Specific and sensitive PCR-based detection of Septoria musiva, S. populicola and S. populi, the causes of leaf spot and stem canker on poplars.
Identifieur interne : 003F51 ( Main/Exploration ); précédent : 003F50; suivant : 003F52Specific and sensitive PCR-based detection of Septoria musiva, S. populicola and S. populi, the causes of leaf spot and stem canker on poplars.
Auteurs : Nicolas Feau [Canada] ; Jerry E. Weiland ; Glen R. Stanosz ; Louis BernierSource :
- Mycological research [ 0953-7562 ] ; 2005.
Descripteurs français
- KwdFr :
- Alignement de séquences (MeSH), Amorces ADN (MeSH), Ascomycota (classification), Ascomycota (génétique), Ascomycota (isolement et purification), Données de séquences moléculaires (MeSH), Espaceur de l'ADN ribosomique (MeSH), Feuilles de plante (microbiologie), Maladies des plantes (microbiologie), Populus (microbiologie), Réaction de polymérisation en chaîne (méthodes), Sensibilité et spécificité (MeSH), Spécificité d'espèce (MeSH), Tiges de plante (microbiologie).
- MESH :
- génétique : Ascomycota.
- isolement et purification : Ascomycota.
- microbiologie : Feuilles de plante, Maladies des plantes, Populus, Tiges de plante.
- méthodes : Réaction de polymérisation en chaîne.
- Alignement de séquences, Amorces ADN, Ascomycota, Données de séquences moléculaires, Espaceur de l'ADN ribosomique, Sensibilité et spécificité, Spécificité d'espèce.
English descriptors
- KwdEn :
- Ascomycota (classification), Ascomycota (genetics), Ascomycota (isolation & purification), DNA Primers (MeSH), DNA, Ribosomal Spacer (MeSH), Molecular Sequence Data (MeSH), Plant Diseases (microbiology), Plant Leaves (microbiology), Plant Stems (microbiology), Polymerase Chain Reaction (methods), Populus (microbiology), Sensitivity and Specificity (MeSH), Sequence Alignment (MeSH), Species Specificity (MeSH).
- MESH :
- chemical : DNA Primers, DNA, Ribosomal Spacer.
- classification : Ascomycota.
- genetics : Ascomycota.
- isolation & purification : Ascomycota.
- methods : Polymerase Chain Reaction.
- microbiology : Plant Diseases, Plant Leaves, Plant Stems, Populus.
- Molecular Sequence Data, Sensitivity and Specificity, Sequence Alignment, Species Specificity.
Abstract
The development of a PCR assay for the detection of the poplar pathogenic fungi Septoria musiva (teleomorph Mycosphaerella populorum), S. populicola (M. populicola) and S. populi (M. populi) is described. Three pairs of species-specific PCR primers were designed using interspecific polymorphisms in the internal transcribed spacer (ITS) of nuclear ribosomal RNA gene (rDNA) repeats. The specificity of the three primer pairs was successfully tested on a collection of 40 S. musiva, 39 S. populicola and six S. populi isolates. Using stringent PCR conditions, no cross-reaction was observed with any of the isolates tested. The specificity of the PCR assay was further confirmed with DNA extracted from 12 additional Septoria species and 17 other fungal species obtained from stems or leaves of poplars. Specific amplification of the fragments for S. musiva and S. populicola was sensitive relatively to the technique used, detecting as low as 1 pg template DNA, and 10 pg of DNA of the target species in a background of 1 ng of DNA of the other species. Moreover, using DNA purified directly from disrupted conidia, it was possible to detect with a probability of 90%, using one unique PCR assay, the DNA equivalent of 166 conidia per microl of S. musiva and 156 conidia per microl of S. populicola. The procedures developed in this work can thus be applied for rapid and accurate detection and identification of Septoria species from poplars.
