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Lauren A Ostrowski, Barry J Saville
The basidiomycete smut fungus Ustilago maydis causes common smut of corn. This disease is spread through the production of teliospores, which are thick-walled dormant structures characterized by low rates of respiration and metabolism. Teliospores are formed when the fungus grows within the plant, and the morphological steps involved in their formation have been described, but the molecular events leading to dormancy are not known. In U. maydis, natural antisense transcripts (NATs) can function to alter gene expression and many NATs have increased levels in the teliospore...
November 26, 2016: Molecular Microbiology
Franziska Rabe, Jason Bosch, Alexandra Stirnberg, Tilo Guse, Lisa Bauer, Denise Seitner, Fernando A Rabanal, Angelika Czedik-Eysenberg, Simon Uhse, Janos Bindics, Bianca Genenncher, Fernando Navarrete, Ronny Kellner, Heinz Ekker, Jochen Kumlehn, John P Vogel, Sean P Gordon, Thierry C Marcel, Martin Münsterkötter, Mathias C Walter, Christian Mk Sieber, Gertrud Mannhaupt, Ulrich Güldener, Regine Kahmann, Armin Djamei
Due to their economic relevance, the study of plant pathogen interactions is of importance. However, elucidating these interactions and their underlying molecular mechanisms remains challenging since both host and pathogen need to be fully genetically accessible organisms. Here we present milestones in the establishment of a new biotrophic model pathosystem: Ustilago bromivora and Brachypodium sp. We provide a complete toolset, including an annotated fungal genome and methods for genetic manipulation of the fungus and its host plant...
November 11, 2016: ELife
Gero Steinberg
Peroxisomes (POs) are an essential part of the fungal cell's inventory. They are involved in cellular lipid homeostasis, reactive oxygen metabolism and in the synthesis of secondary metabolites. These functions are thought to require frequent organelle-organelle interactions and the even-distribution of POs in fungal hypha. Recent work in the basidiomycete Ustilago maydis and the ascomycete Aspergillus nidulans reveals a crucial role of early endosomes (EEs) in the dynamic behavior of POs required for their cellular distribution and interaction...
December 2016: Fungal Genetics and Biology: FG & B
Brett J Green, Angela R Lemons, Yeonmi Park, Jean M Cox-Ganser, Ju-Hyeong Park
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)...
October 27, 2016: Journal of Occupational and Environmental Hygiene
Kristin Bösch, Lamprinos Frantzeskakis, Miroslav Vraneš, Jörg Kämper, Kerstin Schipper, Vera Göhre
Gene deletion plays an important role in the analysis of gene function. One of the most efficient methods to disrupt genes in a targeted manner is the replacement of the entire gene with a selectable marker via homologous recombination. During homologous recombination, exchange of DNA takes place between sequences with high similarity. Therefore, linear genomic sequences flanking a target gene can be used to specifically direct a selectable marker to the desired integration site. Blunt ends of the deletion construct activate the cell's DNA repair systems and thereby promote integration of the construct either via homologous recombination or by non-homologous-end-joining...
September 30, 2016: Journal of Visualized Experiments: JoVE
Elena Geiser, Sandra K Przybilla, Meike Engel, Wiebke Kleineberg, Linda Büttner, Eda Sarikaya, Tim den Hartog, Jürgen Klankermayer, Walter Leitner, Michael Bölker, Lars M Blank, Nick Wierckx
The Ustilaginaceae family of smut fungi, especially Ustilago maydis, gained biotechnological interest over the last years, amongst others due to its ability to naturally produce the versatile bio-based building block itaconate. Along with itaconate, U. maydis also produces 2-hydroxyparaconate. The latter was proposed to be derived from itaconate, but the underlying biochemistry and associated genes were thus far unknown. Here, we confirm that 2-hydroxyparaconate is a secondary metabolite of U. maydis and propose an extension of U...
October 14, 2016: Metabolic Engineering
Lucia Rochi, María José Diéguez, Germán Burguener, Martín Alejandro Darino, María Fernanda Pergolesi, Lorena Romina Ingala, Alba Romina Cuyeu, Adrián Turjanski, Enrique Domingo Kreff, Francisco Sacco
Rust fungi are one of the most devastating pathogens of crop plants. The biotrophic fungus Puccinia sorghi Schwein (Ps) is responsible for maize common rust, an endemic disease of maize (Zea mays L) in Argentina that causes significant yield losses in corn production. In spite of this, the Ps genomic sequence was not available. We used Illumina sequencing to rapidly produce the 99.6 Mb draft genome sequence of Ps race RO10H11247, derived from a single-uredinial isolate from infected maize leaves collected in the Argentine Corn Belt Region during 2010...
