Fungal Genetics Reports最新文献

{"title":"Differential sensitivity of Ustilago maydis to fungal antibiotics on simple and complex media","authors":"A. Verma, T. Kapros, J. H. Waterborg","doi":"10.4148/1941-4765.1002","DOIUrl":"https://doi.org/10.4148/1941-4765.1002","url":null,"abstract":"We have observed that the basidiomycete Ustilago maydis can be partially or completely resistant to antibiotics when grown in defined growth media. In synthetic medium based on the fully defined mixture of simple organic compounds and salts U. maydis displays near wild-type growth at concentrations of hygromycin that effectively kill cells in complex nutrient media. The antibiotics geneticin, nourseothricin and phleomycin had similar effects. In contrast, the fungicide carboxin was equally effective in all growth media tested. Our observations could guide selection of growth media for genetic transformation of Ustilago and other fungi when sensitivity to common antibiotics is used as a selectable marker. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol62/iss1/3 8 Differential sensitivity of Ustilago maydis to fungal antibiotics on simple and complex media Anju Verma, Tamas Kapros, and Jakob H. Waterborg * 1 Division of Plant Sciences and Bond Life Sciences Center, University of Missouri-Columbia, Columbia, MO 65211, USA; 2 Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri-Kansas City, Kansas City, MO 64110, USA. * Corresponding author Fungal Genetics Reports 62:813 We have observed that the basidiomycete Ustilago maydis can be partially or completely resistant to antibiotics when grown in defined growth media. In synthetic medium based on the fully defined mixture of simple organic compounds and salts U. maydis displays near wild-type growth at concentrations of hygromycin that effectively kill cells in complex nutrient media. The antibiotics geneticin, nourseothricin and phleomycin had similar effects. In contrast, the fungicide carboxin was equally effective in all growth media tested. Our observations could guide selection of growth media for genetic transformation of Ustilago and other fungi when sensitivity to common antibiotics is used as a selectable marker. Introduction Like many other laboratories we have successfully used the fungicide carboxin as a selectable marker in gene transformation (Brachmann et al., 2004; Brachmann et al., 2001; Fernandez-Alvarez et al., 2009; Kojic and Holloman, 2000; Topp et al., 2002) to knock-out variant-specifically histone H3 isotype loci (Verma et al., 2011). After transformation, transformants were cultured under continued selection until stable, non-heterokaryon clones could be isolated (Verma et al., 2011). These cultures were maintained on synthetic minimal media consisting of yeast nitrogen base (YNB), a standard mixture of small organic compounds and salts, with glucose (SD). When attempting to use other standard antibiotic selectable markers such as hygromycin, geneticin, nourseothricin and phleomycin (Brachmann et al., 2004; Kamper, 2004; Kojic and Holloman, 2000), selection was not observed. Other l","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"1 1","pages":"8-13"},"PeriodicalIF":0.0,"publicationDate":"2016-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83009112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fungal Genetics Reports #62","authors":"M. Sachs","doi":"10.4148/1941-4765.1000","DOIUrl":"https://doi.org/10.4148/1941-4765.1000","url":null,"abstract":"","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"50 1","pages":"1"},"PeriodicalIF":0.0,"publicationDate":"2016-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88951272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A <i>mus-51</i> RIP allele for transformation of <i>Neurospora crassa</i>.","authors":"Zachary J Smith,&nbsp;Stacy Bedore,&nbsp;Stephanie Spingler,&nbsp;Thomas M Hammond","doi":"10.4148/1941-4765.1001","DOIUrl":"https://doi.org/10.4148/1941-4765.1001","url":null,"abstract":"<p><p>This report describes the construction and characterization of <i>mus-51^RIP70</i> , an allele for high-efficiency targeted integration of transgenes into the genome of the model eukaryote <i>Neurospora crassa</i>. Two of the <i>mus-51^RIP70</i> strains investigated in this work (RZS27.10 and RZS27.18) can be obtained from the Fungal Genetics Stock Center. The two deposited strains are, to our knowledge, genetically identical and neither one is preferred over the other for use in <i>Neurospora</i> research.</p>","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"62 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5515490/pdf/nihms879885.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35188965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of a new branching mutant of Neurospora intermedia from nature","authors":"A. Mukati, H. Vyas, A. Vyas","doi":"10.4148/1941-4765.1004","DOIUrl":"https://doi.org/10.4148/1941-4765.1004","url":null,"abstract":"Neurospora has been studied extensively to understand various phenomenon of fungal growth and morphogenesis. Fungal morphogenesis is a complex process which includes important aspects like polarized extension of the hyphal tip and hyphal branching. Observations at the tip have shown that growth vesicles arrive at the tip from proximal locations, and then seem to be distributed into the tip dome by the spitzenkorper. No specific mechanism has been defined for branching. Several theories have been proposed for branching that can be broadly divided into two groups. One group supports the mechanism involving origin of branch initiation factors at the hyphal tips (Bachewich and Heath, 1997; Kaminskyj and Heath, 1996; Riquelme and Bartnicki-Garcia, 