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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 53 (2004), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: A gene encoding malate synthase, a key enzyme of the glyoxylate cycle, has been cloned and characterized in the necrotrophic wheat pathogen Stagonospora nodorum. Expression studies of Mls1 showed high levels of transcript in ungerminated spores whereas malate synthase enzyme activities were low. Expression studies in planta found that Mls1 transcript levels decreased ≈ 10-fold upon germination before slowly increasing throughout the remainder of the infection. To characterize Mls1 further, the gene was disrupted in S. nodorum by homologous recombination. In the absence of any supplied carbon source, the mls1 spores were unable to germinate and consequently the mutants were non-pathogenic. Germination and pathogenicity could be restored by the addition of either glucose or sucrose, implying that S. nodorum is reliant upon the catabolism of lipids for infection. Furthermore, analysis of lipid bodies in the mutant strain indicated that lipid mobilization and, consequently, peroxisomal β-oxidation of fatty acids is delayed or inhibited by the disruption of the glyoxylate cycle. This study has demonstrated for the first time in a fungal phytopathogen the requirement of malate synthase for pathogenicity, suggesting that gluconeogenesis is both dependent on the glyoxylate cycle and required for infection.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1546-1718
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] New diseases of humans, animals and plants emerge regularly. Enhanced virulence on a new host can be facilitated by the acquisition of novel virulence factors. Interspecific gene transfer is known to be a source of such virulence factors in bacterial pathogens (often manifested as pathogenicity ...
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0983
    Keywords: Protoplast fusion ; Plant pathogen ; Genetic analysis ; Fungi imperfecti
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of a pAN7-1-transformed isolated of Cladosporium fulvum race 4, harbouring a tandem duplication of the vector, and an untransformed race 5 isolate were fused, without selection for the presence of vector sequences. Fusion products were allowed to haploidize spontaneously. The inheritance of pAN7-1 sequences and the expression of the hph gene was studied in 85 progeny. A very high proportion (80%) of the progeny contained vector sequences and 70% of the progeny were resistant to hygromycin. Inactivation of the hph gene occurred in eight progeny. Rearrangement of vector sequences can account for the inactivation: there is no evidence for any RIP-like mechanism. A number of novel bands were observed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Cladosporium fulvum ; Ascomycete ; Phylogeny ; Ribosomal DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequence of part of the ribosomal DNA from races of the fungal tomato pathogen Cladosporium fulvum and other Cladosporium species have been determined. Comparisons of the internal transcribed spacer regions (ITS1 and ITS2) of several C. fulvum races showed complete sequence homology suggesting a recent evolutionary divergence. Comparisons of these nucleotide sequences in the ITS region with those of other Cladosporium species showed the close relationship within the Cladosporium genus. Using the nucleotide sequence of part of the 18s ribosomal subunit from these isolates and comparing them with sequences of some Ascomycetes, Basidiomycetes and Chytridiomycetes, obtained from GenBank, we infer the phylogeny of the Cladosporium species studied here. Our analysis shows that the Cladosporia form a monophyletic group which falls within the order Ascomycotina.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 24 (1993), S. 278-278 
    ISSN: 1432-0983
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Imperfect fungus ; RAPDs ; Genetic mapping ; Mitotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPD markers have been obtained in Cladosporium fulvum, in order to establish a genetic map based on mitotic recombination in this imperfect fungus. Segregation analysis has provided molecular evidence of a high degree of recombination during the parasexual cycle, and of the ploidy level of the parasexual progeny. A molecular study of 49 RAPDs obtained showed that, with only one exception, all RAPD markers studied represent repetitive DNA. This situation precludes the direct use of these markers to either initiate chromosome walking to a gene of interest or for the assignment of linkage groups to electrophoretically-separated chromosomes. A simple reamplification test has been applied which permitted the quick discrimination between single-copy and repetitive DNA species, without the need for Southern analysis. Additionally, evidence is shown for the presence of single-stranded DNA species in the amplification products, and that these may be present in addition to their double-stranded counterparts. The reamplification test can also identify these DNA species, avoiding misinterpretation of polymorphic bands.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1617-4623
