated; the fourth, C. spicifer ( B. spicifera), was found, through analysis of the 5.8S RNA and internal transcribed spacer (ITS) sequences of its ribosomal DNA, to be more distantly related to the other three. Isolates from all four species contain orthologous XYN11A and XYN11B genes, but a set of laboratory strains of C. heterostrophus gave no product corresponding to the XYN11B gene. The patterns of evolution of the two xylanase genes and ribosomal DNA sequences are mutually consistent; the results indicate that the two genes were present in the common ancestor of all Cochliobolus species and are evolving independently of each other.

  4. Cytogenetic Mapping of Disease Resistance Genes and Analysis of Their Distribution Features on Chromosomes in Maize

    Institute of Scientific and Technical Information of China (English)

    LiLi-jia; SongYun-chun

    2003-01-01

    Cytogenetic maps of four clusters of disease resistance genes were generated by ISH of the two RFLP markers tightly linked to and flanking each of maize resistance genes and the cloned resistance genes from other plant species onto maize chromosomes, combining with data published before. These genes include Helminthosporium turcium Pass resistance genes Htl, Htnl and Ht2, Helminthosporium maydis Nisik resistance genes Rhml and Rhm2,maize dwarf mosaic virus resistance gene Mdml, wheat streak mosaic virus resistance gene Wsml, Helminthosporium carbonum ULLstrup resistance gene Hml and the cloned Xanthomonas oryzae pv. Oryzae resistance gene Xa21 of rice, Cladosporium fulvum resistance genes Cf-9 and Cf-2. 1 of tomato, and Pseudomonas syringae resistance gene RPS2 of Arabidopsis. Most of the tested disease resistance genes located on the four chromosomes, i. e. , chromosomesl, 3, 6 and 8, and they closely distributed at the interstitial regions of these chromosomal long arms with percentage distances ranging 31.44(±3.72)-72.40(±3. 25) except for genes Rhml, Rhm2, Mdml and Wsml which mapped on the satellites of the short arms of chromosome6. It showed that the tested RFLP markers and genes were duplicated or triplicated in maize genome. Homology and conservation of disease resistance genes among species, and relationship between distribution features and functions of the genes were discussed. The results provide important scientific basis for deeply understanding structure and function of disease resistance genes and breeding in maize.

  5. APPLICATION OF FOLIAR FERTILIZER AND FUNGICIDES ON WHITE SPOT DISEASE CONTROL AND DEVELOPMENT OF MAIZE

    Directory of Open Access Journals (Sweden)

    C. H. Brito

    2016-11-01

    Full Text Available This study aimed to evaluate the effect of forms of application of foliar fertilizers and fungicides to control fungus causing white maize spot, Phaeosphaeria maydis, and the growth and development of hybrids maize. The design had randomized blocks, with the use two sources hybrids maize with different reaction to white maize spot: resistant and susceptive and application with moisture of foliar chemical fungicide + cobalt foliar + molybdenum foliar + manganese and control treatment, without application consisting of four treatments and five replications. In the treatments with application of moisture were done on vegetative maize growth stage V8 (stage that determine that the number of kernel rows, VT (stage that arrives when the last branch of the tassel is completely visible and reproductive maize growth stage R2 (kernels are white on the outside and resemble a blister. The evaluation of variables of growth of maize plants: grain dry mass, cob dry mass, leaf dry mass, culm dry mass and modificated leaf ear and total plant dry mass. Harvest was carried out when the grains were 20% humidity. The application of fungicides and foliar fertilizers increased the leaves, culm, ear modificated, cobs and shoot dry mass plants maize. The application of fungicides and foliar fertilizers providing returning of 11.409,5 kg ha-1 of shoot dry mass plants with increased of 1.296 kg ha-1 on soil(12,81%. In the susceptive hybrid maize the application of fungicides and foliar fertilizers provide higher dry mass grains and shoot of plants.

  6. Evaluation of a Crataegus-Based Multiherb Formula for Dyslipidemia: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial

    Science.gov (United States)

    Zeng, Weiwei; Tomlinson, Brian

    2014-01-01

    Background. We for the first time examined the effects of a multiherb formula containing Crataegus pinnatifida (1 g daily), Alisma orientalis, Stigma maydis, Ganoderma lucidum, Polygonum multiflorum, and Morus alba on plasma lipid and glucose levels in Chinese patients with dyslipidemia. Methods. In this randomized, double-blind, placebo-controlled study, 42 patients were randomized at a ratio of 1 : 1 to receive the herbal formula or placebo for 12 weeks and 40 patients completed the study. Lipid profiles, glucose, glycated haemoglobin (HbA1c), and laboratory safety parameters were performed before and after treatment. Results. The difference in the changes in low-density lipoprotein cholesterol (LDL-C) levels between placebo and active treatment (−9%) was significantly (P < 0.05) better with active treatment. HbA1c levels significantly decreased by −3.9% in the active treatment group, but the change was not significantly different from that with placebo (−1.1%) (P = 0.098). There were no apparent adverse effects or changes in laboratory safety parameters with either treatment. Conclusions. The multiherb formula had mild beneficial effects on plasma LDL-C after 12-weeks treatment in subjects with dyslipidemia without any noticeable adverse effects. PMID:24834096

  7. Genetic characterization and linkage disequilibrium mapping of resistance to gray leaf spot in maize(Zea mays L.)

    Institute of Scientific and Technical Information of China (English)

    Liyu; Shi; Xiangling; Lv; Jianfeng; Weng; Hanyong; Zhu; Changlin; Liu; Zhuanfang; Hao; Yu; Zhou; Degui; Zhang; Mingshun; Li; Xiaoke; Ci; Xinhai; Li; Shihuang; Zhang

    2014-01-01

    Gray leaf spot(GLS),caused by Cercospora zeae-maydis,is an important foliar disease of maize(Zea mays L.)worldwide,resistance to which is controlled by multiple quantitative trait loci(QTL).To gain insights into the genetic architecture underlying the resistance to this disease,an association mapping population consisting of 161 inbred lines was evaluated for resistance to GLS in a plant pathology nursery at Shenyang in 2010 and 2011.Subsequently,a genome-wide association study,using 41,101 single-nucleotide polymorphisms(SNPs),identified 51 SNPs significantly(P<0.001)associated with GLS resistance,which could be converted into 31 QTL.In addition,three candidate genes related to plant defense were identified,including nucleotidebinding-site/leucine-rich repeat,receptor-like kinase genes similar to those involved in basal defense.Two genic SNPs,PZE-103142893 and PZE-109119001,associated with GLS resistance in chromosome bins 3.07 and 9.07,can be used for marker-assisted selection(MAS)of GLS resistance.These results provide an important resource for developing molecular markers closely linked with the target trait,enhancing breeding efficiency.

  8. Antimicrobial activity of pyrrocidines from Acremonium zeae against endophytes and pathogens of maize.

    Science.gov (United States)

    Wicklow, Donald T; Poling, Stephen M

    2009-01-01

    Acremonium zeae produces pyrrocidines A and B, which are polyketide-amino acid-derived antibiotics, and is recognized as a seedborne protective endophyte of maize which augments host defenses against microbial pathogens causing seedling blights and stalk rots. Pyrrocidine A displayed significant in vitro activity against Aspergillus flavus and Fusarium verticillioides in assays performed using conidia as inoculum, with pyrrocidine A being more active than B. In equivalent assays performed with conidia or hyphal cells as inoculum, pyrrocidine A revealed potent activity against major stalk and ear rot pathogens of maize, including F. graminearum, Nigrospora oryzae, Stenocarpella (Diplodia) maydis, and Rhizoctonia zeae. Pyrrocidine A displayed significant activity against seed-rotting saprophytes A. flavus and Eupenicillium ochrosalmoneum, as well as seed-infecting colonists of the phylloplane Alternaria alternata, Cladosporium cladosporioides, and Curvularia lunata, which produces a damaging leaf spot disease. Protective endophytes, including mycoparasites which grow asymptomatically within healthy maize tissues, show little sensitivity to pyrrocidines. Pyrrocidine A also exhibited potent activity against Clavibacter michiganense subsp. nebraskense, causal agent of Goss's bacterial wilt of maize, and Bacillus mojaviense and Pseudomonas fluorescens, maize endophytes applied as biocontrol agents, but were ineffective against the wilt-producing bacterium Pantoea stewartii.