2004; Watters and Griffiths, 2001). Other group suggests that the signals for branch initiation originate from within the colony or mycelial body (Prosser and Trinci, 1979; Trinci, 1974; Watters, 2006). Watters et al., (2000) proposed that the branching initiation is not completely controlled by the tip but, to some extent by factors occurring at previous branch point. Trinci (1974) proposed that mutation or factor which reduces the maximum rate of tip extension without disturbing the rate of vesicle production would results in an increase in the frequency of branch initiation without reducing the overall rate of hypha formation. Recently it has been found that the fungal growth rate show relationship with the frequency of hyphal branching (Watters et al., 2008, 2011). In this paper we describe a naturally occurring mutant of N. intermedia which shows defect in branching and the inheritance of this defect in the progeny.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"1 1","pages":"1-8"},"PeriodicalIF":0.0,"publicationDate":"2015-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77196450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the long-term storage of Cryphonectria parasitica","authors":"Joshua C. Springer, A. Baines, Matthew T. Chansler, A. Jarosz","doi":"10.4148/1941-4765.1007","DOIUrl":"https://doi.org/10.4148/1941-4765.1007","url":null,"abstract":"Isolates of the Chestnut blight pathogen, Cryphonectria parasitica, from six populations in Michigan, were stored in the late 1990s as agar plugs of mycelium in vials of sterile water held at room temperature. Approximately 29% of the fungal isolates were infected with mycoviruses at the time of storage. Each isolate was tested for revivification effectiveness by taking aliquots from vials filled with agar plugs of C. parasitica and sterile water and plating onto potato dextrose agar. Average revivification success was 70.5% across populations with a range of 33—84% within populations. In situations where vials had dried out during storage, success was low (4%), while success for vials that retained sterile water averaged 90%. Most importantly however, is the loss of mycoviruses from stored isolates; only 2 of 119 stored mycovirus infected isolates still contained mycoviruses after storage, suggesting that the double-stranded RNA mycoviruses are degraded during storage. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol60/iss1/2","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"12 1","pages":"11-15"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89916058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimized primers and other critical conditions for efficient fusion PCR to generate knockout vectors in filamentous fungi.","authors":"Nicholas Harrison, Brad Cavinder, J. Townsend, F. Trail","doi":"10.4148/1941-4765.1006","DOIUrl":"https://doi.org/10.4148/1941-4765.1006","url":null,"abstract":"Methods to streamline functional studies of large numbers of genes are essential to fully utilize the significant genomic resources now available for fungi. Fusion PCR is often used to join pieces of DNA together, particularly in the construction of DNA fragments for gene replacement in fungi. Here we present high-efficiency primers which reliably direct fusion and amplification to generate constructs for gene knockouts.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"33 1","pages":"1-10"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76233216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the DNA sequence of the putative ABC transporter NCU09975 in Neurospora crassa strains carrying acriflavin resistance markers.","authors":"A. Wiest, S. Koch, K. McCluskey","doi":"10.4148/1941-4765.1012","DOIUrl":"https://doi.org/10.4148/1941-4765.1012","url":null,"abstract":"Genomic DNA sequence was determined for the putative Neurospora crassa ABC transporter NCU09975 from several different classical mutant strains including several acriflavin resistant mutants. The sensitivity of these strains to acriflavin was tested. While the open reading frame NCU09975 has multiple polymorphisms in strains sequenced for other purposes, none of the acriflavin resistant classical mutants tested had polymorphisms in the NCU09975 coding region or in the 195 bases upstream of the translation start site. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol59/iss1/4 Fungal Genetics Reports 59, 2012. 26 Analysis of the DNA sequence of the putative ABC transporter NCU09975 in Neurospora crassa strains carrying acriflavin resistance markers. Aric E. Wiest, Sarah Koch and Kevin McCluskey. Fungal Genetics Stock Center, University of MissouriKansas City, Kansas City, MO 94110 USA Fungal Genet Reports 59: 26 29 Genomic DNA sequence was determined for the putative Neurospora crassa ABC transporter NCU09975 from several different classical mutant strains including several acriflavin resistant mutants. The sensitivity of these strains to acriflavin was tested. While the open reading frame NCU09975 has multiple polymorphisms in strains sequenced for other purposes, none of the acriflavin resistant classical mutants tested had polymorphisms in the NCU09975 coding region or in the 195 bases upstream of the translation start site. Introduction While years of mutagenesis and subsequent characterization in Neurospora crassa produced one of the most densely saturated genetic maps (Perkins et al. 2001), many of the classical mutants are not yet associated with open reading frames from the genome sequence (Galagan et al. 2003). Among these are acriflavin resistant mutants which have been assigned to seven different loci. A subset of these genes also confer resistance to multiple