    Keywords: Leptosphaeria maculans ; Brassica pathogen ; Hygromycin ; Phleomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Leptosphaeria maculans, a fungal pathogen of Brassica spp., was successfully transformed with the vector pAN8-1, encoding phleomycin resistance. Protoplasts of a vigorous Phleor transformant were then retransformed using the partially homologous vector, pAN7-1 which encodes hygromycin B resistance. Retransformation of this strain to hygromycin resistance occurred at frequencies that were consistently twofold higher than with the original recipient strain. Linearised pAN7-1 DNA transformed phleomycin-resistant protoplasts at higher frequencies still. All the transformants that were tested retained a phleomycin-resistant phenotype (20/20). Molecular analysis of five transformants generated with circular pAN7-1 DNA indicated that in four cases the pAN7-1 vector had integrated into pAN8-1 sequences. These results suggest that transformation frequencies in L. maculans are limited by the ability of vector DNA to integrate into the genome. Hence, construction of strains with target sites for integration may prove to be a generally useful method for improving transformation frequencies of poorly characterised filamentous fungi, particularly when using heterologous vectors. This would greatly facilitate the identification of genes by transfer of gene libraries and the standardisation of chromosomal location effects in studies of expression of nested promoter deletions.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1617-4623
    Keywords: Transposable elements ; Leaf mould ; Virus-like particles ; Reverse transcriptase ; Fulvia fulva
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A retrotransposon from the fungal tomato pathogen Cladosporium fulvum (syn. Fulvia fulva) has been isolated and characterised. It is 6968 by in length and bounded by identical long terminal repeats of 427 bp; 5 by target-site duplications were found. Putative first- and second-strand primer binding sites were identified. Three long open reading frames (ORFs) are predicted from the sequence. The first has homology to retroviral gag genes. The second includes sequences homologous to protease, reverse transcriptase, RNAse H and integrase, in that order. Sequence comparisons of the predicted ORFs indicate that this element is closely related to the gypsy class of LTR retrotransposons. Races of the pathogen exhibit polymorphisms in their complement of at least 25 copies of the sequence. Virus-like particles which co-sediment with reverse transcriptase activity were observed in homogenates of the fungus. This is the first report of an LTR retrotransposon in a filamentous fungus.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 249 (1995), S. 432-438 
    ISSN: 1617-4623
    Keywords: Transposon ; Aspergillus niger ; Filamentous fungus ; Nitrate reductase gene ; Tc1/mariner
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A transposable element has been isolated from the industrially important fungus Aspergillus niger (strain N402). The element was identified as an insertion sequence within the coding region of the nitrate reductase gene. It had inserted at a TA site and appeared to have duplicated the target site upon insertion. The isolated element was found to be 4798 by in length and contained 37-bp inverted, imperfect, terminal repeats (ITRs). The sequence of the central region of the element revealed an open reading frame (designated ORF1) which showed similarity, at the amino acid level, to the transposase of the Tc1/mariner class of DNA transposons. Another sequence within the central region of the element showed similarity to the 3′ coding and downstream untranslated region of the amyA gene of A. niger. Sequence homology and structural features indicate that this element, which has been named Ant1 (A. niger transposon 1), is related to the Tc1/mariner group of DNA transposons. Ant1 is apparently present as a single copy in strain N402 of A. niger.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1617-4623
    Keywords: Electrophoretic karyotype ; Chromosomes ; Deuteromycete phytopathogen ; Cladosporium fulvum ; Chromosome length polymorphisms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Methods are described for the electrophoretic separation of chromosome-sized DNA molecules from the fungal tomato pathogen Cladosporium fulvum (syn. Fulvia fulva). Using a hexagonal electrode array and switching times of 75 min at 45 V for 14 days, nine bands could be resolved. By comparison with co-electrophoresed Aspergillus nidulans chromosomal DNA (which was resolved into seven bands), the sizes of the C. fulvum bands are estimated to be between 1.9 Mb and 5.4 Mb. The two largest bands are believed to be doublets, giving a minimum genome size of 44 Mb. Cloned probes for the ribosomal DNA repeat, an anonymous single copy fragment and a newly discovered retrotransposon were hybridized to blots of the pulsed field gels, demonstrating the use of this technique for genomic mapping. Most strains of C. fulvum had an identical pattern of bands. Two strains exhibited two polymorphisms which could be due to a translocation.
    Type of Medium: Electronic Resource
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