  9. A novel and exploitable antifungal peptide from kale (Brassica alboglabra) seeds.

    Science.gov (United States)

    Lin, Peng; Ng, Tzi Bun

    2008-10-01

    The aim of this study was to purify and characterize antifungal peptides from kale seeds in view of the paucity of information on antifungal peptides from the family Brassicaceae, and to compare its characteristics with those of published Brassica antifungal peptides. A 5907-Da antifungal peptide was isolated from kale seeds. The isolation procedure comprised affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose and Mono S, and gel filtration on Superdex Peptide. The peptide was adsorbed on the first three chromatographic media. It inhibited mycelial growth in a number of fungal species including Fusarium oxysporum, Helminthosporium maydis, Mycosphaerella arachidicola and Valsa mali, with an IC(50) of 4.3microM, 2.1microM, 2.4microM, and 0.15microM, respectively and exhibited pronounced thermostability and pH stability. It inhibited proliferation of hepatoma (HepG2) and breast cancer (MCF7) cells with an IC(50) of 2.7microM and 3.4microM, and the activity of HIV-1 reverse transcriptase with an IC(50) of 4.9microM. Its N-terminal sequence differed from those of antifungal proteins which have been reported to date.

  10. 广西甘蔗主要真菌病害调查初报%Preliminary report on investigating major sugarcane fungal diseases in Guangxi

    Institute of Scientific and Technical Information of China (English)

    韦金菊; 邓展云; 黄诚华; 黄伟华; 颜梅新; 刘海斌; 唐红琴

    2012-01-01

    [Objective]The purpose of the present study was to determine the species and the occurrence status of major sugarcane fugal diseases in sugarcane plantation areas of Guangxi in order to provide scientific references for controlling sugarcane diseases. [Method]In 2009-2011, species and occurrence degrees of major sugarcane fugal diseases in sugarcane plantation areas of Guangxi were investigated through randomized surveys, fanner interviews, literature references, and location investigations; diseased sugarcane samples were collected and their species were identified through pathogen isolated culture and other relevant methods. [Result]Thirteen species of fungal diseases were identified, which were Thielaviopsis paradoxa (de Seynes) V. Hohnel, Ustilago scitaminea Sydow, Fusarium moniliforme Sheldon, Puccinia melanocephala H.&P. Syd., Stagonospora sacchari Lo et Ling, Colletotrichum falcatum Went, Phyllosticta saccharicola P. Henn., Pellicularia sasakii (Shirai) Ito, Elsinoe sacchari L., Helminthosporium sacchari (Breda de Haan) Butler, My-covellosiella koepkei (kruger) Deighton, Bipolaris stenospilum (Drechs.) Shoem., and Cercospora longipes Butler. Out of all of the fungal disease characteristics, smut, rust, tip rot, and pineapple disease were the most rampantly detrimental to sugarcane quality and yield. [ Conclusion ] Ustilago scitaminea Sydow turned out to be the most serious sugarcane disease; it is urgent to begin breeding highly-disease-resistant sugarcane varieties with elite field and environment adaptations to replace the current susceptible-to-diseases sugarcane varieties in order to increase production.%[目的]明确广西蔗区主要真菌病害的发生情况和种类,为甘蔗病害综合防治提供科学依据.[方法]2009~2011年采用随机踏查、种植户随访、资料查询和定点调查等方法对广西蔗区甘蔗真菌病害种类及发生程度进行调查,并采集病害样本,通过病原分离培养鉴定等方法确定真菌病

  11. Pseudozyma vetiver sp. nov., a novel anamorphic ustilaginomycetous yeast species isolated from the phylloplane in Thailand.

    Science.gov (United States)

    Chamnanpa, Thunnicha; Limtong, Pitayakon; Srisuk, Nantana; Limtong, Savitree

    2013-11-01

    Three strains representing one novel yeast species were isolated from the phylloplanes of the vetiver grasses (DMKU-LV90 and DMKU-LV99(T)) and sugarcane (DMKU-SP260) collected in Thailand by leaf washing followed by a plating technique. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and the sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer region (ITS), the three strains were found to represent a single novel anamorphic ustilaginomycetous yeast species in the genus Pseudozyma. The name Pseudozyma vetiver sp. nov. is proposed for this novel species. The type strain is DMKU-LV99(T) (BCC 61021 = CBS 12824). The novel species showed phylogenetic relationships to the other members of the genus Pseudozyma and to teleomorphic fungal genera, namely Ustilago, Sporisorium and Anomalomyces in Ustilaginaceae, Ustilaginales. The three strains showed identical sequences both in the D1/D2 and ITS regions. The Pseudozyma species closest to the novel species in terms of pairwise sequence similarity in the D1/D2 region was Pseudozyma pruni but with 2.3 % nucleotide substitutions (14 nucleotide substitutions and no gaps out of 606 nt). The novel species and P. pruni differed by 10.9 % nucleotide substitutions (75 nucleotide substitutions and 31 gaps out of 691 nt) in the ITS region. The phylogenetic analysis based on the combined sequences of the ITS region and the D1/D2 region of the LSU rRNA gene showed that the novel species was found to be most closely related to Pseudozyma fusiformata but with 2.9 % nucleotide substitutions in the D1/D2 region and 7.4 % nucleotide substitutions in the ITS region.

  12. Fungal spores in four catholic churches in the metropolitan area of Monterrey, Nuevo León State, Mexico – First study

    Directory of Open Access Journals (Sweden)

    Alejandra Rocha Estrada

    2015-05-01

    Full Text Available Introduction. About 500,000 species of fungi have been described to-date, although an estimated between 1 – 1.5 million species may occur. They have a wide distribution in nature, contributing to the decomposition of organic matter and playing a part in the biogeochemical cycles of major nutrients. A small number are considered pathogens of animals and plants. There is ample historical evidence that certain types of allergies are associated with fungi; exposure to fungal allergens occurs in both outdoor and indoor spaces. Many indoor allergens are the same as those found outside buildings, entering through windows and doors, ventilation systems, or through cracks or other fissures in the walls. Objective. To determine the diversity and abundance of fungal spores inside four churches in the metropolitan area of Monterrey city in Mexico. Materials and methods. The study was carried out from July 2009 – January 2010 using a Hirst type volumetric collector (Burkard Manufacturing Co Ltd. Results. A total of 31,629 spores from 54 taxa were registered in the four churches. The building that showed the highest amount of spores was the Santa Catarina Mártir Church with 12,766 spores, followed by Cristo Rey with 7,155 and Nuestra Señora del Roble with 6,887. Regularly high concentrations of spores were recorded from 14:00 – 20:00 hours. The highest concentration value was observed at the church of Santa Catarina Mártir at 16:00 hours with 1153 spores/m 3 air. Conclusions. The most abundant spores in the four churches studied corresponded to Cladosporium, the [i]Aspergillus/Penicillium complex[/i], [i]Coprinus[/i], [i]Ganoderma[/i], [i]Curvularia and Ustilago[/i].

