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Fungal Genetics and Biology: FG & B

Qiang Wang, Paul A Cobine, Jeffrey J Coleman
The Fusarium oxysporum species complex (FOSC) is an economically important group of pathogenic filamentous fungi that are able to infect both animals and plants. Reverse genetic techniques, including gene disruption/deletion methods, to study these fungi are available although limitations exist resulting in decreased efficiency. Herein we describe a gene editing system developed using a F. oxysporum-optimized Cas9 ribonucleoprotein (RNP) and protoplast transformation method. The Cas9 protein and sgRNA were assembled to form a stable RNP in vitro and this complex was transferred into fungal protoplasts for gene editing with PEG-mediated transformation...
May 12, 2018: Fungal Genetics and Biology: FG & B
Simon Gohlke, Daniela Heine, Hans-Peter Schmitz, Hans Merzendorfer
Chitin is mainly formed by the chitin synthase III complex (CSIII) in yeast cells. This complex is considered to be composed of the catalytic subunit Chs3 and the regulatory subunit Chs4, both of which are phosphoproteins and transported to the plasma membrane by different trafficking routes. During cytokinesis, Chs3 associates with Chs4 and other proteins at the septin ring, which results in an active CSIII complex. In this study, we focused on the role of Chs4 as a regulatory subunit of the CSIII complex...
May 12, 2018: Fungal Genetics and Biology: FG & B
Sachin Jain, Relebohile Sekonyela, Ben Knox, Jonathan M Palmer, Anna Huttenlocher, Mehdi Kabbage, Nancy P Keller
LaeA is a conserved global regulator of secondary metabolism and development in filamentous fungi. Examination of Aspergillus fumigatus transcriptome data of laeA deletion mutants have been fruitful in identifying genes and molecules contributing to the laeA mutant phenotype. One of the genes significantly down regulated in A. fumigatus ΔlaeA is metR, encoding a bZIP DNA binding protein required for sulfur and methionine metabolism in fungi. LaeA and MetR deletion mutants exhibit several similarities including down regulation of sulfur assimilation and methionine metabolism genes and ability to grow on the toxic sulfur analog, sodium selenate...
May 9, 2018: Fungal Genetics and Biology: FG & B
Tim J H Baltussen, Jordy P M Coolen, Jan Zoll, Paul E Verweij, Willem J G Melchers
Aspergillus fumigatus is a saprophytic fungus that extensively produces conidia. These microscopic asexually reproductive structures are small enough to reach the lungs. Germination of conidia followed by hyphal growth inside human lungs is a key step in the establishment of infection in immunocompromised patients. RNA-Seq was used to analyze the transcriptome of dormant and germinating A. fumigatus conidia. Construction of a gene co-expression network revealed four gene clusters (modules) correlated with a growth phase (dormant, isotropic growth, polarized growth)...
April 26, 2018: Fungal Genetics and Biology: FG & B
Johan van den Hoogen, Natalie Verbeek-de Kruif, Francine Govers
The oomycete Phytophthora infestans is a notorious plant pathogen with potato and tomato as its primary hosts. Previous research showed that the heterotrimeric G-protein subunits Gα and Gβ have a role in zoospore motility and virulence, and sporangial development, respectively. Here, we present analyses of the gene encoding a Gγ subunit in P. infestans, Pigpg1. The overall similarity of PiGPG1 with non-oomycete Gγ subunits is low, with only the most conserved amino acids maintained, but similarity with its homologs in other oomycetes is high...
April 25, 2018: Fungal Genetics and Biology: FG & B
Zhengyi Zhen, Xinjing Xing, Meihua Xie, Le Yang, Xuewei Yang, Yaqing Zheng, Yuanli Chen, Ni Ma, Qing Li, Ke-Qin Zhang, Jinkui Yang
Mitogen-activated protein (MAP) kinase Slt2 is a key player in the cell-wall integrity pathway of budding yeast. In this study, we functionally characterized Slt2 orthologs AoSlt2 and MhSlt2 from the nematode-trapping fungi Arthrobotrys oligospora and Monacrosporium haptotylum, respectively. We found that disruption of AoSlt2 and MhSlt2 led to reduced mycelial growth, increased sensitivity to environmental stresses such as sodium dodecyl sulfate, Congo red, and H2 O2 , and an inability to produce conidia and nematode-trapping structures...
