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Angelika Czedik-Eysenberg, Sebastian Seitner, Ulrich Güldener, Stefanie Koemeda, Jakub Jez, Martin Colombini, Armin Djamei
There is a need for flexible and affordable plant phenotyping solutions for basic research and plant breeding. We demonstrate our open source plant imaging and processing solution ('PhenoBox'/'PhenoPipe') and provide construction plans, source code and documentation to rebuild the system. Use of the PhenoBox is exemplified by studying infection of the model grass Brachypodium distachyon by the head smut fungus Ustilago bromivora, comparing phenotypic responses of maize to infection with a solopathogenic Ustilago maydis (corn smut) strain and effector deletion strains, and studying salt stress response in Nicotiana benthamiana...
April 5, 2018: New Phytologist
Yafen Zhang, Honglei Liu, Qianchao Cao, Qianwen Ge, Haifeng Cui, Xiaoping Yu, Zihong Ye
Ustilago esculenta, an obligate parasite of Zizania latifolia, is a typical dimorphic fungus, which induces host stem swelling and inhibits host inflorescence development, but is not found in host leaves. Previous studies have shown that dimorphic switching is essential for fungal pathogenicity and is regulated by PKA and mitogen-activated protein kinase (MAPK) signaling pathways that are integrated by Prf1 in Ustilago maydis. In this study, we identified a Prf1 homolog in U. esculenta, designated UePrf1, encoding 830 amino acids, with a conserved high mobility group (HMG) domain located between amino acids 124 and 195...
March 30, 2018: FEMS Microbiology Letters
Sakon Monggoot, Tanakrit Pichaitam, Chutima Tanapichatsakul, Patcharee Pripdeevech
Mitrephora wangii, an ethnomedicinal plant, has been used as a natural antibiotic and immunity booster in Thailand. A total of 22 fungi were isolated from M. wangii flowers. The fungal isolates were categorized into six genera including Agrocybe, Aspergillus, Colletotrichum, Nigrospora, Puccinia and Ustilago. Most extracts exhibited antibacterial activity against at least one of the test bacteria. Aspergillus sp. MFLUCC16-0845 was identified as the most bioactive fungus. Chemical composition of Aspergillus sp...
April 2, 2018: Archives of Microbiology
Yafen Zhang, Qianwen Ge, Qianchao Cao, Haifeng Cui, Peng Hu, Xiaoping Yu, Zihong Ye
Ustilago esculenta, resembling a fungal endophyte in Zizania latifolia, inhibits the host plant flowering and induces the host stems to swell and form edible galls. It is well believed that when and how the fungus infects and proliferates in the host plants during the host development is of importance in the edible gall formation. Mitogen-activated protein kinases (MAPKs) have been found to play an important role in sensing environment cues and regulating infection. Two MAPK genes UeKpp2 and UeKpp6 from U. esculenta were cloned and suggested to be involved in the Fus3/Kss1 pathway by a phylogenetic analysis with the neighbor-joining method...
March 28, 2018: Current Microbiology
David Schuler, Christina Höll, Nathalie Grün, Jonas Ulrich, Bastian Dillner, Franz Klebl, Alexandra Ammon, Lars M Voll, Jörg Kämper
In most organisms, galactose is metabolized via the Leloir pathway, which is conserved from bacteria to mammals. Utilization of galactose requires a close interplay of the metabolic enzymes, as misregulation or malfunction of individual components can lead to the accumulation of toxic intermediate compounds. For the phytopathogenic basidiomycete Ustilago maydis, galactose is toxic for wildtype strains, i.e. leads to growth repression despite the presence of favorable carbon sources as sucrose. The galactose sensitivity can be relieved by two independent modifications: (1) by disruption of Hxt1, which we identify as the major transporter for galactose, and (2) by a point mutation in the gene encoding the galactokinase Gal1, the first enzyme of the Leloir pathway...
March 23, 2018: Fungal Genetics and Biology: FG & B
Vikas Yadav, Sheng Sun, R Blake Billmyre, Bhagya C Thimmappa, Terrance Shea, Robert Lintner, Guus Bakkeren, Christina A Cuomo, Joseph Heitman, Kaustuv Sanyal
The centromere DNA locus on a eukaryotic chromosome facilitates faithful chromosome segregation. Despite performing such a conserved function, centromere DNA sequence as well as the organization of sequence elements is rapidly evolving in all forms of eukaryotes. The driving force that facilitates centromere evolution remains an enigma. Here, we studied the evolution of centromeres in closely related species in the fungal phylum of Basidiomycota. Using ChIP-seq analysis of conserved inner kinetochore proteins, we identified centromeres in three closely related Cryptococcus species: two of which are RNAi-proficient, while the other lost functional RNAi...
