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Fungal Genetics and Biology: FG & B

Dylan Kessler, Hyunkyu Sang, Amanda Bousquet, Jonathan P Hulvey, Dawlyn Garcia, Siyeon Rhee, Yoichiro Hoshino, Toshihiko Yamada, Geunhwa Jung
Sclerotinia homoeocarpa is the causal organism of dollar spot in turfgrasses and is a multinucleate fungus with a history of resistance to multiple fungicide classes. Heterokaryosis gives rise to the coexistence of genetically distinct nuclei within a cell, which contributes to genotypic and phenotypic plasticity in multinucleate fungi. We demonstrate that field isolates, resistant to either a demethylation inhibitor or methyl benzimidazole carbamate fungicide, can form heterokaryons with resistance to each fungicide and adaptability to serial combinations of different fungicide concentrations...
January 10, 2018: Fungal Genetics and Biology: FG & B
Christina S Nødvig, Jakob B Hoof, Martin E Kogle, Zofia D Jarczynska, Jan Lehmbeck, Dorte K Klitgaard, Uffe H Mortensen
CRISPR-Cas9 technologies are revolutionizing fungal gene editing. Here we show that survival of specific Cas9/sgRNA mediated DNA double strand breaks (DSBs) depends on the non-homologous end-joining, NHEJ, DNA repair pathway and we use this observation to develop a tool to assess protospacer efficiency in Aspergillus nidulans. Moreover, we show that in NHEJ deficient strains, highly efficient marker-free gene targeting can be performed. Indeed, we show that even single-stranded oligo nucleotides efficiently works as repair templates of specific Cas9/sgRNA induced DNA DSBs in A...
January 8, 2018: Fungal Genetics and Biology: FG & B
James W Dohn, Alexander W Grubbs, C Elizabeth Oakley, Berl R Oakley
Technical advances in Aspergillus nidulans enable relatively easy deletion of genomic sequences, insertion of sequences into the genome and alteration of genomic sequences. To extend the power of this system we wished to create strains with several selectable markers in a common genetic background to facilitate multiple, sequential transformations. We have developed an approach, using the recycling of the pyrG selectable marker, that has allowed us to create new deletions of the biA, pabaA, choA, and lysB genes...
January 5, 2018: Fungal Genetics and Biology: FG & B
Huifang Wang, Zhangjiang He, Linli Luo, Xin Zhao, Zhuoyue Lu, Tingying Luo, Min Li, Yongjun Zhang
The aldo-keto reductases (AKRs) belong to the NADP-dependent oxidoreductase superfamily, which play important roles in various physiological functions in prokaryotic and eukaryotic organisms. However, many AKR superfamily members remain uncharacterized. Here, a downstream target gene of the HOG1 MAPK pathways coding for an aldo-keto reductase, named Bbakr1, was characterized in the insect fungal pathogen, Beauveria bassiana. Bbakr1 expression increased in response to osmotic and salt stressors, and oxidative and heavy metal (chromium) stress...
January 3, 2018: Fungal Genetics and Biology: FG & B
María Victoria Aguilar Pontes, Aleksandrina Patyshakuliyeva, Harm Post, Edita Jurak, Kristiina Hildén, Maarten Altelaar, Albert Heck, Mirjam A Kabel, Ronald P de Vries, Miia R Mäkelä
The white button mushroom Agaricus bisporus is one of the most widely produced edible fungus with a great economical value. Its commercial cultivation process is often performed on wheat straw and animal manure based compost that mainly contains lignocellulosic material as a source of carbon and nutrients for the mushroom production. As a large portion of compost carbohydrates are left unused in the current mushroom cultivation process, the aim of this work was to study wild-type A. bisporus strains for their potential to convert the components that are poorly utilized by the commercial strain A15...
December 22, 2017: Fungal Genetics and Biology: FG & B
Norio Takeshita
Cells are dynamic systems, the state of which undergoes constant alteration that results in morphological changes and movement. Many dynamic cellular processes that appear continuous are driven by underlying mechanisms that oscillate with distinct periods. For example eukaryotic cells do not grow continuously, but rather by pulsed extension of the periphery. Stepwise cell extension at the hyphal tips of several filamentous fungi was discovered 20 years ago, but only a few molecular details of the mechanism have been clarified since then...
December 8, 2017: Fungal Genetics and Biology: FG & B
Lauren M Segal, Richard A Wilson
Fungal interactions with plants can involve specific morphogenetic developments to access host cells, the suppression of plant defenses, and the establishment of a feeding lifestyle that nourishes the colonizer often-but not always-at the expense of the host. Reactive oxygen species (ROS) metabolism is central to the infection process, and the stage-specific production and/ or neutralization of ROS is critical to the success of the colonization process. ROS metabolism during infection is dynamic-sometimes seemingly contradictory-and involves endogenous and exogenous sources...