  13. Assessment of fungal diversity in a water-damaged office building.

    Science.gov (United States)

    Green, Brett J; Lemons, Angela R; Park, Yeonmi; Cox-Ganser, Jean M; Park, Ju-Hyeong

    2017-04-01

    Recent studies have described fungal communities in indoor environments using gene sequencing-based approaches. In this study, dust-borne fungal communities were elucidated from a water-damaged office building located in the northeastern region of the United States using internal transcribed spacer (ITS) rRNA gene sequencing. Genomic DNA was extracted from 5 mg of floor dust derived from 22 samples collected from either the lower floors (n = 8) or a top floor (n = 14) of the office building. ITS gene sequencing resolved a total of 933 ITS sequences and was clustered into 216 fungal operational taxonomic units (OTUs). Analysis of fungal OTUs at the 97% similarity threshold showed a difference between the lower and top floors that was marginally significant (p = 0.049). Species richness and diversity indices were reduced in the lower floor samples compared to the top floor samples and there was a high degree of compositional dissimilarity within and between the two different areas within the building. Fungal OTUs were placed in the phyla Ascomycota (55%), Basidiomycota (41%), Zygomycota (3%), Glomeromycota (0.4%), Chytridiomycota (0.3%), and unassigned fungi (0.5%). The Ascomycota classes with the highest relative abundances included the Dothideomycetes (30%) and Eurotiomycetes (16%). The Basidiomycota consisted of the classes Ustilaginomycetes (14%), Tremellomycetes (11%), and Agaricomycetes (8%). Sequence reads derived from the plant pathogen Ustilago syntherismae were the most abundant in the analysis as were obligate Basidiomycota yeast species that accounted for 12% and 11% of fungal ITS sequences, respectively. ITS gene sequencing provides additional insight into the diversity of fungal OTUs. These data further highlight the contribution of fungi placed in the phylum Basidiomycota, obligate yeasts, as well as xerophilic species that are typically not resolved using traditional culture methods.

  14. Testing of Seedborne Fungi in Wheat Germplasm Conserved in the National Crop Genebank of China

    Institute of Scientific and Technical Information of China (English)

    DUAN Can-xing; WANG Xiao-ming; ZHU Zhen-dong; WU Xiao-fei

    2007-01-01

    There is a primary understanding of the dominant fungi in wheat seeds conserved in the National Crop Genebank of China (NCGB) and an evaluation of the healthy status of wheat germplasm propagated in different regions. A total of 1465 wheat accessions were detected for seedbone fungi by blotter, agar plate, and wash tests. By blotter test, 17 genera of fungi, including more than 30 species, were detected in 712 wheat accessions from Shaanxi, Hebei, and Qinghai provinces, China. Alternaria was the most frequently detected in wheat seeds from Shaanxi Province, followed by Rhizopus, Penicillium, Aspergillus, Bipolaris, Cladosporium, Gonatobotrys, Chaetomium, and others. Seedborne fungi in wheat seeds from Hebei Province, with relatively high incidence were Alternaria, Rhizopus, Penicillium, Aspergillus, Bipolaris, Cladosporium, and Fusarium. In the seeds from Qinghai Province, Alternaria, Rhizopus, Bipolaris, Cladosporium, and Trichothecium are important seed-borne fungi. The seed germination was reduced substantially when seeds were infected by Fusarium verticillioides (syn. F. moniliforme), Bipolaris nodulosa, and Cladosporium herbarum. Eighteen genera and 25 species of fungi were identified in 353 accessions from Shaanxi Province using the agar plate test. The dominant fungi were Alternaria, Aspergillus, Bipolaris, Gonatobotrys, Penicillium, and Fusarium. The smut fungi, Ustilago tritici, was detected by the wash test in 400 accessions, but it was low in incidence in 300 seed samples from Shaanxi Province (1.3%), and in 100 samples from Hebei Province (2.0%). Totally 19 genera of fungi were detected in wheat seed samples, and some seedborne fungi were saprophytic and others were biotrophic which could cause seedborne diseases in the field.

  15. TÉCNICAS PARA EL ESTUDIO DE LA INTERACCIÓN CAÑA DE AZÚCAR- Sporisorium scitamineum

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    Mar\\u00EDa La O-Hechavarr\\u00EDa

    2011-01-01

    Full Text Available El objetivo del presente trabajo fue revisar las herramientas y técnicas de laboratorio empleadas para conocer la interacción entre la caña de azúcar y Sporisorium scitamineum (Syd. M. Piepenbr., M. Stoll & F. Oberw. (Ustilago scitaminea Sydow, agente causal del carbón. La resistencia a esta enfermedad, en la caña, está dada por diversos factores, entre los que se destacan, la activación de numerosas proteínas de resistencia además de otros genes relacionados con el proceso de patogénesis. Para el estudio de esta interacción se han utilizado diferentes herramientas, entre las que se encuentran: las histológicas, en las que se evalúan los trastornos de la célula ante la penetración del hongo, las bioquímicas, las cuales se caracterizan por cambios en la composición de diferentes sustancias antimicrobianas; las histoquímicas a través del marcaje in situ de proteínas específicas y las moleculares que permiten la identificación de transcriptos que se activan o reprimen durante la interacción. Estas técnicas contribuyen a un mejor conocimiento de los mecanismos que participan durante la respuesta defensiva de la planta. Ello puede sentar las bases para el mejoramiento genético y además de poder ser utilizado como indicador en los programas de selección asistida por marcadores moleculares.

  16. Influence of key residues on the heterologous extracellular production of fungal ribonuclease U2 in the yeast Pichia pastoris.

    Science.gov (United States)

    Alvarez-García, Elisa; García-Ortega, Lucía; De los Ríos, Vivian; Gavilanes, José G; Martínez-del-Pozo, Alvaro

    2009-06-01

    Ribonuclease U2, secreted by the smut fungus Ustilago sphaerogena, is a cyclizing ribonuclease that displays a rather unusual specificity within the group of microbial extracellular RNases, best represented by RNase T1. Superposition of the three-dimensional structures of RNases T1 and U2 suggests that the RNase U2 His 101 would be the residue equivalent to the RNase T1 catalytically essential His 92. RNase U2 contains three disulfide bridges but only two of them are conserved among the family of fungal extracellular RNases. The non-conserved disulfide bond is established between Cys residues 1 and 54. Mispairing of the disulfide network due to the presence of two consecutive Cys residues (54 and 55) has been invoked to explain the presence of wrongly folded RNase U2 species when produced in Pichia pastoris. In order to study both hypotheses, the RNase U2 H101Q and C1/54S variants have been produced, purified, and characterized. The results obtained support the major conclusion that His 101 is required for proper protein folding when secreted by the yeast P. pastoris. On the other hand, substitution of the first Cys residue for Ser results in a mutant version which is more efficiently processed in terms of a more complete removal of the yeast alpha-factor signal peptide. In addition, it has been shown that elimination of the Cys 1-Cys 54 disulfide bridge does not interfere with RNase U2 proper folding, generating a natively folded but much less stable protein.

