Differentiation of species and strains among filamentous fungi by DNA fingerprinting

Meyer, Wieland; Koch, Anne; Niemann, Claudia; Beyermann, Birgit; Epplen, Jörg T.; Börner, Thomas

doi:10.1007/bf00336493

Cited by 89 publications

(43 citation statements)

References 15 publications

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“…Therefore, all of the strains that are to be compared must be processed with one stock solution of premixed reagents in one run of the thermocycler used and loaded onto one gel for maximum information output. Figures 1 through 4 show the results of RAPD analyses when we used primers M13 (GAGGGTGGCGGTTCT) and (GACA), (24) and primers ACGGTCTTGG (36) and TGC CGAGCTG (3). To calculate levels of DNA relatedness among species, bands were checked for reproducibility in parallel experiments.…”

Section: Resultsmentioning

confidence: 99%

“…Samples were subjected to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) as described by Laemmli (23) DNA extraction. Chromosomal DNA was extracted rapidly by a modified hexadecyltrimethylammonium bromide method (24). A 50-mg portion of wet matter was harvested from solid medium and suspended in 0.4 ml of hexadecyltrimethylammonium bromide buffer (2% [wt/vol] hexadecyltrimethylammonium bromide, 0.1 M Tris-HCl [pH 8.01, 1.2 M NaC1, 20 mM EDTA, 5 mM dithiothreitol) in a 1.5-ml Eppendorf cup.…”

Section: Methodsmentioning

confidence: 99%

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Molecular Characterization and Application of Random Amplified Polymorphic DNA Analysis of Mrakia and Sterigmatomyces Species

Messner¹,

Prillinger²,

Altmann³

et al. 1994

International Journal of Systematic Bacteriology

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The qualitative and quantitative monosaccharide spectra of purified yeast cell walls revealed that there are three phylogenetically distinct lineages of sterigma-forming basidiomycetous yeasts: (i) Kurtzmanomyces and Sterigmatomyces species, which contain high levels of mannose; (ii) Tilletiopsis species, which contain glucose, galactose, and small amounts of mannose; and (iii) Fellomyces, Kockovaella, Sterigmatosporidium, and Tsuchiyaea species, which appear to be closely related on the basis of their high levels of glucose and the presence of xylose. The yeast cell wall neutral sugars of Sporobolomyces antarcticus and Sterigmatomyces aphidis were similar to those of members of the genus Tilletiopsis. However, the possibility that these taxa are conspecific was eliminated by the results of a random amplified polymorphic DNA (RAPD) analysis. The conspecificity of Mrakia jiigida and Mrakia nivalis, the conspecificity of Mrakia gelida and Mrakia stokesii, and the conspecificity of Sterigmatomyces halophilus and Sterigmatomyces indicus were confirmed by RAPD analysis results. RAPD analysis was found to be a simple and highly sensitive method which can be used to differentiate species at the DNA level; it can replace nuclear DNA-nuclear DNA hybridization experiments for species identification, characterization, and delimitation.The genus Mrakia was proposed for Leucosporidium fngidum as a new genus of the Filobasidiaceae by Yamada and Komagata (51). The following four species that formerly had been placed in the genus Leucosporidium were transferred to the genus Mrakia: Mrakia Jizgida, Mrakia gelida, Mrakia nivalis, and Mrakia stokesii (7, 8, 51). A homothallic, apomictic life cycle, self-sporulating simple holobasidia bearing terminal, sessile basidiospores, fermentation of sugars, the presence of ubiquinone Q-8, production of extracellular amyloid compounds, and yeast cell walls containing xylose are the most distinctive features of Mrakia species (14,26,32,51). The fact that the genus Mrakia should be separated from phragmobasidial, smut-like Leucosporidium species (31) has been corroborated by the results of determinations of parts of the 18s and 26s rRNA sequences (6, 9,48) and ultrastructural septa1 pore data (37).The Mrakia species originally defined on the basis of their ability to ferment glucose and their ability to utilize nitrate or various carbon compounds (cellobiose, maltose, maltose, melibiose, raffinose) for growth (7, 8) turned out to be questionable taxa after investigations of the electrophoretic enzyme patterns (39, 52) and parts of the 18s and 26s rRNA sequences (6, 9, 48). The electrophoretic enzyme patterns (52) suggested that M. nivalis and M. stokesii are synonyms of M. fngida and M. gelida, respectively.The genus Sterigmatomyces was introduced by Fell (5) for nonfilamentous, yeast-like fungi that are characterized by a unique method of cell division. The yeast cells produce one or more sterigmata, each of which gives rise to a single conidium.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Molecular Characterization and Application of Random Amplified Polymorphic DNA Analysis of Mrakia and Sterigmatomyces Species

