Fungal Genetics Reports最新文献

{"title":"Application of the nourseothricin acetyltransferase gene (nat1) as dominant marker for the transformation of filamentous fungi","authors":"U. Kück, Birgit Hoff, Lehrstuhl für Allgemeine","doi":"10.4148/1941-4765.1106","DOIUrl":"https://doi.org/10.4148/1941-4765.1106","url":null,"abstract":"Here, we report the construction of two transformation vectors, pD-NAT1 and pG-NAT1, carrying the nat1 gene encoding the nourseothricin acetyltransferase. The nat1 gene is expressed under the control of the Aspergillus nidulans trpC promoter and thus can be used as a dominant drug-resistance marker for the DNA-mediated transformation of filamentous fungi. The successful application of both vectors was demonstrated by transforming the homothallic ascomycete Sordaria macrospora as well as the β-lactam producer Acremonium chrysogenum. For both fungi and for both vectors, transformation frequencies were between 10 and 40 transformants per 10 µg of plasmid DNA.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"10 1","pages":"9-11"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74815144","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":"Podospora anserina AS6 gene encodes the cytosolic ribosomal protein of the E. coli S12 family","authors":"M. Déquard-Chablat, P. Silar","doi":"10.4148/1941-4765.1111","DOIUrl":"https://doi.org/10.4148/1941-4765.1111","url":null,"abstract":"The ribosomal proteins of the E. coli S4, S5 and S12 families that are part of the ribosome accuracy center control translation accuracy both in prokaryotes and eukaryotes. In Podospora anserina, genes coding for S4 and S5 have already been identified. Here, we identify the gene coding for the S12 protein homologue and show that it is identical to the genetically known AS6 gene. 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/vol53/iss1/8 26 Fungal Genetics Newsletter Podospora anserina AS6 gene encodes the cytosolic ribosomal protein of the E. coli S12 family Michelle Dequard-Chablat and Philippe Silar 1 1, 2 Institut de Génétique et Microbiologie, Bât. 400 Université de Paris 11, 91405 Orsay cedex, France. 1 UFR de Biochimie, Université de Paris 7 Denis Diderot, case 7006, 2 place Jussieu, 75005, Paris, France 2 To whom correspondence may be addressed: michelle.chablat@igmors.u-psud.fr or philippe.silar@igmors.u-psud.fr Fungal Genetics Newsletter 53: 26-29 The ribosomal proteins of the E. coli S4, S5 and S12 families that are part of the ribosome accuracy center control translation accuracy both in prokaryotes and eukaryotes. In Podospora anserina, genes coding for S4 and S5 have already been identified. Here, we identify the gene coding for the S12 protein homologue and show that it is identical to the genetically known AS6 gene. Podospora anserina has been used in intensive search of translation accuracy mutants (Coppin-Raynal et al. 1988). Several factors involved in the maintenance of accuracy have been identified in this organism including the tRNA suppressors su4-1 and su8-1 (Debuchy et al. 1985), as well as elongation factor eEF1A coded by AS4 (Silar et al. 1994), termination factors eRF1 and eRF3 coded by su1 and su2/AS2 respectively (Gagny et al. 1998), ribosomal proteins S12 coded by AS1 (Dequard-Chablat et al. 1994), S7 coded by su12 (Silar et al. 1997) and S1 coded by su3 (Silar et al. 2003). S12, S7 and S1 refer to the P. anserina numbering for ribosomal proteins (Dequard-Chablat et al. 1986) since the su12 and su3 genes code for the ribosomal proteins homologues of the E. coli S4 and S5, respectively. These two proteins are part of an accuracy center that has been conserved for more than two billion years in both prokaryotes and eukaryotes (Alksne et al. 1993). The center contains a third protein corresponding to the E. coli S12 protein, which remains to be identified in P. anserina. This protein is highly conserved and essential in all prokaryotes and eukaryotes investigated to date (Alksne et al. 1993). To identify the gene coding this protein in P. anserina, we took advantage of the availability of the complete genomic sequence of this fungus (available at http://podospora.igmors.u-psud.fr). We first located on the sequence map the gene encoding the P. anserina protein of the S1","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"16 1","pages":"26-29"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89877267","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":"Influence of Lithium ions on conidiophore size in Neurospora crassa","authors":"B. Aase, I. W. Jolma, P. Ruoff","doi":"10.4148/1941-4765.1113","DOIUrl":"https://doi.org/10.4148/1941-4765.1113","url":null,"abstract":"Lithium (Li) ions are known to affect Neurospora crassa’s growth speed and circadian clock period, while elevated temperatures abolish these influences. We wondered whether Li has also an effect on conidia size. We used cryo-SEM to investigate this question and report here the results of 1720 measurements showing that at 20°C the long and short conidial axes are significantly reduced at high Li concentrations (10-15 mM), while the ratio between the long and short axes remains approximately constant. An increased temperature (30°C) appears to abolish the Li effect on conidia size. 