December 7, 2017: Fungal Genetics and Biology: FG & B
Kalev Adamson, Martin S Mullett, Halvor Solheim, Irene Barnes, Michael M Müller, Jarkko Hantula, Martti Vuorinen, Audrius Kacergius, Svetlana Markovskaja, Dmitry L Musolin, Kateryna Davydenko, Nenad Keča, Karli Ligi, Rasa D Priedite, Hanna Millberg, Rein Drenkhan
Dothistroma septosporum, a notorious pine needle pathogen with an unknown historical geographic origin and poorly known distribution pathways, is nowadays found almost in all areas inhabited by pines (Pinus spp.). The main aim of this study was to determine the relationship between North European and East Asian populations. In total, 238 Eurasian D. septosporum isolates from 11 countries, including 211 isolates from northern Europe, 16 isolates from Russian Far East and 11 isolates from Bhutan were analysed using 11 species specific microsatellite and mating type markers...
December 6, 2017: Fungal Genetics and Biology: FG & B
Changzhong Yu, Shanshan Wei, Xiaorong Han, Hanhan Liu, Mengxiao Wang, Meiling Jiang, Min Guo, Jie Dou, Changlin Zhou, Lingman Ma
Cbf-14 (RLLRKFFRKLKKSV), a designed peptide derived from cathelicidin family AMP, has proven to be potent against drug-resistant bacteria. In the present study, we investigated the anti-cryptococcal activity of Cbf-14 in vitro and in a pulmonary infection mouse model. Sensitivity test indicated that Cbf-14 possessed effective antifungal activity against Cryptococcus neoformans with an MIC of 4-16 µg/ml, and killing experiments showed that fungicidal activity was achieved after only 4 h treatment with Cbf-14 at 4× MIC concentrations in vitro...
December 5, 2017: Fungal Genetics and Biology: FG & B
Kamila Rząd, Sławomir Milewski, Iwona Gabriel
Amino acids constitute the key sources of nitrogen for growth of Candida albicans. In order to survive inside the host in different and rapidly changing environments, this fungus must be able to adapt via its expression of genes for amino acid metabolism. We analysed the ARO8, ARO9, YER152C, and BNA3 genes with regards to their role in the nutritional flexibility of C. albicans. CaAro8p is undoubtedly the most versatile enzyme among the aminotransferases investigated. It is involved in the catabolism of histidine, lysine, and aromatic amino acids as well as in L-Lys, Phe and Tyr biosynthesis...
November 30, 2017: Fungal Genetics and Biology: FG & B
Shunsuke Masuo, Airi Komatsuzaki, Norio Takeshita, Eriko Ito, Okazoe Takaaki, Shengmin Zhou, Naoki Takaya
Glycogen is a homopolymer of glucose and a ubiquitous cellular-storage carbon. This study investigated which Aspergillus nidulans genes are involved in glycogen metabolism. Gene disruptants of predicted glycogen synthase (gsyA) and glycogenin (glgA) genes accumulated less cellular glycogen than the wild type strain, indicating that GsyA and GlgA synthesize glycogen similarly to other eukaryotes. Meanwhile, gene disruption of gphA encoding glycogen phosphorylase increased the amount of glycogen to a higher degree than wild type during the stationary phase that accompanies carbon-source limitation...
November 22, 2017: Fungal Genetics and Biology: FG & B
Lucía Mojardín, Montserrat Vega, Fernando Moreno, Hans-Peter Schmitz, Jürgen J Heinisch, Rosaura Rodicio
The NAD+-dependent glycerol 3-phosphate dehydrogenase (KlGpd1) is an important enzyme for maintenance of the cytosolic redox balance. In the milk yeast Kluyveromyces lactis The enzyme is localized in peroxisomes and in the cytosol, indicating its requirement for the oxidation of NADH in both compartments. Klgpd1 mutants grow more slowly on glucose than wild-type cells and do not grow on ethanol as a sole carbon source. We studied the molecular basis of the latter phenotype and found that Gpd1 is required for high expression of KlICL1 and KlMLS1 which encode the key enzymes of the glyoxylate pathway isocitrate lyase and malate synthase, respectively...
November 22, 2017: Fungal Genetics and Biology: FG & B
Hye-Seon Kim, Jung-Eun Kim, Hokyoung Son, Daniel Frailey, Robert Cirino, Yin-Won Lee, Randall Duncan, Kirk J Czymmek, Seogchan Kang
Similar to animals and plants, external stimuli cause dynamic spatial and temporal changes of cytoplasmic Ca2+ in fungi. Such changes are referred as the Ca2+ signature and control cellular responses by modulating the activity or location of diverse Ca2+-binding proteins (CBPs) and also indirectly affecting proteins that interact with CBPs. To understand the mechanism underpinning Ca2+ signaling, therefore, characterization of how Ca2+ moves to and from the cytoplasm to create Ca2+ signatures under different conditions is fundamental...