March 5, 2018: Proceedings of the National Academy of Sciences of the United States of America
Sachin Kumar, Ron E Knox, Asheesh K Singh, Ron M DePauw, Heather L Campbell, Julio Isidro-Sanchez, Fran R Clarke, Curtis J Pozniak, Amidou N'Daye, Brad Meyer, Andrew Sharpe, Yuefeng Ruan, Richard D Cuthbert, Daryl Somers, George Fedak
Loose smut, caused by Ustilago tritici (Pers.) Rostr., is a systemic disease of tetraploid durum wheat (Triticum turgidum L.). Loose smut can be economically controlled by growing resistant varieties, making it important to find and deploy new sources of resistance. Blackbird, a variety of T. turgidum L. subsp. carthlicum (Nevski) A. Love & D. Love, carries a high level of resistance to loose smut. Blackbird was crossed with the loose smut susceptible durum cultivar Strongfield to produce a doubled haploid (DH) mapping population...
2018: PloS One
Sebastian Ziemann, Karina van der Linde, Urs Lahrmann, Beyda Acar, Farnusch Kaschani, Tom Colby, Markus Kaiser, Yezhang Ding, Eric Schmelz, Alisa Huffaker, Nicholas Holton, Cyril Zipfel, Gunther Doehlemann
Localized control of cell death is crucial for the resistance of plants to pathogens. Papain-like cysteine proteases (PLCPs) regulate plant defence to drive cell death and protection against biotrophic pathogens. In maize (Zea mays), PLCPs are crucial in the orchestration of salicylic acid (SA)-dependent defence signalling. Despite this central role in immunity, it remains unknown how PLCPs are activated, and which downstream signals they induce to trigger plant immunity. Here, we discover an immune signalling peptide, Z...
February 26, 2018: Nature Plants
Lu Zhou, Theresa Obhof, Karina Schneider, Michael Feldbrügge, G Ulrich Nienhaus, Jörg Kämper
In the phytopathogenic basidiomycete Ustilago maydis, the Num1 protein has a pivotal function in hyphal morphogenesis. Num1 functions as a core component of the spliceosome-associated Prp19/CDC5 complex (NTC). The interaction of Num1 with the kinesin motor Kin1 suggests a connection between a component of the splicing machinery and cytoplasmic trafficking processes. Previously it was shown that Num1 localizes predominantly in the nucleus; however, due to the diffraction-limited spatial resolution of conventional optical microscopy, it was not possible to attribute the localization to specific structures within the cytoplasm...
February 26, 2018: Scientific Reports
Patrick Feeney, Santiago Fernández Rodríguez, Rafael Molina, Eoin McGillicuddy, Stig Hellebust, Michael Quirke, Shane Daly, David O'Connor, John Sodeau
An air measurement campaign was carried out at a green-waste composting site in the South of Ireland during Spring 2016. The aim was to quantify and identify the levels of Primary Biological Aerosol Particles (PBAP) that were present using the traditional off-line, impaction/optical microscopy method alongside an on-line, spectroscopic approach termed WIBS (Wideband Integrated Bioaerosol Sensor), which can provide number concentrations, sizes and "shapes" of airborne PBAP in real-time by use of Light Induced Fluorescence (LIF)...
February 21, 2018: Waste Management
Wen Chen, Sarah Hambleton, Keith A Seifert, Odile Carisse, Moussa S Diarra, Rick D Peters, Christine Lowe, Julie T Chapados, C André Lévesque
Spore samplers are widely used in pathogen-surveillance but not so much for monitoring the composition of aeromycobiota. In Canada, a nationwide spore-sampling network (AeroNet) was established as a pilot project to assess fungal community composition in air and rain samples collected using three different spore samplers in the summers of 2010 and 2011. Metabarcodes of the Internal Transcribed Spacer (ITS) were exhaustively characterized for three of the network sites, in British Columbia (BC), Québec (QC) and Prince Edward Island (PEI), to compare performance of the samplers...
February 23, 2018: Applied and Environmental Microbiology
Qingwen Zhou, William K Holloman, Milorad Kojic
Primary components of the homologous recombination pathway in eukaryotes include Rad51 whose function is to search for DNA sequence homology and promote strand exchange, its mediator BRCA2, and Dss1, a key regulator of BRCA2. We seek to understand the role of BRCA2 in governing the activity of Rad51 and to learn how BRCA2 function is regulated by Dss1. We use the microbe Ustilago maydis as a model system for experimentation because it has a well-conserved BRCA2-homolog, Brh2, and is amenable to biochemical and molecular genetic manipulations and analysis...
2018: Methods in Enzymology
Karina Van Der Linde, Ljudmilla Timofejeva, Rachel L Egger, Birger Ilau, Reza Hammond, Chong Teng, Blake C Meyers, Gunther Doehlemann, Virginia Walbot
Small proteins are crucial signals during development, host defense, and physiology. Spatiotemporal restricted functions of signaling proteins remain challenging to study in planta. The several month span required to assess transgene expression, particularly in flowers, combined with the uncertainties from transgene position effects and ubiquitous or overexpression, makes monitoring of spatiotemporally restricted signaling proteins lengthy and difficult. This situation could be rectified with a transient assay in which protein deployment is controlled spatially and temporally in planta to assess protein functions, timing, and cellular targets as well as to facilitate rapid mutagenesis to define functional protein domains...