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/vol53/iss1/10 34 Fungal Genetics Newsletter Influence of Lithium ions on conidiophore size in Neurospora crassa Bodil Aase, Ingunn W. Jolma and Peter Ruoff Department of Mathematics and Natural Science, University of Stavanger, N-4036 Stavanger, Norway corresponding author. email: peter.ruoff@uis.no. # Fungal Genetics Newsletter 53:34-36 Lithium (Li) ions are known to affect Neurospora crassa’s growth speed and circadian clock period, while elevated temperatures abolish these influences. We wondered whether Li has also an effect on conidia size. We used cryo-SEM to investigate this question and report here the results of 1720 measurements showing that at 20°C the long and short conidial axes are significantly reduced at high Li concentrations (10-15 mM), while the ratio between the long and short axes remains approximately constant. An increased temperature (30°C) appears to abolish the Li effect on conidia size. Lithium (Li) has a profound influence on Neurospora crassa’s growth rate and circadian period (Engelmann 1987; Davis 2000; Dunlap and Loros 2004). Typically, at extracellular concentrations of 10 mM LiCl, the growth rate is significantly reduced and the circadian clock begins to get disrupted (Engelmann 1987; Lakin-Thomas 1993; Jolma et al. 2006). Interestingly, increased temperature can abolish the Li effect, possibly by an increased dissociation between Li and its assumed targets (Jolma et al. 2006). Because of the macroscopically distinct differences in conidiation when Neurospora is grown in the presence or absence of Li, we wondered whether there might be also a difference in microscopic conidiation, for example conidia size. In order to answer this question, we performed a study using a Zeiss Supra VP35 scanning electron microscope (SEM) with a Polaron cryo stage. The bd a strain (FGSC #1859) was grown on Petri dishes in LD (12h:12h) at 20°C or 30°C using Vogel’s medium as previously described (Jolma et al. 2006). Li was added to the medium as LiCl. Samples were taken from Petri dishes that showed approximately the same amount of total growth (=growth speed X growth time). Because of the dependence of the speed of this organism’s growth on temperature and LiCl","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"29 1","pages":"34-36"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78227122","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":"Non-circadian light inducible rhythm in Aspergillus nidulans.","authors":"S. Schoustra","doi":"10.4148/1941-4765.1110","DOIUrl":"https://doi.org/10.4148/1941-4765.1110","url":null,"abstract":"Aspergillus nidulans grown under standard laboratory conditions does not show circadian rhythmic growth. The presence of a circadian clock was demonstrated in A. nidulans at the level of gene expression (Greene et al. 2003), but a visually observable rhythm is lacking. We recently observed a remarkable rhythmic growth pattern in the formation of conidiospores and ascospores in a fludioxonil resistant mutant of A. nidulans (fldA1) grown in a dark incubator. This is reminiscent of a circadian rhythm. We found however that our observed rhythm is induced by light (leaking into the ‘dark’ incubator) and is not self-sustainable. In absolute darkness or constant light the rhythm is lost; therefore, we conclude that the rhythm is not a true intrinsic circadian rhythm.