October 13, 2016: Fungal Genetics and Biology: FG & B
María F Jiménez-Becerril, Sanjuana Hernández-Delgado, Myrna Solís-Oba, Juan M González Prieto
The current understanding of the genetic diversity of the phytopathogenic fungus Ustilago maydis is limited. To determine the genetic diversity and structure of U. maydis, 48 fungal isolates were analyzed using mitochondrial simple sequence repeats (SSRs). Tumours (corn smut or 'huitlacoche') were collected from different Mexican states with diverse environmental conditions. Using bioinformatic tools, five microsatellites were identified within intergenic regions of the U. maydis mitochondrial genome. SSRMUM4 was the most polymorphic marker...
October 11, 2016: Mitochondrial DNA. Part A. DNA Mapping, Sequencing, and Analysis
Libera Lo Presti, Bernd Zechmann, Jochen Kumlehn, Liang Liang, Daniel Lanver, Shigeyuki Tanaka, Ralph Bock, Regine Kahmann
Successful colonization of plants by prokaryotic and eukaryotic pathogens requires active effector-mediated suppression of defense responses and host tissue reprogramming. Secreted effector proteins can either display their activity in the apoplast or translocate into host cells and function therein. Although characterized in bacteria, the molecular mechanisms of effector delivery by fungal phytopathogens remain elusive. Here we report the establishment of an assay that is based on biotinylation of effectors in the host cytoplasm as hallmark of uptake...
September 26, 2016: New Phytologist
Kevin Felipe Cruz Martho, Amanda Teixeira de Melo, Juliana Possato Fernandes Takahashi, Juliana Mariotti Guerra, Dayane Cristina da Silva Santos, Sônia Ueda Purisco, Márcia de Souza Carvalho Melhem, Raquel Dos Anjos Fazioli, Clerlune Phanord, Patrícia Sartorelli, Marcelo A Vallim, Renata C Pascon
Fungal opportunistic pathogens colonize various environments, from plants and wood to human and animal tissue. Regarding human pathogens, one great challenge during contrasting niche occupation is the adaptation to different conditions, such as temperature, osmolarity, salinity, pressure, oxidative stress and nutritional availability, which may constitute sources of stress that need to be tolerated and overcome. As an opportunistic pathogen, C. neoformans faces exactly these situations during the transition from the environment to the human host, encountering nutritional constraints...
2016: PloS One
Janpeter Stock, Marius Terfrüchte, Kerstin Schipper
Unconventional secretion of proteins in eukaryotes is characterized by the circumvention of the Endoplasmic Reticulum (ER). As a consequence proteins exported by unconventional pathways lack N-glycosylation, a post-transcriptional modification that is initiated in the ER during classical secretion. We are exploiting the well-established enzyme β-glucuronidase (GUS) to assay unconventional protein secretion (UPS). This bacterial protein is perfectly suited for this purpose because it carries a eukaryotic N-glycosylation motif...
2016: Methods in Molecular Biology
Maribel Valdez-Morales, L Céspedes Carlos, María Elena Valverde, Enrique Ramírez-Chávez, Octavio Paredes-López
Huitlacoche mushroom (composed by the fruiting bodies growing on the maize ears from the basidiomycete Ustilago maydis) is a culinary delicacy with a great economic and nutraceutical value. In this work, phenolic content, antioxidant activity, ergosterol and fatty acids profile from huitlacoche produced in 15 creole and in one hybrid maize genotypes, and harvested at different stages of development were determined. The hybrid crop was studied in raw and cooked samples. Total phenolic content ranged from 415...
September 7, 2016: Plant Foods for Human Nutrition
Mine Erdem, Zülal Kesmen, Esra Özbekar, Bülent Çetin, Hasan Yetim
A new method based on high resolution melting (HRM) analysis was developed for the differentiation and classification of the yeast species that cause food spoilage. A total 134 strains belonging to 21 different yeast species were examined to evaluate the discriminative power of HRM analysis. Two different highly variable DNA regions on the 26 rRNA gene were targeted to produce the HRM profiles of each strain. HRM-based grouping was compared and confirmed by (GTG)5 rep-PCR fingerprinting analysis. All of the yeast species belonging to the genera Pichia, Candida, Kazachstania, Kluyveromyces, Debaryomyces, Dekkera, Saccharomyces, Torulaspora, Ustilago, and Yarrowia, which were produced as species-specific HRM profiles, allowed discrimination at species and/or strain level...