  17. Simple sequence repeat markers useful for sorghum downy mildew (Peronosclerospora sorghi and related species

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    Odvody Gary N

    2008-11-01

    Full Text Available Abstract Background A recent outbreak of sorghum downy mildew in Texas has led to the discovery of both metalaxyl resistance and a new pathotype in the causal organism, Peronosclerospora sorghi. These observations and the difficulty in resolving among phylogenetically related downy mildew pathogens dramatically point out the need for simply scored markers in order to differentiate among isolates and species, and to study the population structure within these obligate oomycetes. Here we present the initial results from the use of a biotin capture method to discover, clone and develop PCR primers that permit the use of simple sequence repeats (microsatellites to detect differences at the DNA level. Results Among the 55 primers pairs designed from clones from pathotype 3 of P. sorghi, 36 flanked microsatellite loci containing simple repeats, including 28 (55% with dinucleotide repeats and 6 (11% with trinucleotide repeats. A total of 22 microsatellites with CA/AC or GT/TG repeats were the most abundant (40% and GA/AG or CT/TC types contribute 15% in our collection. When used to amplify DNA from 19 isolates from P. sorghi, as well as from 5 related species that cause downy mildew on other hosts, the number of different bands detected for each SSR primer pair using a LI-COR- DNA Analyzer ranged from two to eight. Successful cross-amplification for 12 primer pairs studied in detail using DNA from downy mildews that attack maize (P. maydis & P. philippinensis, sugar cane (P. sacchari, pearl millet (Sclerospora graminicola and rose (Peronospora sparsa indicate that the flanking regions are conserved in all these species. A total of 15 SSR amplicons unique to P. philippinensis (one of the potential threats to US maize production were detected, and these have potential for development of diagnostic tests. A total of 260 alleles were obtained using 54 microsatellites primer combinations, with an average of 4.8 polymorphic markers per SSR across 34

  18. HETEROSE E CAPACIDADE COMBINATÓRIA ENTRE VARIEDADES DE MILHO EM SOLO ÁCIDO HETEROSIS AND COMBINING ABILITY AMONG VARIETIES OF MAIZE IN ACID SOIL

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    José Branco de Miranda Filho

    2007-09-01

    Full Text Available

    Cultivares de milho (Zea mays L. com tolerância à acidez e resistência a doenças têm viabilizado o cultivo em solos sob vegetação de cerrado. A identificação de germoplasma com potencial para o melhoramento, com essas características, foi o objetivo deste estudo. Dez variedades de milho, resistentes a Puccinia polysora, Phaeosphaeria maydis, Physopella zeae, Exserohilum turcicum e complexo enfezamento, foram avaliadas em cruzamentos  dialélicos. As variedades e seus híbridos foram avaliados em solo ácido e com baixa fertilidade. Os caracteres analisados foram: peso de espigas (PE, altura da planta (AP e altura da espiga (AE. As variedades PMI 8701, CMS 57NF, PMI 9401, CMS 58ND e AMARILLO DENTADO DMR, per se, produziram acima de 3,5 t/ha, o que equivale a mais de 70% em relação à testemunha. A heterose foi significativa para PE, AP e AE, e, entre seus componentes, também o foram a heterose específica para AE e a heterose média para PE. Os efeitos de capacidade geral de combinação (gi foram significativos para todos os caracteres, e suas maiores estimativas para PE (t/ha foram as de PMI 8701 (0,150, BR 105 (0,120 e CMS 59 (0,106. O híbrido interpopulacional CMS 57NF x PMI 8701, com produção média de 5,11 t/ha e 23,7% de heterose, pode ser indicado como um grupo heterótico potencial. As variedades PMI 8701, CMS 57NF, PMI 9401 e CMS 58ND foram consideradas promissoras para programas de melhoramento intrapopulacional nas condições de solo ácido.

    PLAVRAS-CHAVE: Heterose; estresse de acidez; capacidade de combinação; germoplasma.

    Maize (Zea mays L. cultivars tolerant to acidity and resistant to diseases have made the crop feasible in ";cerrado"; (savanna soil. The identification of potentially useful germplasm with the above mentioned characteristics

  19. Synthesis and screening for potential against phytopathogenic fungi activity of novel amides%新型酰胺类化合物的合成及抗植物病原真菌活性研究

    Institute of Scientific and Technical Information of China (English)

    周国萍; 刘伟; 金洪; 陶科; 侯太平

    2012-01-01

    In order to search for novel agrochemicals with potential anti-phythopathogenic fungi activity, a series of daphneone analogues were designed and synthesized. They were screened for antifungal activities against five phytopathogenic fungi; Rhizoctonia solani, Botrytis cirerea , Gibberella zeae , Bipolaris maydis, and Sclerotia sclerotium. The preliminary bioassays indicated that some compounds exhibited a fairly good activity. N-(2-fluorophenyl)-2, 4, 5-trimethyl-3-furancarboxamide (p) showed a strong fungistatic activity against R. solani (98% and 99% growth inhibition at 20 and 200 mg/L, respectively). Two compounds, N-(4-fluorophenyl)-2, 5-dimethyl-3-furancarboxamide (h) and N-(2-fluorophenyl)-2, 5-dimethyl-3-furancarboxamide (k) at 200 mg/L inhibited the growth of Sclerotia sclerotium at 94% and 90% , respectively. The EC50 values for compound k were 0. 034 mg/L, while for the control fungicide carbendazim were 0. 050 mg/L. According to the EC50 and the preventive activity of compound k, it can be inferred that compound k had a very good activity against Rhizoctonia solani. Thus, the compound k -was demonstrated to be the most promising candidate for further study.%为了寻找具有潜在抗真菌活性的新型农用化学品,我们以瑞香狼毒中提取的二苯酮类似物为先导,设计并合成了一系列的酰胺类化合物,并进行了以下5种植物真菌的筛选:水稻纹枯病菌(Rhizoctonia solani)、小麦赤霉病菌(Gibberella zeae)、玉米小斑病菌(Bipolaris maydis)、番茄灰霉病菌(Botrytis cirerea)和油菜菌核病菌(Sclerotinia sclerotiorum).初步的活性筛选研究显示:氮-(2-氟苯基)-2,4,5-三甲基-3-呋喃甲酰胺(p)具有很强的抗水稻纹枯病菌活性(在20和200 mg/L的浓度下抑制率分别为98%和99%);氮-(4-氟苯基)-2,5-二甲基-3-呋喃甲酰胺(h)和氮-(2-氟苯基)-2,5-二甲基-3-呋喃甲酰胺(k)这两种化合物在200 mg/L浓度下对油菜菌核病菌的抑制率分别为94%和90

  20. Cruzamentos dialélicos de linhagens de milho sob condições de mancha de Phaeosphaeria Diallel crosses among maize lines under Phaeosphaeria leaf spot infestation

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    Maria Elisa Ayres Guidetti Zagatto Paterniani

    2000-06-01

    Full Text Available Híbridos simples de milho resultantes de cruzamentos dialélicos entre dez linhagens endogâmicas provenientes do CIMMYT foram avaliados em 4 locais do Estado de São Paulo (Estações Experimentais do IAC, em 1996/97, quanto à estabilidade e adaptabilidade da produtividade de grãos. Os ensaios foram instalados sob delineamento de blocos casualizados com três repetições, incluindo quatro testemunhas comerciais. Foi verificada grande previsibilidade da produção e adaptabilidade ampla (b=1,0 da maioria dos híbridos, indicando que o conjunto de linhagens é fonte potencial de híbridos heteróticos e de estabilidade satisfatória. Houve intensa severidade da mancha foliar de Phaeosphaeria maydis em Ribeirão Preto, considerada um dos fatores de estresse que mais contribuiu para a redução da qualidade ambiental. A doença foi avaliada cerca de 30 dias após o florescimento, através de uma escala de notas de 1 a 9, correspondendo a 0% e >80% de aréa foliar afetada na planta adulta, respectivamente. Houve uma tendência dos materiais mais resistentes à doença apresentarem valores de adaptabilidade menores que 1,0 (bSingle-cross hybrids of maize obtained from diallell crosses among ten inbred lines from CIMMYT were grown in four locations of the State of São Paulo, Brazil, during 1996/97, to evaluate stability and adaptability of grain yield. The experiments were set up in a randomized block design with three replications, including four comercial controls. High predictability of grain yield and general adaptability (b=1,0 was observed for most hybrids, indicating that the lines are a potential source of heterosis as well as for good adaptability. A severe infestation of Phaeosphaeria maydis leaf spot occured in Ribeirão Preto, representing a stress factor responsible for reduction of environmental quality. The disease was assessed about 30 days after flowering, using a scale of grades of 1 to 9, for 0% and 80% of the leaf area