Messner¹,

Prillinger²,

Altmann³

et al. 1994

International Journal of Systematic Bacteriology

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“…Interestingly, isolates grouped together as having similar morphology also display similar patterns of pectic enzyme expression, as revealed by pectic zymograms (Hutton et al, 1994). The aim of the present work was to use microsatellite-primed PCR fingerprinting (Meyer et al, 1991) to assess the level of genetic diversity in mycorrhizal endophytes isolated from hair roots of Woollsia pungens (Cav.) F. Muell.…”

Section: mentioning

confidence: 97%

Molecular diversity of ericoid mycorrhizal endophytes isolated from Woollsia pungens

1998

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One hundred and sixty-eight sterile endophytic mycelia were isolated from roots of four Woollsia pungens (Cav.) F. Muell. (Epacridaceae) plants collected from a field site in New South Wales, Australia. All isolates formed typical ericoid mycorrhizal structures when inoculated onto roots of Vaccinium macrocarpon Ait. (Ericaceae). Microsatellite-primed PCR fingerprints generated using the primers (GTG) & and (GACA) % indicated that considerable genetic diversity exists within the endophyte population. It was estimated that a minimum of 43 genetically distinct mycelial genets were present in the root systems of the sampled W. pungens population, with most genets confined to individual plants. Two genets, however, were present within the root systems of two adjacent plants. While most genets were represented by less than eight isolates, three genets contained up to 41 isolates, suggesting that root system colonization by some endophytic mycelia might be extensive.

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“…neoformans serotype A (DUMC strain H99), which was also provided by J. R. Perfect, were isolated and separated by pulse field gel electrophoresis and blotted onto Hybond-N nylon membranes (Amersham, Arlington Heights, IL, USA), as previously described [27,281. The core sequence of wild-type phage M13 or the oligonucleotides (GTG), or (GACA), was then hybridized to the karyotype blots, as previously described [9].…”

Section: Karyotyping and Dna Hybridizationmentioning

confidence: 99%

Polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive DNA sequences: Strain variation in Cryptococcus neoformans

1995

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Minisatellites and simple repetitive DNA sequence motifs are used as conventional oligonucleotide probes in DNA-hybridization-based fingerprinting. The same oligonucleotides can be used as single primers in the polymerase chain reaction (PCR) to generate individual PCR fingerprints. In this study, the simple repetitive sequences, (CA)8, (CT)8, (CAC)5, (GTG)5, (GACA)4 and (GATA)4, and a minisatellite core sequence derived from the wild-type phage M13 (5' GAGGGTGGCGGTTCT 3') were used as specific, single primers to amplify hypervariable repetitive DNA sequences during PCR analysis. The potential applications of this techniques are demonstrated with clinical isolates of the human pathogenic yeast, Cryptococcus neoformans. PCR fingerprint patterns have remained stable after long-term in vitro passage ( > 2 1/2 years to date). Hybridization of the primers to blots of electrophorectically separated chromosomes demonstrated that the target sequences recognized by most of the primers are dispersed through the entire yeast genome. Sequence analysis of the cloned bands obtained by PCR fingerprinting indicated that if the same or extremely similar, inversely oriented tandem repeats are located close to each other, when only one repeat-specific primer is used in the PCR, the region between these repeats is amplified. PCR fingerprinting has a wide range of current and potential applications to fungi, such as clarifying taxonomic questions, facilitating epidemiological studies and improving the diagnosis of mycotic diseases.

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Differentiation of species and strains among filamentous fungi by DNA fingerprinting

Cited by 89 publications

References 15 publications

Molecular Characterization and Application of Random Amplified Polymorphic DNA Analysis of Mrakia and Sterigmatomyces Species

Molecular Characterization and Application of Random Amplified Polymorphic DNA Analysis of Mrakia and Sterigmatomyces Species

Molecular diversity of ericoid mycorrhizal endophytes isolated from Woollsia pungens

Polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive DNA sequences: Strain variation in Cryptococcus neoformans

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