September 2016: Journal of Microbiology / the Microbiological Society of Korea
Matthias Kretschmer, Daniel Croll, James W Kronstad
The ability of biotrophic fungi to metabolically adapt to the host environment is a critical factor in fungal diseases of crop plants. In this study, we analyzed the transcriptome of maize tumors induced by Ustilago maydis to identify key features underlying metabolic shifts during disease. Among other metabolic changes, this analysis highlighted modifications during infection in transcriptional regulation of carbohydrate allocation and starch metabolism. We confirmed the relevance of these changes by establishing that symptom development was altered in an id1 (indeterminate1) mutant that has increased accumulation of sucrose as well as being defective in the vegetative to reproductive transition...
August 26, 2016: Molecular Plant Pathology
Matthias Kretschmer, Daniel Croll, James W Kronstad
Biotrophic fungal pathogens must evade or suppress plant defence responses to establish a compatible interaction in living host tissue. In addition, metabolic changes during disease reflect both the impact of nutrient acquisition by the fungus to support proliferation and the integration of metabolism with the plant defence response. In this study, we used transcriptome analyses to predict that the chloroplast and associated functions are important for symptom formation by the biotrophic fungus Ustilago maydis on maize...
August 26, 2016: Molecular Plant Pathology
Martin Schuster, Magdalena Martin-Urdiroz, Yujiro Higuchi, Christian Hacker, Sreedhar Kilaru, Sarah J Gurr, Gero Steinberg
Fungal cells are surrounded by an extracellular cell wall. This complex matrix of proteins and polysaccharides protects against adverse stresses and determines the shape of fungal cells. The polysaccharides of the fungal wall include 1,3-β-glucan and chitin, which are synthesized by membrane-bound synthases at the growing cell tip. A hallmark of filamentous fungi is the class V chitin synthase, which carries a myosin-motor domain. In the corn smut fungus Ustilago maydis, the myosin-chitin synthase Mcs1 moves to the plasma membrane in secretory vesicles, being delivered by kinesin-1 and myosin-5...
2016: Nature Microbiology
Juan Antonio Cervantes-Montelongo, Elva Teresa Aréchiga-Carvajal, José Ruiz-Herrera
Fungi are capable to adapt to environments with different pH values. Here we used microarrays to analyze the transcriptomic response of the Basidiomycota Ustilago maydis when transferred from a neutral pH medium to acidic, or alkaline media. Yeast and hyphal monomorphic mutants were used as controls, permitting the identification of 301 genes differentially regulated during the transfer from neutral to an acidic medium, of which 162 were up-regulated and 139 down-regulated. When cells were transferred to an alkaline medium, we identified 797 differentially regulated genes, 335 up-regulated, and 462 down-regulated...
November 2016: Journal of Basic Microbiology
Alexandra Matei, Gunther Doehlemann
Ustilago maydis is a well-established model system for biotrophic fungal plant pathogens. The fungus has a dimorphic life cycle with a yeast-like saprophytic phase switching to filamentous, pathogenic growth upon hyphal fusion. Due to its highly differentiated development and the amenability for reverse-genetics U. maydis provides a model system for both fungal cell biology as well as the study of biotrophic plant interaction. The present article highlights key findings in different aspects of cell biology on the corn smut disease and provides an outlook on the most intriguing open questions...
August 6, 2016: Current Opinion in Microbiology
S Baumann, S Zander, S Weidtkamp-Peters, M Feldbrügge
Septins are highly conserved cytoskeletal proteins involved in a variety of biological processes such as cell polarization and cytokinesis. In humans, functional defects in these proteins have been linked to cancer and neuronal diseases. In recent years, substantial progress has been made in studying the structure of septin subunits and the formation of defined heteromeric building blocks. These are assembled into higher-order structures at distinct subcellular sites. An important microscopic approach in studying septin assembly and dynamics is the use of septins tagged with fluorescent proteins...
2016: Methods in Cell Biology
Thiemo Zambanini, Joerg M Buescher, Guido Meurer, Nick Wierckx, Lars M Blank
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.
2016: Genome Announcements
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