  1. Desempenho esempenho agronômico de novos híbridos de milho-pipoca no Noroeste do Estado do Paraná, Brasil = Agronomic performance of new popcorn hybrids in Northwestern Paraná state, Brazil

    Directory of Open Access Journals (Sweden)

    Rafael Augusto Vieira

    2009-01-01

    Full Text Available Em milho, variações de desempenho agronômico são comuns em ambientesdistintos, havendo resposta ainda mais diferenciada em híbridos simples. Este trabalho teve como objetivo avaliar o desempenho de novos híbridos simples de milho-pipoca, desenvolvidos pelo Programa de Melhoramento da Universidade Estadual de Maringá, no noroeste do Paraná. Para isso, avaliaram-se as características agronômicas de rendimento de grãos, capacidade de expansão, altura de planta, altura de espiga, resistência à ferrugem polissora (Puccinia polysora, à helmintosporiose (Exserohilum turcicum e à mancha branca(Phaeosphaeria maydis/Pantoea ananas. O experimento foi conduzido em blocos completos, com tratamentos ao acaso com três repetições, em Maringá-PR, na safra 2006/2007. Os tratamentos foram 27 híbridos de linhagens S5 e as cultivares BRS Angela, IAC 112 e Jade. Constataram-se diferenças significativas (p ≤ 0,05 entre os híbridos para todas as características avaliadas. O híbrido proveniente da combinação das linhagens C e K expressou os melhores índices de rendimento de grãos e capacidade de expansão e foi susceptível à ferrugem polissora. Os híbridos provenientes das linhagens L, H e Kdestacaram-se com relação à resistência à ferrugem polissora, enquanto os híbridos da linhagem G foram mais resistentes à helmintosporiose, e os das linhagens M e G, à mancha branca.In maize, variations on agronomic performance in distinctenvironments are common and more evident for one-way hybrids. The objective of this work was to evaluate the performance of new one-way popcorn hybrids developed by the Breeding Program of the State University of Maringá, in northwestern Paraná. The characteristics evaluated were: grain yield, popping expansion, plant height, ear height and the intensity of Southern rust (Puccinia polysora, Northern leaf blight (Exserohilum turcicum, and phaeosphaeria leaf spot (Phaeosphaeria maydis/Pantoea ananas. The

  2. Analysis of Potential Utilization and Disease-resistance of Maize Germplasm "Shen137"%玉米种质沈137抗病性及其利用潜力分析

    Institute of Scientific and Technical Information of China (English)

    甄广田; 罗玉鑫; 张振平

    2011-01-01

    在P群玉米种质沈137抗病鉴定结果的基础上,系统评价了沈137高抗弯孢叶斑病、灰斑病,兼抗大斑病、小斑病、南方锈病、穗腐病、茎腐病、丝黑穗病和矮花叶病等9种病害的抗病性及其抗病基因丰富而稳定的遗传特性.沈137是国内骨干自交系中抗病种类最多的自交系之一.充分利用沈137这类多抗资源及深入挖掘其应用潜力将成为应对玉米病害突发所带来生产隐患的重要途径.%Based on systemic identification of disease resistance of "Shenl37" ,one inbred line of the P group of maize germplasm,it was found that "Shenl37" is a good multiple resistance inbred line,with high resistance to Cur-vularia leaf blight and gray leaf spot, moderate resistance to Turcicum leaf blight, Maydis leaf blight, southern corn rust,ear rot,stalk rot,head smut,and dwarf mosaic virus,suggesting that the use of "Shenl37" is important to prevent the outbreak of disease problems in maize production.

  3. Isolation and antifungi mechanismanalysis of some marine actinomyces with antifungal activity.%抗真菌海洋放线菌的分离筛选与抗菌机制研究

    Institute of Scientific and Technical Information of China (English)

    马桂珍; 暴增海; 浦寅芳

    2009-01-01

    从江苏连云港海域采集海水、海泥、漂浮物、海洋动物样品,分离得到15株海洋放线菌.采用平板对峙培养法和打孔法测定放线菌不同菌株对玉米小斑病菌(Bipolaria maydis)、玉米圆斑病菌(Helminthosporium carbonum)、小麦赤霉病菌(Fusarium graminearum)、棉花枯萎病菌(Fusarium oxysporum f. sp. vasinfectum)、雪腐镰刀病菌(Fusarium nivale)、斑点落叶病菌(Alternaria alternata)、小麦根腐病菌(Bipolaris sorokiniana)、细链格孢病菌(Alternaria tenuis)、番茄早疫病菌(Alternaria solani)等植物病原真菌的抑制作用.结果表明:放线菌菌株BM-2、T-6、T-1-1、D-3、XS-X5-3、M-7、X-7对供试的植物病原真菌具有一定的抑制作用.其中BM-2菌株的抑菌作用最强,其发酵液能明显抑制多种植物病原真菌菌丝的生长,同时对细链格孢菌的分生孢子萌发和芽管伸长都有一定的抑制作用.

  4. Cytogenetic Mapping of Disease Resistance Genes and Analysis of Their Distribution Features on Chromosomes in Maize

    Institute of Scientific and Technical Information of China (English)

    Li Li-jia; Song Yun-chun

    2003-01-01

    Cytogenetic maps of four clusters of disease resistance genes were generated by ISH of the two RFLP markers tightly linked to and flanking each of maize resistance genes and the cloned resistance genes from other plant species onto maize chromosomes, combining with data published before. These genes include Helminthosporium turcium Pass resistance genes Ht1, Htn1 and Ht2, Helminthosporium maydis Nisik resistance genes Rhm1 and Rhm2, maize dwarf mosaic virus resistance gene Mdm1, wheat streak mosaic virus resistance gene Wsm1, Helminthosporium carbonum ULLstrup resistance gene Hml and the cloned Xanthomonas oryzae pv. Oryzae resistance gene Xa21 of rice, Cladosporium fulvum resistance genes Cf-9 and Cf-2.1 of tomato,and Pseudomonas syringae resistance gene RPS2 of Arabidopsis. Most of the tested disease resistance genes located on the four chromosomes, i.e., chromosomes1, 3, 6 and 8, and they closely distributed at the interstitial regions of these chromosomal long arms with percentage distances ranging 31.44(±3.72)-72.40(±3.25) except for genes Rhm1, Rhm2, Mdm1 and Wsm1 which mapped on the satellites of the short arms of chromosome6. It showed that the tested RFLP markers and genes were duplicated or triplicated in maize genome. Homology and conservation of disease resistance genes among species, and relationship between distribution features and functions of the genes were discussed. The results provide important scientific basis for deeply understanding structure and function of disease resistance genes and breeding in maize.

  5. Traditionally used medicinal plants against uncomplicated urinary tract infections: Are unusual, flavan-4-ol- and derhamnosylmaysin derivatives responsible for the antiadhesive activity of extracts obtained from stigmata of Zea mays L. against uropathogenic E. coli and Benzethonium chloride as frequent contaminant faking potential antibacterial activities?