November 22, 2017: Fungal Genetics and Biology: FG & B
Kihyuck Choi, Stephen M Marek
Phoma medicaginis (syn. Ascochyta medicaginicola Qchen &L. Cai) causes spring black stem and leaf spot, an important disease of alfalfa and annual medics. P. medicaginis forms uninucleate conidia in melanized pycnidia and is genetically tractable using Agrobacterium mediated transformation (ATMT), resulting in random integration of T-DNA that occasionally generates pycnidial mutants. The T-DNA tagged mutant, P265 displayed smaller pycnidia and more aerial hyphae than the wild type. A single T-DNA disrupted a putative noncanonical poly(A) RNA polymerase gene, Pmncpap1, which in yeast interacts with ribonucleotide reductase (RNR)...
November 15, 2017: Fungal Genetics and Biology: FG & B
Natasha T Forester, Geoffrey A Lane, Melle Steringa, Iain L Lamont, Linda J Johnson
The symbiosis between Epichloë festucae and its host perennial ryegrass (Lolium perenne) is a model system for mutualistic interactions in which the fungal endophyte grows between plant shoot cells and acquires host nutrients to survive. E. festucae synthesises the siderophore epichloënin A (EA) via SidN, a non-ribosomal peptide synthetase (NRPS). EA is involved in the acquisition of iron, an essential micronutrient, as part of the process of maintaining a stable symbiotic interaction. Here, we mutated a different NRPS gene sidC and showed that it is required for production of a second siderophore ferricrocin (FC)...
November 15, 2017: Fungal Genetics and Biology: FG & B
Gero Steinberg, Nicholas J Harmer, Martin Schuster, Sreedhar Kilaru
In ascomycete fungi, hyphal cells are separated by perforate septa, which allow cell-to-cell communication. To protect against extensive wound-induced damage, septal pores are sealed by peroxisome-derived Woronin bodies (WBs). The mechanism underpinning WB movement is unknown, but cytoplasmic bulk flow may "flush" WBs into the pore. However, some studies suggest a controlled and active mechanism of WB movement. Indeed, in the wheat pathogen Zymoseptoria tritici cellular ATP prevents WBs from pore sealing in unwounded cells...
November 15, 2017: Fungal Genetics and Biology: FG & B
Pengyi Zhang, Hongbin Li, Jie Cheng, April Y Sun, Liqing Wang, Gordana Mircevska, Richard Calderone, Dongmei Li
We have previously established that mitochondrial Complex I (CI) mutants of Candida albicans display reduced oxygen consumption, decreased ATP production, and increased reactive oxidant species (ROS) during cell growth. Using the Seahorse XF96 analyzer, the energetic phenotypes of Electron Transport Chain (ETC) complex mutants are further characterized in the current study. The underlying regulation of energetic changes in these mutants is determined in glucose and non-glucose conditions when compared to wild type (WT) cells...
November 13, 2017: Fungal Genetics and Biology: FG & B
Elke-Martina Jung, Erika Kothe, Marjatta Raudaskoski
Basidiomycetes feature a prolonged dikaryotic life stage. A dispute over open versus closed mitosis could be solved using in vivo fluorescence videomicroscopy of histone 2B::EGFP and Lifeact labeled Schizophyllum commune. It revealed nuclei to condense to approximately one fifth in diameter during mitotic prophase. In addition, the specifics of clamp cell formation typical of many basidiomycetes included an actin network at the future site of nuclear division, which allowed for cessation of nuclear movement and re-localization of one nucleus towards the emerging clamp cell, while the other divided along the hyphal axis...
November 9, 2017: Fungal Genetics and Biology: FG & B
Hazal Boral, Banu Metin, Aylin Döğen, Seyedmojtaba Seyedmousavi, Macit Ilkit
The incidence of fungal diseases has been increasing since 1980, and is associated with excessive morbidity and mortality, particularly among immunosuppressed patients. Of the known 625 pathogenic fungal species, infections caused by the genera Aspergillus, Candida, Cryptococcus, and Trichophyton are responsible for more than 300 million estimated episodes of acute or chronic infections worldwide. In addition, a rather neglected group of opportunistic fungi known as black yeasts and their filamentous relatives cause a wide variety of recalcitrant infections in both immunocompetent and immunosuppressed hosts...
November 1, 2017: Fungal Genetics and Biology: FG & B
Mingyu Ding, Qili Zhu, Yusi Liang, Jing Li, Xinyue Fan, Xiaoyang Yu, Fang He, Houjuan Xu, Yuancun Liang, Jinfeng Yu
Phospholipase D (PLD) is an important phospholipid hydrolase that plays critical roles in various biological processes in eukaryotic cells. However, little is known about its functions in plant pathogenic fungi. In this study, we identified three FgPLD genes in Fusarium graminearum that are homologous to the Saccharomyces cerevisiae Spo14 gene. We constructed deletion mutants of all three FgPLD genes using homologous recombination. Deletion of FgPLD1 (Δpld1), but not FgPLD2 or FgPLD3, affected hyphal growth, conidiation, and perithecium formation...
October 25, 2017: Fungal Genetics and Biology: FG & B
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