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"68 1","pages":"23-25"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82389201","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":"High throughput mating tests in Neurospora crassa","authors":"K. McCluskey, Rachel L. Yedlin, S. Walker","doi":"10.4148/1941-4765.1109","DOIUrl":"https://doi.org/10.4148/1941-4765.1109","url":null,"abstract":"Mating type tests in Neurospora crassa are an important way to characterize strains. Since most of the knock-out mutants developed as part of the functional genomics program (Colot et al., 2006) have little obvious phenotype we have undertaken to test the mating type of all of the knock-strains that are sent to the FGSC. Our original mating type test protocol is similar to that described by Smith (1962) and involves growing a lawn of the fluffy tester strains (4317 or 4347) on cornmeal agar (Difco) in 15 cm petri plates. This is also described in the online Neurospora protocol, \"How to use fluffy testers for determining mating type and for other applications\" (http://www.fgsc.net/Neurospora/NeurosporaProtocolGuide.htm). The strains to be tested are grown on Vogels minimal (Vogel, 1956) or appropriately supplemented medium (McCluskey, 2003) and small amounts of conidia are transferred to a spot on a grid. Using this technique, thirty to forty strains can be tested on two plates. While robust, this technique is labor intensive and because the plate is opened for each inoculation there is the possibility that occasional stray conidia could confound the results.","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"6 1","pages":"20-22"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80352944","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":"Sequences important for heterokaryon incompatibility function in MAT A-1 of Neurospora crassa","authors":"P. Shiu, N. Glass","doi":"10.4148/1941-4765.1108","DOIUrl":"https://doi.org/10.4148/1941-4765.1108","url":null,"abstract":"Strains of Neurospora crassa exist as two alternative mating type forms, A and a; differences in mating type are required for the initiation of the sexual cycle (Shiu and Glass, 2000). The mating-type (mat) locus also acts as a heterokaryon incompatibility (het) locus, such that hyphal fusion between A and a strains results in a heterokaryon that shows extremely inhibited growth, absence of conidiation, and hyphal compartmentation and death (Glass et al., 2000). The A and a mating type sequences occupy the same locus in A and a strains, but are highly dissimilar in sequence. The mat a-1 gene, which encodes a putative HMG (high mobility group) type of transcriptional regulator, provides all the functions for the a mating type, including mating, ascospore formation, and heterokaryon incompatibility (Chang and Staben, 1994). The mat A locus encodes three proteins. MAT A-2 and MAT A-3 are responsible for ascospore formation (Ferreira et al., 1998); MAT A-3 is a putative HMG type of transcriptional regulator. MAT A-1 is predicted to be a a-domain type of transcriptional regulator and is both necessary and sufficient to confer A mating specificity and trigger heterokaryon incompatibility with a strains (Glass et al., 1990). Mutations in an unlinked locus, tol, suppress mating-type incompatibility such that tol A and tol a strains are capable of forming a vigorous heterokaryon (Newmeyer, 1970; Shiu and Glass, 1999).","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"24 1","pages":"15-19"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82556570","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 simple way to make a dilution series","authors":"R. L. Metzenberg","doi":"10.4148/1941-4765.1114","DOIUrl":"https://doi.org/10.4148/1941-4765.1114","url":null,"abstract":"Often one needs to determine a suitable concentration of a previously untested nutrient or inhibitor to use in subsequent experiments. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This brief note is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol53/iss1/11","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"33 1","pages":"37"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82581840","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":"Glycosyl Hydrolase Genes and Enzymes of Neurospora crassa","authors":"A. Radford","doi":"10.4148/1941-4765.1107","DOIUrl":"https://doi.org/10.4148/1941-4765.1107","url":null,"abstract":"An analysis of the genome of Neurospora crassa has identified genes encoding 84 putative glycosyl hydrolases, representing 24 different families in Henrissat's classification. Functionally, they include enzymes for the degradation of all major polysaccharides (including cellulase, hemicellulase, chitinase and pectinase).There is evidence of high levels of gene amplification, despite the presence of RIP, as there are eight representatives of family 3 (beta-glucosidases and xylosidases), five of family 7 (endoand exo-glucanases), six of family 13 (amylases and maltase), nine of family 18 (chitinase), eight of family 47 (ER alpha-mannosidases), eleven of family 61 (endoglucanases) and seven of family 76 (alpha-mannanases). 