    Science.gov (United States)

    Rafsanjany, Nasli; Sendker, Jandirk; Lechtenberg, Matthias; Petereit, Frank; Scharf, Birte; Hensel, Andreas

    2015-09-01

    The dried stigmata from Zea mays L. are used traditionally for the treatment of uncomplicated urinary tract infections. A recent screening has indicated that hydroalcoholic extract of the herbal material inhibits the adhesion of uropathogenic Escherichia coli (UPEC) to T24 bladder cells. For verification of these data EtOH-water (1:1) extracts from 4 different batches of Maydis stigmata were investigated. Within an in vitro adhesion assay (UPEC strain 2980 and human T24 bladder cells) a dose-dependent antiadhesive activity against UPEC was verified (IC50 1040μg/mL). Bioassay guided fractionation of M. stigmata, batch S1, by EtOH-water extraction, followed by chromatography on Sephadex LH20 revealed two active fractions (I and XI). Further purification of fraction I and structure elucidation of the isolated compound revealed the presence of significant amounts of the biocide benzethonium chloride as contaminant. Benzethonium chloride was also identified in subsequent investigations in 2 different batches of M. stigmata. The presence of such nondeclared and illegal contaminants in the herbal raw material market has to be discussed intensively. From benzethonium-free raw material (batch S2) as well as from batch S1 fraction XI was further fractionated by MPLC and preparative HPLC, leading to a still complex subfraction XIG, which was analyzed by UHPLC/+ESI-QTOF-MS analysis. Advanced data processing and species-metabolite relationship database revealed the tentatively existence of the unusual C-glycosidic flavones derhamnosylmaysin (6), 3'-deoxyrhamnosylmaysin (4), 3'-O-methylderhamnosylmaysin (3), apiferol (2) and alternanthin (8) which might be related to the antiadhesive activity of this subfraction against UPEC.

  6. Synthesis and Antifungal Activity of Novel 3-Caren-5-One Oxime Esters.

    Science.gov (United States)

    Huang, Min; Duan, Wen-Gui; Lin, Gui-Shan; Li, Kun; Hu, Qiong

    2017-09-12

    A series of novel 3-caren-5-one oxime esters were designed and synthesized by multi-step reactions in an attempt to develop potent antifungal agents. Two E-Z stereoisomers of the intermediate 3-caren-5-one oxime were separated by column chromatography for the first time. The structures of all the intermediates and target compounds were confirmed by UV-Vis, FTIR, NMR, ESI-MS, and elemental analysis. The antifungal activity of the target compounds was preliminarily evaluated by the in vitro method against Fusarium oxysporum f. sp. cucumerinum, Physalospora piricola, Alternaria solani, Cercospora arachidicola, Gibberella zeae,Rhizoeotnia solani, Bipolaris maydis, and Colleterichum orbicalare at 50 µg/mL. The target compounds exhibited best antifungal activity against P. piricola, in which compounds (Z)-4r (R = β-pyridyl), (Z)-4q (R = α-thienyl), (E)-4f' (R = p-F Ph), (Z)-4i (R = m-Me Ph), (Z)-4j (R = p-Me Ph), and (Z)-4p (R = α-furyl) had inhibition rates of 97.1%, 87.4%, 87.4%, 85.0%, 81.9%, and 77.7%, respectively, showing better antifungal activity than that of the commercial fungicide chlorothanil. Also, compound (Z)-4r (R = β-pyridyl) displayed remarkable antifungal activity against all the tested fungi, with inhibition rates of 76.7%, 82.7%, 97.1%, 66.3%, 74.7%, 93.9%, 76.7% and 93.3%, respectively, showing better or comparable antifungal activity than that of the commercial fungicide chlorothanil. Besides, the E-Z isomers of the target oxime esters were found to show obvious differences in antifungal activity. These results provide an encouraging framework that could lead to the development of potent novel antifungal agents.

  7. Aptitud combinatoria de lineas de maíz tropical con diferente capacidad para tolerar el achaparramiento

    Directory of Open Access Journals (Sweden)

    Guillermo Casta\\u00F1\\u00F3n

    2000-01-01

    Full Text Available Aptitud combinatoria de líneas de maíz tropical con diferente capacidad para tolerar el achaparramiento. En el presente trabajo, se evaluó en maíz las F1´s y los siete padres de éstas, para estudiar la respuesta de los híbridos y los padres al efecto del achaparramiento trasmitido por la chicharrita (Dalbulus maydis. Para ello se aplicó el Método 2, modelo I de Griffing (1956. De los resultados obtenidos, se encontró como más importantes a los efectos aditivos (ACG, que a los no-aditivos (ACE. Así también, se observó que los mejores cruzamientos para rendimiento de grano (RG, resultaron ser aquellas cruzas donde participaron una línea resistente (R x una susceptible (S. Las cruzas de menor RG, fueron entre progenitores R x R o bien S x S. Las líneas tres y cinco fueron las que presentaron los valores más altos de ACG para el carácter rendimiento de grano. Las estimas de las ACE, jugaron un papel importante en la estructura genética de los híbridos. La línea dos fue la que en combinaciones híbridas arrojó los valores más altos para ACE. Los progenitores uno y siete, fueron los peores en cuanto a ACG

  8. Research on the Isolation Method of Single Spore of Most Plant Pathogenic Fungi%一种适用于多数植物病原真菌的单孢分离方法

    Institute of Scientific and Technical Information of China (English)

    邱小燕; 汤智鹏; 张敏; 荆海瑜

    2011-01-01

    [目的]研究一种可靠并广泛适用于各种病原真菌的单孢分离技术.[方法]以玉米小斑病菌的单孢分离为例,并综合改进其他单孢分离方法,通过组织分离、孢悬液涂布、挑针转移3个步骤,挑取分散在清水琼脂平板上已经萌发的孢子获得单孢.[结果]采用该法对多种病害标本进行单孢分离,成功率都在92%以上,部分达到100%,单个菌丝片段的分离成功率达到60%以上.[结论]多种真菌的分离表明,该法操作简便,材料设备简单,结果可靠,成功率高,适用性广.%[ Objective ] The isolation technique of single spore, which was suitable for various pathogenic fungi, was researched. [ Method ] The isolation technique of single spore of Bipolaria maydis was improved and the single spore, which had germinated and scattered in the water agar plate, was picked up after three steps: tissue isolation, coating spore suspension and removal of dissecting needle. [ Result] The single spore of most of samples was isolated with the technique, which successful rate was over 92% and partially 100%; the successful rate of the isolation of single hyphal fragment was over 60%. [ Conclusion] The experimental result showed that the method was easy to operation, the requirement to material and equipment was simple, the result was reliable, the successful rate was high and the approach of its applicability was wide.

  9. Proteomics Associated with Virulence Differentiation of Curvularia lunata in Maize in China

    Institute of Scientific and Technical Information of China (English)

    Shufa Xu; Jie Chen; Lixing Liu; Xiaofei Wang; Xiuli Huang; Yuhong Zhai

    2007-01-01

    One-dimensional electrophoresis (1-DE) of proteins, two-dimensional electrophoresis (2-DE) of proteins and cloning of cDNA sequence were used to study the virulence differentiation of Curvularia lunata (Wakker) Boed. isolated from maize (Zea maydis L.) in China. From 1-DE gel profiles of proteins, 110 reproducible bands were separated from six isolates of C. lunata CX-3, SD-6, C-152, C107-1, DD-60 and W-18. Sixty-eight bands (61.82%) were polymorphic,suggesting huge biodiversities among the isolates. All isolates for the experiment were clustered into three groups consisting of different virulent types by coefficient value of 0.605. Group 1, consisting of CX-3, SD-6 and C-152 with high virulence displayed more protein bands than Groups 2 and 3, consisting of C107-1 and DD-60 with low virulence. Proteomics approaches based on 2-DE techniques were applied to identify specific proteins associated with the virulence differentiation in CX-3 and DD-60. A total of 423 protein spots were separated. Out of them 75 specific protein spots were displayed in 2-DE gels. Among them 28 protein spots were unique in CX-3 and eight in DD-60, and 39 protein spots were shown on both 2-DE gels but expressed differently in intensity. Twenty protein spots including three unique protein spots and 17 differentially expressed protein spots (more than two-fold DD-60) in CX-3 were further identified with MALDI-TOF MS/MS. Results indicated that most of the identified proteins were found to be associated with virulence differentiation, metabolisms, stress response and signal transduction.One of them was identified as Brn1 protein, which had been reported to be related to melanin biosynthesis and the virulence differentiation in fungi. Combined with our previous findings, we assumed that Brn1 protein and its regulating products might be involved in the virulence differentiation of C. lunata. Consequently, we cloned a Brn1 cDNA fragment and aligned it with the fragments in other fungi. Results