DOI: 10.1017/s0953756205003242
PubMed: 16209307
Affiliations:
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Le document en format XML
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Three pairs of species-specific PCR primers were designed using interspecific polymorphisms in the internal transcribed spacer (ITS) of nuclear ribosomal RNA gene (rDNA) repeats. The specificity of the three primer pairs was successfully tested on a collection of 40 S. musiva, 39 S. populicola and six S. populi isolates. Using stringent PCR conditions, no cross-reaction was observed with any of the isolates tested. The specificity of the PCR assay was further confirmed with DNA extracted from 12 additional Septoria species and 17 other fungal species obtained from stems or leaves of poplars. Specific amplification of the fragments for S. musiva and S. populicola was sensitive relatively to the technique used, detecting as low as 1 pg template DNA, and 10 pg of DNA of the target species in a background of 1 ng of DNA of the other species. Moreover, using DNA purified directly from disrupted conidia, it was possible to detect with a probability of 90%, using one unique PCR assay, the DNA equivalent of 166 conidia per microl of S. musiva and 156 conidia per microl of S. populicola. The procedures developed in this work can thus be applied for rapid and accurate detection and identification of Septoria species from poplars.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16209307</PMID><DateCompleted><Year>2005</Year><Month>11</Month><Day>03</Day></DateCompleted><DateRevised><Year>2019</Year><Month>11</Month><Day>09</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0953-7562</ISSN><JournalIssue CitedMedium="Print"><Volume>109</Volume><Issue>Pt 9</Issue><PubDate><Year>2005</Year><Month>Sep</Month></PubDate></JournalIssue><Title>Mycological research</Title><ISOAbbreviation>Mycol Res</ISOAbbreviation></Journal><ArticleTitle>Specific and sensitive PCR-based detection of Septoria musiva, S. populicola and S. populi, the causes of leaf spot and stem canker on poplars.</ArticleTitle><Pagination><MedlinePgn>1015-28</MedlinePgn></Pagination><Abstract><AbstractText>The development of a PCR assay for the detection of the poplar pathogenic fungi Septoria musiva (teleomorph Mycosphaerella populorum), S. populicola (M. populicola) and S. populi (M. populi) is described. Three pairs of species-specific PCR primers were designed using interspecific polymorphisms in the internal transcribed spacer (ITS) of nuclear ribosomal RNA gene (rDNA) repeats. The specificity of the three primer pairs was successfully tested on a collection of 40 S. musiva, 39 S. populicola and six S. populi isolates. Using stringent PCR conditions, no cross-reaction was observed with any of the isolates tested. The specificity of the PCR assay was further confirmed with DNA extracted from 12 additional Septoria species and 17 other fungal species obtained from stems or leaves of poplars. Specific amplification of the fragments for S. musiva and S. populicola was sensitive relatively to the technique used, detecting as low as 1 pg template DNA, and 10 pg of DNA of the target species in a background of 1 ng of DNA of the other species. Moreover, using DNA purified directly from disrupted conidia, it was possible to detect with a probability of 90%, using one unique PCR assay, the DNA equivalent of 166 conidia per microl of S. musiva and 156 conidia per microl of S. populicola. 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IdType="pubmed">16209307</ArticleId><ArticleId IdType="doi">10.1017/s0953756205003242</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Canada</li></country><region><li>Québec</li></region><settlement><li>Québec (ville)</li></settlement><orgName><li>Université Laval</li></orgName></list><tree><noCountry><name sortKey="Bernier, Louis" sort="Bernier, Louis" uniqKey="Bernier L" first="Louis" last="Bernier">Louis Bernier</name><name sortKey="Stanosz, Glen R" sort="Stanosz, Glen R" uniqKey="Stanosz G" first="Glen R" last="Stanosz">Glen R. Stanosz</name><name sortKey="Weiland, Jerry E" sort="Weiland, Jerry E" uniqKey="Weiland J" first="Jerry E" last="Weiland">Jerry E. Weiland</name></noCountry><country name="Canada"><region name="Québec"><name sortKey="Feau, Nicolas" sort="Feau, Nicolas" uniqKey="Feau N" first="Nicolas" last="Feau">Nicolas Feau</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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