February 15, 2018: Plant Cell
Milorad Kojic, Mira Milisavljevic, William K Holloman
Cells maintain a small arsenal of resolving functions to process and eliminate complex DNA intermediates that result as a consequence of homologous recombination and distressed replication. Ordinarily the homologous recombination system serves as a high-fidelity mechanism to restore the integrity of a damaged genome, but in the absence of the appropriate resolving function it can turn DNA intermediates resulting from replication stress into pathological forms that are toxic to cells. Here we have investigated how the nucleases Mus81 and Gen1 and the helicase Blm contribute to survival after DNA damage or replication stress in Ustilago maydis cells with crippled yet homologous recombination-proficient forms of Brh2, the BRCA2 ortholog and primary Rad51 mediator...
February 2, 2018: DNA Repair
Christine L Kirkpatrick, Nicole C Parsley, Tessa E Bartges, Madeline E Cooke, Wilaysha S Evans, Lilian R Heil, Thomas J Smith, Leslie M Hicks
Fungal secondary metabolites represent a rich and largely untapped source for bioactive molecules, including peptides with substantial structural diversity and pharmacological potential. As methods proceed to take a deep dive into fungal genomes, complimentary methods to identify bioactive components are required to keep pace with the expanding fungal repertoire. We developed PepSAVI-MS to expedite the search for natural product bioactive peptides and herein demonstrate proof-of-principle applicability of the pipeline for the discovery of bioactive peptides from fungal secretomes via identification of the antifungal killer toxin KP4 from Ustilago maydis P4...
February 5, 2018: Journal of the American Society for Mass Spectrometry
Gabriel Schweizer, Karin Münch, Gertrud Mannhaupt, Jan Schirawski, Regine Kahmann, Julien Y Dutheil
Plants and fungi display a broad range of interactions in natural and agricultural ecosystems ranging from symbiosis to parasitism. These ecological interactions result in coevolution between genes belonging to different partners. A well-understood example is secreted fungal effector proteins and their host targets, which play an important role in pathogenic interactions. Biotrophic smut fungi (Basidiomycota) are well-suited to investigate the evolution of plant pathogens, because several reference genomes and genetic tools are available for these species...
February 1, 2018: Genome Biology and Evolution
Daniel Lanver, André N Müller, Petra Happel, Gabriel Schweizer, Fabian B Haas, Marek Franitza, Clément Pellegrin, Stefanie Reissmann, Janine Altmüller, Stefan A Rensing, Regine Kahmann
The corn smut fungus Ustilago maydis is a model organism for elucidating host colonization strategies of biotrophic fungi. Here we performed an in depth transcriptional profiling of the entire plant-associated development of U. maydis wild-type strains. In our analysis we focused on fungal metabolism, nutritional strategies, secreted effectors and regulatory networks. Secreted proteins were enriched in three distinct expression modules corresponding to stages on the plant surface, establishment of biotrophy and induction of tumors...
January 25, 2018: Plant Cell
Pei-Hua Chen, Jui-Yu Chou
Microbes have evolved ways of interference competition to gain advantage over their ecological competitors. The use of secreted antagonistic compounds by yeast cells is one of the prominent examples. Although this killer behavior has been thoroughly studied in laboratory yeast strains, our knowledge of the antagonistic specificity of killer effects in nature remains limited. In this study, yeast strains were collected from various niches and screened for antagonistic activity against one toxin-sensitive strain of Saccharomyces cerevisiae and three pathogenic fungi...
March 30, 2017: Polish Journal of Microbiology
Alexandra Matei, Corinna Ernst, Markus Günl, Björn Thiele, Janine Altmüller, Virginia Walbot, Björn Usadel, Gunther Doehlemann
The biotrophic fungus Ustilago maydis causes smut disease on maize (Zea mays), which is characterized by immense plant tumours. To establish disease and reprogram organ primordia to tumours, U. maydis deploys effector proteins in an organ-specific manner. However, the cellular contribution to leaf tumours remains unknown. We investigated leaf tumour formation at the tissue- and cell type-specific levels. Cytology and metabolite analysis were deployed to understand the cellular basis for tumourigenesis. Laser-capture microdissection was performed to gain a cell type-specific transcriptome of U...
March 2018: New Phytologist
Meilin Zhao, Xinyao Lu, Hong Zong, Jinyang Li, Bin Zhuge
Itaconic acid, 2-methylidenebutanedioic acid, is a precursor of polymers, chemicals, and fuels. Many fungi can synthesize itaconic acid; Aspergillus terreus and Ustilago maydis produce up to 85 and 53 g l-1 , respectively. Other organisms, including Aspergillus niger and yeasts, have been engineered to produce itaconic acid. However, the titer of itaconic acid is low compared with the analogous major fermentation product, citric acid, for which the yield is > 200 g l-1 . Here, we review two types of pathway for itaconic acid biosynthesis as well as recent advances by metabolic engineering strategies and process optimization to enhance itaconic acid productivity in native producers and heterologous hosts...
March 2018: Biotechnology Letters
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