  10. Herencia de la resistencia al complejo Cercospora spp. en líneas de maíz tropical

    Directory of Open Access Journals (Sweden)

    Narro L. Luis

    2005-09-01

    collected suggested that maternal or cytoplasmic effects are important in controlling disease resistance. Accumulation of resistance genes through the development of synthetic varieties is an alternative to be implemented in maize breeding programs workin towards the development of gray leaf spot resistant tropical maize germplasm. Key words: Zea mays, Cercospora zeae maydis, gray leaf spot of maize, diallel mating, general combining ability, specific combining ability, reciprocal effects.

  11. 玉米灰斑病及抗性研究%The Research Advance on Resistance to Grey Leaf Spot in Maize

    Institute of Scientific and Technical Information of China (English)

    曹国辉

    2009-01-01

    玉米灰斑病由尾孢菌引起.尾孢菌在玉米叶粉碳酸钙琼脂培养基上能够产生分生孢子.采用注射法可以进行人工接种,空气湿度大、温度适宜条件下利于病害发生.参考玉米大、小斑病国家调查标准,提出9级标准记载植株灰斑病发病级别.研究表明,以推广种植抗病品种为主,辅助以栽培管理的综合防治措施是防治玉米灰斑病的有效途径.目前,在我国主要玉米种质中,仅有PB类群种质表现为抗灰斑病,四平头、旅大红骨、PA等类群种质多表现为感病.随着玉米灰斑病危害加重,应加强玉米灰斑病抗性基因发掘、种质改良和创新研究.%Grey leaf spot(GLS) caused by Gercospora zeae-maydis is one of the important diseases in maize worldwide. In recent years, GLS occurs more and more severely and becomes the major disease in maize growing area of China. GLS can produce conidia on the medium of maize leaf powder plus CaCO3 agar and can be artificially inoculated by injection method. High humidity and suitable temperature are very conducive for the occurrence of GLS. According to the national evaluation standard of maize exserohilum turcicum and bipolarize maydays, 9 scales is suggest-ed for evaluating GLS severity. Development and cultivation of GLS resistant hybrids is the effective way to control GLS. Currently, PB germplasm is identified to be resistant to GLS, while Sipingtou, Lvdahonggu and PA germplasm are most susceptible. With GLS occurs more widely in China, gene identification and utilization and germplasm improvement for GLS resistance should be strengthened.

  12. Fungal Endophyte Diversity and Bioactivity in the Indian Medicinal Plant Ocimum sanctum Linn.

    Directory of Open Access Journals (Sweden)

    Kanika Chowdhary

    Full Text Available Endophytic mycopopulation isolated from India's Queen of herbs Tulsi (Ocimum sanctum were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues from three different geographic locations (Delhi, Hyderabad and Mukteshwar during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011 in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907 whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846. Mukteshwar (altitude: 7500 feet reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in

  13. Quimio e termoterapia em sementes e aplicação de fungicidas em Brachiaria brizantha como estratégias no manejo do carvão Chemical and thermal therapy in seeds and application of fungicides in Brachiaria brizantha as strategies for managing smut

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Marchi

    2008-12-01

    Full Text Available Objetivou-se estudar os potenciais dos tratamentos térmico e químico em sementes de Brachiaria brizantha cv. Piatã, preconizados como eficientes na superação da dormência física das mesmas, na redução do inóculo de carvão (Ustilago operta. Amostras de sementes foram expostas ao ácido sulfúrico por tempo variando de 0 a 10 min, ou imersas em água à temperatura variável (27 a 60 ºC por 5 ou 10 min. Avaliaram-se os efeitos dos tratamentos no número de teliósporos de U. operta remanescentes e na freqüência de germinação das sementes tratadas. À medida que se prolongou a escarificação química das sementes se evidenciou decréscimo do inóculo de carvão. Menor número de teliósporos foi observado nas sementes submetidas à quimioterapia por 10 min. Em geral, a exposição das sementes ao H2SO4 não levou ao decréscimo da capacidade germinativa das mesmas. Os resultados da termoterapia não foram tão evidentes como os observados para a quimioterapia, sejam na redução do número de teliósporos ou na superação da dormência. É provável que o tempo de exposição das sementes não tenha sido o suficiente. Paralelamente, verificou-se a eficiência da aplicação de fungicidas na parte aérea das plantas no controle da incidência do carvão. Foram testados os fungicidas: 1 - tiofanato metílico + chlorothalonil; 2 - mancozeb; 3 - tebuconazole; 4 - triadimenol; 5 - azoxystrobin + cyproconazole; 6 - pyraclostrobin + epoxyconazole; 7 - trifloxystrobin + cyproconazole; 8 - carboxin + thiram; 9 - tiofanato metílico + flutriafol; 10 - carbendazin, e 11 - difenoconazole. Mesmo com a ocorrência de baixa incidência de carvão na área experimental (máximo de 15%, observaram-se diferenças entre os fungicidas quanto ao controle da doença. O fungicida tebuconazole e as formulações pyraclostrobin + epoxyconazole e trifloxystrobin + cyproconazole foram os mais promissores.This work investigated the potentials of the thermal

  14. Effects of Different Altitudes on Growth and Culm-swelling of Zizania caduci-fl ora(Turcz.)Hand.-Mazz.%不同海拔高度对茭白生长及孕茭的影响

    Institute of Scientific and Technical Information of China (English)

    邓建平; 石敏; 黄建中; 郭得平; 胡振亮

    2013-01-01

    The present study investigates the effects of high altitudes (200-1 000 m) on the growth and culm-swelling of Zizania caduciflora(Turcz.)Hand. -Mazz..The results showed that the sprout-ing date,leafing and peak value of plant height increase at high altitudes were significantly delayed and lower than those grown at lower altitudes.Sprouting of Zizania caduciflora(Turcz.)Hand. -Mazz. in high altitudes requires over 5 °C effective accumulated temperature than in low altitudes. Its leafing rate, plant height increasing speed showed positive correlation with the effective accumulated temperature in various altitudes.Culm-swelling was observed to take place in the following order:410 m> 650 m> 1 008 m > 815 m.The culm-swelling stage usually occurred during the period,when the tempera-ture was suitable.Therefore,the effect of altitude on growth and culm-swelling of Zizania caduciflora (Turcz.)Hand.-Mazz. was mainly realized by temperature differences at different altitudes.Suppres-sion of growth and metabolism of Ustilago esculenta by high temperature is most likely to be the limiting factor for culm-swelling of Zizania caduciflora(Turcz.)Hand. -Mazz. during hot summer days.%  对在海拔高度200~1000 m地区栽培的单季茭的生长和孕茭特性进行研究。结果表明:高海拔地区栽培的茭白萌芽期、出叶和株高增速峰值均比低海拔处明显延迟。高海拔处茭白萌芽较低海拔处需要更多5℃以上的有效积温;各海拔处茭白出叶速率和株高增速与有效积温间呈正相关。各海拔高度孕茭先后顺序为:410 m>650 m>1008 m>815 m。各海拔处茭白的孕茭期均发生在气温适宜孕茭的时间段内。因此,海拔高度对茭白生长和孕茭的影响主要是通过不同海拔高度的温度差异来实现的,高温抑制茭白黑粉菌的正常生长及代谢可能是茭白夏季高温条件下难以孕茭的重要原因。

