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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 are 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...
January 30, 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...
January 4, 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...
January 3, 2018: Biotechnology Letters
Serge Fotso, Paul Graupner, Quanbo Xiong, Jeffrey R Gilbert, Don Hahn, Cruz Avila-Adame, George Davis, Kengo Sumiyoshi
Three novel cyclodepsipeptides, alveolarides A (1), B (2), and C (3), each possessing the rare 2,3-dihydroxy-4-methyltetradecanoic acid unit and a β-phenylalanine amino acid residue, along with the known peptide scopularide were isolated and identified from the culture broth of Microascus alveolaris strain PF1466. The pure compounds were evaluated for biological activity, and alveolaride A (1) provided strong in vitro activity against the plant pathogens Pyricularia oryzae, Zymoseptoria tritici, and Ustilago maydis...
December 28, 2017: Journal of Natural Products
Matthias Kretschmer, Scott Lambie, Daniel Croll, James W Kronstad
The fungal pathogen Ustilago maydis causes disease on maize by mating to establish an infectious filamentous cell type that invades the host and induces tumours. We previously found that β-oxidation mutants were defective in virulence and did not grow on acetate. Here we demonstrate that acetate inhibits filamentation during mating and in response to oleic acid. We therefore examined the influence of different carbon sources by comparing the transcriptomes of cells grown on acetate, oleic acid or glucose, with expression changes for the fungus during tumour formation in planta...
December 13, 2017: Molecular Microbiology
Dario Rafael Olicón-Hernández, Cristina Uribe-Alvarez, Salvador Uribe-Carvajal, Juan Pablo Pardo, Guadalupe Guerra-Sánchez
Chitosan is a stressing molecule that affects the cells walls and plasma membrane of fungi. For chitosan derivatives, the action mode is not clear. In this work, we used the yeast Ustilago maydis to study the effects of these molecules on the plasma membrane, focusing on physiologic and stress responses to chitosan (CH), oligochitosan (OCH), and glycol-chitosan (GCH). Yeasts were cultured with each of these molecules at 1 mg·mL-1 in minimal medium. To compare plasma membrane damage, cells were cultivated in isosmolar medium...
December 7, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Yafen Zhang, Qianchao Cao, Peng Hu, Haifeng Cui, Xiaoping Yu, Zihong Ye
BACKGROUND: Ustilago esculenta, a pathogenic basidiomycete fungus, infects Zizania latifolia to form edible galls named Jiaobai in China. The distinct growth conditions of U. esculenta induced Z. latifolia to form three different phenotypes, named male Jiaobai, grey Jiaobai and white Jiaobai. The aim of this study is to characterize the genetic and morphological differences that distinguish the two U. esculenta strains. RESULTS: In this study, sexually compatible haploid sporidia UeT14/UeT55 from grey Jiaobai (T strains) and UeMT10/UeMT46 from white Jiaobai (MT strains) were isolated...
December 6, 2017: BMC Microbiology
Thiemo Zambanini, Sandra K Hartmann, Lisa M Schmitz, Linda Büttner, Hamed Hosseinpour Tehrani, Elena Geiser, Melanie Beudels, Dominik Venc, Georg Wandrey, Jochen Büchs, Markus Schwarzländer, Lars M Blank, Nick Wierckx
Background: Ustilago maydis is known for its natural potential to produce a broad range of valuable chemicals, such as itaconate, from both industrial carbon waste streams and renewable biomass. Production of itaconate, and many other secondary metabolites, is induced by nitrogen limitation in U. maydis. The clustered genes responsible for itaconate production have recently been identified, enabling the development of new expression tools that are compatible with biotechnological processes...
2017: Fungal Biology and Biotechnology
Angelika Czedik-Eysenberg, Franziska Rabe, Heinz Ekker, Carmen Czepe, Armin Djamei
We have established the Ustilago bromivora-Brachypodium spp. interaction as a new model pathosystem for biotrophic fungal plant infections of the head smut type (Rabe et al., 2016). In this protocol, the methodology used for comparing gene expression between saprophytic and in planta growth of the fungus is described. The experimental and analytical pipeline, how next generation RNA sequencing (Illumina RNA-Seq) analysis can be used to obtain lists of genes significantly up or down regulated in planta in comparison to axenic culture is given...