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/vol53/iss1/4","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"3 1","pages":"12-14"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72825098","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":"The effect of repeated freeze-thaw cycles on cryopreserved Neurospora crassa samples.","authors":"K. McCluskey, A. Wiest, S. Walker","doi":"10.4148/1941-4765.1115","DOIUrl":"https://doi.org/10.4148/1941-4765.1115","url":null,"abstract":"To better characterize handling parameters for the arrayed mutants prepared for the Neurospora functional genomics program, we have put 7 day old conidia from strain FGSC 2489 through a series of cycles of freezing at -80 C in 25% glycerol and 3.5% reconstituted non-fat dry milk. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This brief note is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol53/iss1/12","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"84 1","pages":"37"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86853973","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 ras-1 bd Mauriceville strain for mapping mutations in Oak Ridge ras-1 bd strains","authors":"Kaitlyn Beasley, T. Lamb, W. Versaw, Deborah Bell-Pedersen","doi":"10.4148/1941-4765.1112","DOIUrl":"https://doi.org/10.4148/1941-4765.1112","url":null,"abstract":"We describe the construction of a Neurospora crassa Mauriceville strain carrying the ras-1bd mutation marked by the bacterial hygromycin resistance gene, hph (new FGSC # 10156). This strain is valuable for mapping mutations in Oak Ridge strains that carry the bd mutation. 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/vol53/iss1/9 30 Fungal Genetics Newsletter A ras-1 Mauriceville strain for mapping mutations in Oak Ridge ras-1 strains bd bd A. Kaitlyn Beasley*, Teresa M. Lamb*, Wayne K. Versaw and Deborah Bell-Pedersen* *Center for Biological Clocks Research and Department of Biology, Texas A&M University, College Station, TX 77843 Fungal Genetics Newsletter 53:30-33 We describe the construction of a Neurospora crassa Mauriceville strain carrying the ras-1 mutation marked by the bacterial bd hygromycin resistance gene, hph (new FGSC # 10156). This strain is valuable for mapping mutations in Oak Ridge strains that carry the bd mutation. The bd mutation has been a benefit to circadian rhythm research as it slows down wild type growth rates and sharpens the conidial banding pattern on racetubes (Sargent et al., 1966; Bell-Pedersen et al., 2005). Recent work (Belden et al., 2006) has shown that the bd mutation lies in the ras-1 gene. Mutations that affect circadian banding patterns are typically isolated in strains carrying the ras-1 allele. Mapping such mutations by the CAPS method (Jin et al., in press) requires that the mutant phenotype be followed in bd a cross to a wild type Mauriceville strain. Assuming non-linkage, only half of the progeny will be bd, and thus readily scorable. Insertion of the ras-1 allele into the Mauriceville parent renders all progeny useful for segregation analysis. Furthermore, bd marking ras-1 with the hygromycin resistance gene, hph, permits a simple drug resistance test to determine linkage to ras-1. bd The strategy for gene replacement was similar to published split marker gene deletion strategies (Catlett et al., 2003; Colot et al., 2006). However, rather than deleting the wild type gene, we replaced it with the bd allele and inserted the hph gene in the 3'UTR at position + 225 from the stop codon (Figure 1). Figure 1. Schematic diagram of ras-1 genetic manipulations. Regions of homology are indicated by vertical hatch marks; there are 763 bp of homology between the split hph fragments, 3223 bp of homology at the 5' end, and 2582 bp of homology at the 3' end. Pvu II and Xho I sites are marked to show the differences between the wild type (wt) genomic (g) locus and the proper ras1 -hph integrant. The dashed line indicates a gap where the hph gene is integrated and the dotted lines indicate flanking genomic bd DNA that remains unchanged. The replacement fragments were generated in two steps. The first step involved amplification of ras-1 genomic regions from the bd O","PeriodicalId":12490,"journal":{"name":"Fungal Genetics Reports","volume":"11 1","pages":"30-33"},"PeriodicalIF":0.0,"publicationDate":"2006-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78141681","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}