  15. 枯草芽孢杆菌B47菌株高产抗菌物质的培养基及发酵条件优化%Optimization of culture medium and fermentation conditions for high production of antimicrobial substance by Bacillus subtilis strain B47

    Institute of Scientific and Technical Information of China (English)

    叶云峰; 黎起秦; 袁高庆; 付岗; 缪剑华; 林纬

    2011-01-01

    Bacillus subtitis strain B47 is an endophytic bacterium of tomato and can produce substance to inhibit the growth of Bipolaris maydis which can cause southern corn leaf blight. The optimal nitro-gen source, carbon source and salt for the production of antimicrobial substance by strain B47 were tested. The medium composition and fermentation conditions were optimized by orthogonal experiments. The results showed that the optimal nitrogen source, carbon source and salt were yeast extract; sucrose and MgSO4-7H2O, respectively. The best medium was YSB (Yeast extract-sucrose-beef extract). The composition of the medium was 2% {W/V) sucrose, 2% {W/V) yeast extract, 1.5% {W/V) beef extract, 0.06% {W/V) MgSO4-7H2O and 0.000 9% {W/V) FeSO4-7H2O. The optimal fermentation conditions were the combination of temperature 30 °C, initial pH 7.0, incubation time 6 d, 1% inoculum volume percentage and medium volume 40 mL/200 mL.%枯草芽孢杆菌Bacillus subtilis B47菌株为番茄内生细菌,也是玉米小斑病拮抗茵,能产生对玉米小斑病菌有强烈抑制作用的抗菌物质.以B47菌株发酵液的无菌滤液对玉米小斑病菌的抗茵活性为检测指标,测定B47菌株产抗菌物质培养所需的最佳碳,氮源和无机盐,并通过正交试验法对该菌株产抗茵物质的培养基配方和摇瓶发酵条件进行优化.研究结果表明,B47菌株产抗菌物质最佳碳、氮源和无机盐分别为蔗糖、酵母浸膏和MgSO4·7H2O,最优培养基是YSB (Yeast extract-sucrose-beef extract)培养基,其配方为:蔗糖2%,酵母浸膏2%,牛肉浸膏1.5%,MgSO4·7H2O0.06%,FeSO4·7H2O 0.000 9%,最优发酵条件组合为:30℃,pH 7.0,170 r/min摇床培养6d,接种量为1%,装液量为40mL/200mL.

  16. 珠芽蓼内生菌Z17抑菌能力测定及其鉴定%Identification of Polygonum viviparum endophytic bacteria Z17 and its capacity to antagonistic towards pathogenic fungi

    Institute of Scientific and Technical Information of China (English)

    李振东; 陈秀蓉; 杨成德

    2011-01-01

    以高寒草地优势植物珠芽蓼(Polygonum viviparum)的一内生细菌菌株Z17为研究对象,采用平板对峙法测定其对7种植物病原真菌的抑菌能力,利用形态学和分子生物学2种方法确定其分类地位。该菌对玉米小斑病菌(Bipolaria maydis)、立枯丝核病菌(Rhizoctonia solani)、菌核病菌(Sclerotinia sclerotiorum)、西瓜尖镰孢(Fusarium oxysporumf.Niveum)、番茄早疫病菌(Alternaria solani)、番茄灰霉病菌(Botrytis cinerea)和小麦离蠕孢菌(Bipolaris sorokiniana)等均有抑制作用,抑菌谱较广。菌体杆状,菌体大小为(1.4~3.6)μm×(0.4~0.6)μm,革兰氏阳性,中生芽孢,与芽孢杆菌的形态一致;16SrDNA基因序列与莫海威芽孢杆菌(Bacillus mo-javensis)的模式菌株BCRC 17531相似度达99.79%,在系统发育树上Z17与菌株BCRC 17531的遗传距离小于0.000 5,故鉴定为莫海威芽孢杆菌。该菌可能具有开发生物农药的潜力。%Polygonum viviparum is the dominant species of alpine meadow community.A experiment was conducted to estimate the capacity of P.viviparum endophytic bacteria Z17 to antagonistic towards pathogenic fungi by PDA plates and to determine the taxonomic status of Z17 by morphological and molecular biology methods.The results of this study showed that Z17 inhibited Bipolaria maydis,Rhizoctonia solani,Sclerotinia sclerotiorum,Fusarium oxysporum f.Niveum,Alternaria solani,Botrytis cinerea,Bipolaris sorokiniana,indicating that Z17 had a wide antibacterial spectrum.Z17 was rod shape with a size of(1.4-3.6) μm ×(0.4-0.6) μm,Gram-positive,and produced spore in the center,and it was same morphological characteristics with Bacillus sp.Similarity between 16S rDNA gene sequence of Z17 and Bacillus mojavensis model strain BCRC 17531 reached 99.79%,and the genetic distance between Z17 and BCRC 17531 in the phylogenetic tree was less than 0.000 5.These results suggested that Z17 could be

  17. Study on the Pharmacodynamic of Ethanol Extracts from Lonicera japonica in Rats with Hepatic Fibrosis%玉米须醇提物对肝纤维化大鼠药效学研究

    Institute of Scientific and Technical Information of China (English)

    陈艳军; 高旭珍; 关大勇; 耿丹; 张敬华

    2012-01-01

    ).%目的:观察玉米须(stigma maydis or com si]k)醇提取物对实验性肝纤维化大鼠的治疗效果,并探讨其作用机制.方法:用四氯化碳( CCl4)诱导大鼠肝纤维化模型(皮下注射为8周,腹腔注射为4周),将实验动物随机分为正常对照组(A)、模型组(B)、秋水仙碱组(C,0.2 mg·kg -1·d-1)、玉米须醇提取物组(D,5.4g·kg-1·d-1).除正常对照组外,其余3组均用四氯化碳(sc,每次3 mL·kg-1,2次/周)诱发肝纤维化.各组于造模第8周末处死动物,分别用放射免疫法检测血清层黏连蛋白(LN),m型前胶原( PCⅢ),透明质酸(HA)及Ⅳ型胶原(CⅣ);RT-PCR检测肝组织Smad3 mRNA的表达;作HE染色和胶原纤维特殊染色行肝组织病理形态学观察及电镜观察治疗前后大鼠肝脏的超微结构.结果:模型组与正常组比较,大鼠血清中LN(207.6±27.4)μg·L-1,PCⅢ(280.1±3.2) μg· L-1,HA(357.0±14.1) μg·L-1,CⅣ(180.0±7.0) μg·L-1,大鼠肝组织内Smad,mRNA(0.865±0.084)水平明显升高(P<0.01),经玉米须醇提取物治疗8周,大鼠血清中LN(162.4±2.2) μg·L-1,PCⅢ(193.2±1.8)μg·L-1,HA(219.4±3.7) μg· L-1,CⅣ( 138.0±2.5)μg·L-1水平明显降低(P<0.01);大鼠肝组织内Smad3 mRNA(0.410±0.026)表达明显下降(P<0.01);大鼠肝组织病理学检测改善显著.结论:玉米须能有效地抑制肝纤维化的发展,其机制可能是通过下调肝组织内Smad,mRNA表达,降低ECM的分泌而达到的.