August 5, 2017: Bio-protocol
Thiemo Zambanini, Hamed Hosseinpour Tehrani, Elena Geiser, Christiane K Sonntag, Joerg M Buescher, Guido Meurer, Nick Wierckx, Lars M Blank
Ustilago trichophora RK089 has been found recently as a good natural malic acid producer from glycerol. This strain has previously undergone adaptive laboratory evolution for enhanced substrate uptake rate resulting in the strain U. trichophora TZ1. Medium optimization and investigation of process parameters enabled titers and rates that are able to compete with those of organisms overexpressing major parts of the underlying metabolic pathways. Metabolic engineering can likely further increase the efficiency of malate production by this organism, provided that basic genetic tools and methods can be established for this rarely used and relatively obscure species...
June 2017: Metabolic Engineering Communications
Sabrina Zander, Kira Müntjes, Michael Feldbrügge
An essential feature of protein expression is the tight regulation of when and where a protein is translated from its cognate mRNA. This spatiotemporal expression is particularly important in guaranteeing the correct and efficient targeting of proteins to defined subcellular sites. In order to achieve local translation, mRNAs must be deposited at specific locations. A common mechanism is the active transport of mRNAs along the actin or microtubule cytoskeleton. To study such dynamic transport processes in vivo RNA live imaging is the method of choice...
2018: Methods in Molecular Biology
Zhi-Dan Wang, Ning Yan, Zheng-Hong Wang, Xiao-Huan Zhang, Jing-Ze Zhang, Hui-Min Xue, Li-Xia Wang, Qi Zhan, Ying-Ping Xu, De-Ping Guo
We report a transcriptome assembly and expression profiles from RNA-Seq data and identify genes responsible for culm gall formation in Zizania latifolia induced by Ustilago esculenta. The smut fungus Ustilago esculenta can induce culm gall in Zizania latifolia, which is used as a vegetable in Asian countries. However, the underlying molecular mechanism of culm gall formation is still unclear. To characterize the processes underlying this host-fungus association, we performed transcriptomic and expression profiling analyses of culms from Z...
October 26, 2017: Plant Molecular Biology
Eun Young Yu, Min Hsu, William K Holloman, Neal F Lue
Homologous recombination and repair factors are known to promote both telomere replication and recombination-based telomere extension. Herein we address the diverse contributions of several recombination/repair proteins to telomere maintenance in Ustilago maydis, a fungus that bears strong resemblance to mammals with respect to telomere regulation and recombination mechanisms. In telomerase-positive U. maydis, deletion of rad51 and blm separately caused shortened but stably maintained telomeres, whereas deletion of both engendered similar telomere loss, suggesting that the repair proteins help to resolve similar problems in telomere replication...
October 20, 2017: Molecular Microbiology
Yanyong Cao, Lifeng Wang, Liping Duan, Jingjing Li, Juan Ma, Shuna Xie, Lei Shi, Huiyong Li
The common smut of corn, caused by Ustilago maydis is a troublesome disease of maize. Early and accurate detection of U. maydis is essential for the disease management. In this study, primer set Pep-2 was selected for LAMP (loop-mediated isothermal amplification) from 12 sets of primers targeting three U. maydis effector genes See1, Pit2 and Pep1 according to primer screening. The optimal concentrations of Bst DNA polymerase and Mg(2+) as well as inner/outer primer ratio of the LAMP reaction system were screened by combining a single factor experiment and an orthogonal design arrangement...
October 17, 2017: Scientific Reports
Dierk Niessing, Ralf-Peter Jansen, Thomas Pohlmann, Michael Feldbrügge
Eukaryotic cells rely on the precise determination of when and where proteins are synthesized. Spatiotemporal expression is supported by localization of mRNAs to specific subcellular sites and their subsequent local translation. This holds true for somatic cells as well as for oocytes and embryos. Most commonly, mRNA localization is achieved by active transport of the molecules along the actin or microtubule cytoskeleton. Key factors are molecular motors, adaptors, and RNA-binding proteins that recognize defined sequences or structures in cargo mRNAs...
October 10, 2017: Wiley Interdisciplinary Reviews. RNA
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