April 24, 2018: Fungal Genetics and Biology: FG & B
Masashi Yukawa, Tomoaki Yamauchi, Naoaki Kurisawa, Shakil Ahmed, Ken-Ichi Kimura, Takashi Toda
Many human cancer cells contain more than two centrosomes, yet these cancer cells can form pseudo-bipolar spindles through the mechanism, called centrosome clustering, and survive, instead of committing lethal multipolar mitoses. Kinesin-14/HSET, a minus end-directed motor, plays a crucial role in centrosome clustering. Accordingly, HSET is deemed to be a promising chemotherapeutic target to selectively kill cancer cells. Recently, three HSET inhibitors (AZ82, CW069 and SR31527) have been reported, but their specificity and efficacy have not been evaluated rigorously...
April 20, 2018: Fungal Genetics and Biology: FG & B
Miguel Hernanz-Koers, Mónica Gandía, Sandra Garrigues, Paloma Manzanares, Lynne Yenush, Diego Orzaez, Jose F Marcos
Current challenges in the study and biotechnological exploitation of filamentous fungi are the optimization of DNA cloning and fungal genetic transformation beyond model fungi, the open exchange of ready-to-use and standardized genetic elements among the research community, and the availability of universal synthetic biology tools and rules. The GoldenBraid (GB) cloning framework is a Golden Gate-based DNA cloning system developed for plant synthetic biology through Agrobacterium tumefaciens-mediated genetic transformation (ATMT)...
April 19, 2018: Fungal Genetics and Biology: FG & B
Matthew D Lebar, Jeffrey W Cary, Rajtilak Majumdar, Carol H Carter-Wientjes, Brian M Mack, Qijian Wei, Valdet Uka, Sarah De Saeger, José Diana Di Mavungu
Aspergillus flavus can colonize important food staples and produce aflatoxins, a group of toxic and carcinogenic secondary metabolites. Previous in silico analysis of the A. flavus genome revealed 56 gene clusters predicted to be involved in the biosynthesis of secondary metabolites. A. flavus secondary metabolites produced during infection of maize seed are of particular interest, especially with respect to their roles in the biology of the fungus. A predicted nonribosomal peptide synthetase-like (NRPS-like) gene, designated asaC (AFLA_023020), present in the uncharacterized A...
April 16, 2018: Fungal Genetics and Biology: FG & B
Opemipo Esther Fasoyin, Bin Wang, Mengguang Qiu, Xiaoyun Han, Kuang-Ren Chung, Shihua Wang
Carbon catabolite repression (CCR) is a very important mechanism employed in the utilization of carbon as an energy source, required for the regulation of growth, development and secondary metabolite production in fungi. Despite the wide study of this mechanism in fungi, little is known about the major CCR gene creA in A. flavus. Hence, we report identification of A. flavus carbon catabolite repression gene creA, which is responsible for the repression of secondary carbon sources. Gene deletion and over-expression was employed to explicate the role of creA in the morphology, pathogenicity, and secondary metabolite production in A...
April 12, 2018: Fungal Genetics and Biology: FG & B
Ning Xie, Gwenaël Ruprich-Robert, Philippe Silar, Eric Herbert, Roselyne Ferrari, Florence Chapeland-Leclerc
The Podospora anserina genome contains a large family of 15 multicopper oxidases (MCOs), including three genes encoding a FET3-like protein, an ABR1-like protein and an ascorbate oxidase (AO)-like protein. FET3, ABR1 and AO1 are involved in global laccase-like activity since deletion of the relevant genes led to a decrease of activity when laccase substrate (ABTS) was used as substrate. However, contrary to the P. anserina MCO proteins previously characterized, none of these three MCOs seemed to be involved in lignocellulose degradation and in resistance to phenolic compounds and oxidative stress...
April 11, 2018: Fungal Genetics and Biology: FG & B
Pankaj Kumar Singh, Soham Ray, Shallu Thakur, Rajeev Rathour, Vinay Sharma, Tilak Raj Sharma
Rice and Magnaporthe oryzae constitutes an ideal pathosystem for studying host-pathogen interaction in cereals crops. There are two alternative hypotheses, viz. Arms race and Trench warfare, which explain the co-evolutionary dynamics of hosts and pathogens which are under continuous confrontation. Arms race proposes that both R- and Avr genes of host and pathogen, respectively, undergo positive selection. Alternatively, trench warfare suggests that either R- or Avr- gene in the pathosystem is under balanced selection intending to stabilize the genetic advantage gained over the opposition...