related drugs, including acridine orange, malachite green, and aminotriazole (Akiyama and Nakashima 1996). Moreover some acriflavin resistance genes are dominant while others are recessive. Because we wanted to find a dominant selectable marker for transformation of Neurospora and because the abc-3-like ORF NCU09975 had a large number of polymorphisms in its primary sequence among a group strains subject to whole genome sequence (McCluskey et al. 2011) we undertook to characterize this locus in strains carrying the acriflavin resistance markers acr-1 and Acr-3. We additionally validated the acriflavin sensitivity in the classical mutant strains carrying polymorphisms in NCU09975. Materials and methods Strains were cultured on Vogel's minimal medium (Vogel 1956) using standard practices (Davis and De Serres 1970) (Table 1). Table 1. Strains and their characteristics FGSC number Genotype Marker Location Acriflavin sensitivity 2489 m","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"164 1","pages":"26-29"},"PeriodicalIF":0.0,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73376144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient tools to target DNA to Podospora anserina","authors":"M. Déquard-Chablat, Tan-Trung Nguyen, V. Contamine, S. H. Denmat, F. Malagnac","doi":"10.4148/1941-4765.1011","DOIUrl":"https://doi.org/10.4148/1941-4765.1011","url":null,"abstract":"Because, in filamentous fungi, integration of transgenes proceeds mainly through nonhomologous recombination, transformation results in a random distribution of these DNA sequences. Moreover, they are often found as multiple copies clustered at a single locus. Therefore, for a given transgene, expression may be highly variable among independent transformants. To get rid of both multiple copies and position effects, due to the impact of the chromatin structure upon transgene expression, we constructed tools to specifically target single copies at two well-defined loci, Pa_2_3690 and Pa_4_5450 (Espagne et al., 2008). These genes have been chosen for the following reasons: (1) they have a relatively stable expression during the entire Podospora’s life cycle, (2) complete deletion of their ORF does not lead to an abnormal phenotype (Bidard et al., 2011), (3) they are located on two different chromosomes and (4) their second division segregation (SDS) frequency is 80% and 90%, respectively. Besides, a highly efficient homologous recombination ΔPaKu70 strain is available (El-Khoury et al., 2008), which makes gene replacement fairly easy. Taking advantage of this strain, we designed two plasmid tools to target genomic integration of any DNA fragment.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"305 1","pages":"21-25"},"PeriodicalIF":0.0,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79819811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phylogenic analysis of additional Neurospora crassa isolates","authors":"T. Radic, S. Gastaldello, J. Diegmann, T. Roenneberg","doi":"10.4148/1941-4765.1010","DOIUrl":"https://doi.org/10.4148/1941-4765.1010","url":null,"abstract":"The ascomycete Neurospora crassa is classical model organisms in biology. So far, a phylogenetic analysis based on genomic sequences of four non-functional nuclear loci has been reported for 44 natural isolates of N. crassa. Three subgroups (clades) with a distinct geographical distribution have been identified: clade A (Caribbean Basin and Ivory Coast), clade B (Europe, Ivory Coast and India), and clade C (India). Here, we report the results of a phylogenetic analysis of 16 additional isolates. Six of these were from the Caribbean Basin, eight from Europe and one from Pakistan and one from Thailand. The previously described clades and their geographical distribution were generally confirmed. All Caribbean isolates belonged to clade A and all European isolates belonged to clade B, with the exception of one isolate from Italy, which also belonged to clade A, suggesting a transport from the Caribbean Basin or the Ivory Coast to Europe. Interestingly, the isolates from Pakistan and Thailand were found in a separate group, basal to all other clades. Their phylogenetic classification is not yet clear as they might belong to N. crassa but as well to N. perkinsii, potentially representing yet undescribed phylogenetic groups or species of Neurospora, or hybrids. Authors Tanja Radic, Silvia Gastaldello, Julia Diegmann, and Till Roenneberg This regular paper is available in Fungal Genetics Reports: https://newprairiepress.org/fgr/vol59/iss1/2","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"68 1","pages":"13-20"},"PeriodicalIF":0.0,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75393348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Strategies for the molecular genetic manipulation and visualization of the human fungal pathogen Penicillium marneffei","authors":"Kylie J. Boyce, Hayley E. Bugeja, Harshini Weerasinghe, Michael Payne, Lena Schreider","doi":"10.4148/1941-4765.1009","DOIUrl":"https://doi.org/10.4148/1941-4765.1009","url":null,"abstract":"P. marneffei has been established as an experimentally amenable system to study morphogenesis and pathogenicity. This paper describes the development of a number of tools, including numerous selectable markers, to expand the ease with which it can be genetically manipulated. Combined with strains engineered for homologous recombination of exogenous DNA, these tools facilitate efficient molecular genetic studies.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"5 1","pages":"1-12"},"PeriodicalIF":0.0,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81874209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}