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Sina Krombach, Stefanie Reissmann, Saskia Kreibich, Florian Bochen, Regine Kahmann
The peroxisomal sterol carrier protein 2 (Scp2) of the biotrophic maize pathogen Ustilago maydis was detected in apoplastic fluid, suggesting that it might function as a secreted effector protein. Here we analyze the role of the scp2 gene during plant colonization. We used reverse genetics approaches to delete the scp2 gene, determined stress sensitivity and fatty acid utilization of mutants, demonstrated secretion of Scp2, used quantitative reverse transcription polymerase chain reaction for expression analysis and expressed GFP-Scp2 fusion proteins for protein localization...
June 13, 2018: New Phytologist
Jinny A Paul, R Margaret Wallen, Chen Zhao, Tieliu Shi, Michael H Perlin
The dimorphic switch from budding to filamentous growth is an essential morphogenetic transition many fungi utilize to cause disease in the host. Although different environmental signals can induce filamentous growth, the developmental programs associated with transmitting these different signals may differ. Here, we explore the relationship between filamentation and expression levels of ammonium transporters (AMTs) that also sense low ammonium for Ustilago maydis, the pathogen of maize. Overexpression of the high affinity ammonium transporter, Ump2, under normally non-inducing conditions, results in filamentous growth...
July 2018: Fungal Biology
Mira Milisavljevic, Jelena Petkovic, Jelena Samardzic, Milorad Kojic
After heavy exposure of Ustilago maydis cells to clastogens, a great increase in viability was observed if the treated cells were kept under starvation conditions. This restitution of viability is based on cell multiplication at the expense of the intracellular compounds freed from the damaged cells. Analysis of the effect of the leaked material on the growth of undamaged cells revealed opposing biological activity, indicating that U. maydis must possess cellular mechanisms involved not only in reabsorption of the released compounds from external environment but also in contending with their treatment-induced toxicity...
2018: Frontiers in Microbiology
Sandra K Hartmann, Yvonne Stockdreher, Georg Wandrey, Hamed Hosseinpour Tehrani, Thiemo Zambanini, Andreas J Meyer, Jochen Büchs, Lars M Blank, Markus Schwarzländer, Nick Wierckx
Maintenance of metabolic redox homeostasis is essential to all life and is a key factor in many biotechnological processes. Changes in the redox state of NAD affect metabolic fluxes, mediate regulation and signal transduction, and thus determine growth and productivity. Here we establish an in vivo monitoring system for the dynamics of the cytosolic NADH/NAD+ ratio in the basidiomycete Ustilago maydis using the ratiometric fluorescent sensor protein Peredox-mCherry. Metabolic redox dynamics were determined in the cytosol of living cells with high time resolution under biotechnologically relevant conditions, i...
May 22, 2018: Biochimica et Biophysica Acta
Teeratas Kijpornyongpan, Stephen J Mondo, Kerrie Barry, Laura Sandor, Juna Lee, Anna Lipzen, Jasmyn Pangilinan, Kurt LaButti, Matthieu Hainaut, Bernard Henrissat, Igor V Grigoriev, Joseph W Spatafora, M Catherine Aime
Ustilaginomycotina is home to a broad array of fungi including important plant pathogens collectively called smut fungi. Smuts are biotrophs that produce characteristic perennating propagules called teliospores, one of which, Ustilago maydis, is a model genetic organism. Broad exploration of smut biology has been hampered by limited phylogenetic resolution of Ustilaginiomycotina as well as an overall lack of genomic data for members of this subphylum. In this study, we sequenced eight Ustilaginomycotina genomes from previously unrepresented lineages, deciphered ordinal-level phylogenetic relationships for the subphylum, and performed comparative analyses...
May 15, 2018: Molecular Biology and Evolution
Denise Seitner, Simon Uhse, Michelle Gallei, Armin Djamei
The biotrophic pathogen Ustilago maydis, the causative agent of corn smut disease, infects one of the most important crops worldwide - Zea mays. To successfully colonize its host, U. maydis secretes proteins, known as effectors, that suppress plant defense responses and facilitate the establishment of biotrophy. In this work, we describe the U. maydis effector protein Cce1. Cce1 is essential for virulence and is upregulated during infection. Through microscopic analysis and in vitro assays, we show that Cce1 is secreted from hyphae during filamentous growth of the fungus...
May 10, 2018: Molecular Plant Pathology
Lay-Sun Ma, Lei Wang, Christine Trippel, Artemio Mendoza-Mendoza, Steffen Ullmann, Marino Moretti, Alexander Carsten, Jörg Kahnt, Stefanie Reissmann, Bernd Zechmann, Gert Bange, Regine Kahmann
To cause disease in maize, the biotrophic fungus Ustilago maydis secretes a large arsenal of effector proteins. Here, we functionally characterize the repetitive effector Rsp3 (repetitive secreted protein 3), which shows length polymorphisms in field isolates and is highly expressed during biotrophic stages. Rsp3 is required for virulence and anthocyanin accumulation. During biotrophic growth, Rsp3 decorates the hyphal surface and interacts with at least two secreted maize DUF26-domain family proteins (designated AFP1 and AFP2)...
April 27, 2018: Nature Communications
Lucero Romero-Aguilar, Mónica Montero-Lomeli, Juan Pablo Pardo, Guadalupe Guerra-Sánchez
The article shows how to implement the LD index assay, which is a sensitive microplate assay to determine the accumulation of triacylglycerols (TAGs) in lipid droplets (LDs). LD index is obtained without lipid extraction. It allows measuring the LDs content in high-throughput experiments under different conditions such as growth in rich or nitrogen depleted media. Albeit the method was described for the first time to study the lipid droplet metabolism in Saccharomyces cerevisiae, it was successfully applied to the basidiomycete Ustilago maydis...
April 3, 2018: Journal of Visualized Experiments: JoVE
Juliana Benevenuto, Natalia S Teixeira-Silva, Eiko E Kuramae, Daniel Croll, Claudia B Monteiro-Vitorello
Host specialization is a key evolutionary process for the diversification and emergence of new pathogens. However, the molecular determinants of host range are poorly understood. Smut fungi are biotrophic pathogens that have distinct and narrow host ranges based on largely unknown genetic determinants. Hence, we aimed to expand comparative genomics analyses of smut fungi by including more species infecting different hosts and to define orphans and positively selected genes to gain further insights into the genetics basis of host specialization...
2018: Frontiers in Microbiology
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...
May 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 20, 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...
March 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...
May 1, 2018: Applied and Environmental Microbiology
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