April 6, 2018: Fungal Genetics and Biology: FG & B
André L Gomes-Vieira, Jeremy G Wideman, Lisvane Paes-Vieira, Suely L Gomes, Thomas A Richards, José R Meyer-Fernandes
The model yeast Saccharomyces cerevisiae elicits a transcriptional response to phosphate (Pi ) depletion. To determine the origins of the phosphate response (PHO) system, we bioinformatically identified putative PHO components in the predicted proteomes of diverse fungi. Our results suggest that the PHO system is ancient; however, components have been expanded or lost in different fungal lineages. To show that a similar physiological response is present in deeply-diverging fungi we examined the transcriptional and physiological response of PHO genes to Pi depletion in the blastocladiomycete Blastocladiella emersonii...
April 5, 2018: Fungal Genetics and Biology: FG & B
Gudrun Gygli, Ronald P de Vries, Willem J H van Berkel
Vanillyl alcohol oxidase (VAO) is a fungal flavoenzyme that converts a wide range of para-substituted phenols. The products of these conversions, e.g. vanillin, coniferyl alcohol and chiral aryl alcohols, are of interest for several industries. VAO is the only known fungal member of the 4-phenol oxidising (4PO) subgroup of the VAO/PCMH flavoprotein family. While the enzyme has been biochemically characterised in great detail, little is known about its physiological role and distribution in fungi. We have identified and analysed novel, fungal candidate VAOs and found them to be mostly present in Pezizomycotina and Agaricomycotina...
April 4, 2018: Fungal Genetics and Biology: FG & B
Wen Zhu, Jiasui Zhan, Bruce A McDonald
We combined a common garden experimental design with digital image analysis to determine how melanization responds to temperature and fungicide stress in 126 strains of Rhynchosporium commune sampled from nine global field populations. We found that different temperatures and fungicide stress significantly affected the degree of melanization. The nine field populations showed similar patterns in response to the different temperatures. Significant correlations were found between the degree of melanization and the local environment, including mean annual temperature, latitude, and relative humidity, suggesting that melanization is a locally adaptive trait...
April 4, 2018: Fungal Genetics and Biology: FG & B
Linghuo Jiang, Dayong Xu, Ahsan Hameed, Tianshu Fang, Abu Bakr Ahmad Fazili, Faiza Asghar
Pmr1 is the Golgi/ER calcium pump, while Rch1 is a newly identified negative regulator of calcium influx in the plasma membrane of yeast cells. We show here that CaRch1 plays a dominant role over CaPmr1 in response of Candida albicans to SDS and tunicamycin stresses, while CaPmr1 has a major role in cell wall stress. Deletion of CaRCH1 increases the calcium/calcineurin signaling level in cells lacking CaPMR1. Calcineurin function is required for the role of CaRch1 in SDS stresses, while it is required for the function of CaPmr1 under all conditions examined...
April 2, 2018: Fungal Genetics and Biology: FG & B
Adriana M Rico-Ramírez, Robert W Roberson, Meritxell Riquelme
In Neurospora crassa hyphae the localization of all seven chitin synthases (CHSs) at the Spitzenkörper (Spk) and at developing septa has been well analyzed. Hitherto, the mechanisms of CHSs traffic and sorting from synthesis to delivery sites remain largely unexplored. In Saccharomyces cerevisiae exit of Chs3p from the endoplasmic reticulum (ER) requires chaperone Chs7p. Here, we analyzed the role of CSE-7, N. crassa Chs7p orthologue in the biogenesis of CHS-4 (orthologue of Chs3p). In a N. crassa Δcse-7 mutant, CHS-4-GFP no longer accumulated at the Spk and septa...
March 27, 2018: Fungal Genetics and Biology: FG & B
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
Ioannis F Kalampokis, George C Kapetanakis, Konstantinos A Aliferis, George Diallinas
The development of fungicide-resistant fungal populations represents a major challenge for the agrochemical and agri-food sectors, which threatens food supply and security. The issue becomes complex for fungi that cause quantitative and qualitative losses due to mycotoxin biosynthesis. Nonetheless, currently, the molecular details underlying fungicide action and fungal resistance mechanisms are partially known. Here, we have investigated whether plasma membrane transporters contribute to specific fungicide uptake in the model fungus Aspergillus nidulans...
June 2018: Fungal Genetics and Biology: FG & B
Mikhail A Moldovan, Svetlana A Petrova, Mikhail S Gelfand
Riboswitches are conserved RNA structures located in non-coding regions of mRNA and able to bind small molecules (e.g. metabolites) changing conformation upon binding. This feature enables them to function as regulators of gene expression. The thiamin pyrophosphate (TPP) riboswitch is the only type of riboswitches found not only in bacteria, but also in eukaryotes - in plants, green algae, protists, and fungi. Two main mechanisms of fungal TPP riboswitch action, involving alternative splicing, have been established so far...
May 2018: Fungal Genetics and Biology: FG & B
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