MycorrhizaeV1, Main, Curation, bibRecord, 003871

Combining nested PCR and restriction digest of the internal transcribed spacer region to characterize arbuscular mycorrhizal fungi on roots from the field.

Identifieur interne : 003871 ( Main/Curation ); précédent : 003870; suivant : 003872

Combining nested PCR and restriction digest of the internal transcribed spacer region to characterize arbuscular mycorrhizal fungi on roots from the field.

Auteurs : Carsten Renker [Allemagne] ; Jochen Heinrichs ; Michael Kaldorf ; François Buscot

Source :

Mycorrhiza [ 0940-6360 ] ; 2003.

RBID : pubmed:12938031

Descripteurs français

English descriptors

KwdEn :
- Ascomycota (genetics), Base Sequence (MeSH), Basidiomycota (genetics), Biodiversity (MeSH), Chytridiomycota (genetics), DNA, Fungal (genetics), DNA, Ribosomal Spacer (genetics), Deoxyribonucleases, Type II Site-Specific (MeSH), Fungi (genetics), Molecular Sequence Data (MeSH), Mycorrhizae (genetics), Phylogeny (MeSH), Plant Roots (microbiology), Polymerase Chain Reaction (MeSH), Sequence Alignment (MeSH).
MESH :
- chemical , genetics : DNA, Fungal, DNA, Ribosomal Spacer.
- genetics : Ascomycota, Basidiomycota, Chytridiomycota, Fungi, Mycorrhizae.
- microbiology : Plant Roots.
- Base Sequence, Biodiversity, Deoxyribonucleases, Type II Site-Specific, Molecular Sequence Data, Phylogeny, Polymerase Chain Reaction, Sequence Alignment.

Abstract

Identification of arbuscular mycorrhizal fungi (AMF) on roots is almost impossible with morphological methods and, due to the presence of contaminating fungi, it is also difficult with molecular biological techniques. To allow broad investigation of the population structure of AMF in the field, we have established a new method to selectively amplify the internal transcribed spacer (ITS) region of most AMF with a unique primer set. Based on available sequences of the rDNA, one primer pair specific for AMF and a few other fungal groups was designed and combined in a nested PCR with the already established primer pair ITS5/ITS4. Amplification from contaminating organisms was reduced by an AluI restriction after the first reaction of the nested PCR. The method was assessed at five different field sites representing different types of habitats. Members of all major groups within the Glomeromycota (except Archaeosporaceae) were detected at the different sites. Gigasporaceae also proved detectable with the method based on cultivated strains.

DOI: 10.1007/s00572-002-0214-5
PubMed: 12938031

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Le document en format XML

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<country xml:lang="fr">Allemagne</country>
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<author><name sortKey="Heinrichs, Jochen" sort="Heinrichs, Jochen" uniqKey="Heinrichs J" first="Jochen" last="Heinrichs">Jochen Heinrichs</name>
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<author><name sortKey="Kaldorf, Michael" sort="Kaldorf, Michael" uniqKey="Kaldorf M" first="Michael" last="Kaldorf">Michael Kaldorf</name>
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<term>Biodiversity (MeSH)</term>
<term>Chytridiomycota (genetics)</term>
<term>DNA, Fungal (genetics)</term>
<term>DNA, Ribosomal Spacer (genetics)</term>
<term>Deoxyribonucleases, Type II Site-Specific (MeSH)</term>
<term>Fungi (genetics)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Mycorrhizae (genetics)</term>
<term>Phylogeny (MeSH)</term>
<term>Plant Roots (microbiology)</term>
<term>Polymerase Chain Reaction (MeSH)</term>
<term>Sequence Alignment (MeSH)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN fongique (génétique)</term>
<term>Alignement de séquences (MeSH)</term>
<term>Ascomycota (génétique)</term>
<term>Basidiomycota (génétique)</term>
<term>Biodiversité (MeSH)</term>
<term>Champignons (génétique)</term>
<term>Chytridiomycota (génétique)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Espaceur de l'ADN ribosomique (génétique)</term>
<term>Mycorhizes (génétique)</term>
<term>Phylogenèse (MeSH)</term>
<term>Racines de plante (microbiologie)</term>
<term>Réaction de polymérisation en chaîne (MeSH)</term>
<term>Séquence nucléotidique (MeSH)</term>
<term>Type II site-specific deoxyribonuclease (MeSH)</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Fungal</term>
<term>DNA, Ribosomal Spacer</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Ascomycota</term>
<term>Basidiomycota</term>
<term>Chytridiomycota</term>
<term>Fungi</term>
<term>Mycorrhizae</term>
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<term>Basidiomycota</term>
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<term>Chytridiomycota</term>
<term>Espaceur de l'ADN ribosomique</term>
<term>Mycorhizes</term>
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</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
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<term>Deoxyribonucleases, Type II Site-Specific</term>
<term>Molecular Sequence Data</term>
<term>Phylogeny</term>
<term>Polymerase Chain Reaction</term>
<term>Sequence Alignment</term>
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<term>Biodiversité</term>
<term>Données de séquences moléculaires</term>
<term>Phylogenèse</term>
<term>Réaction de polymérisation en chaîne</term>
<term>Séquence nucléotidique</term>
<term>Type II site-specific deoxyribonuclease</term>
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<front><div type="abstract" xml:lang="en">Identification of arbuscular mycorrhizal fungi (AMF) on roots is almost impossible with morphological methods and, due to the presence of contaminating fungi, it is also difficult with molecular biological techniques. To allow broad investigation of the population structure of AMF in the field, we have established a new method to selectively amplify the internal transcribed spacer (ITS) region of most AMF with a unique primer set. Based on available sequences of the rDNA, one primer pair specific for AMF and a few other fungal groups was designed and combined in a nested PCR with the already established primer pair ITS5/ITS4. Amplification from contaminating organisms was reduced by an AluI restriction after the first reaction of the nested PCR. The method was assessed at five different field sites representing different types of habitats. Members of all major groups within the Glomeromycota (except Archaeosporaceae) were detected at the different sites. Gigasporaceae also proved detectable with the method based on cultivated strains.</div>
</front>
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<Issue>4</Issue>
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<Title>Mycorrhiza</Title>
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<Abstract><AbstractText>Identification of arbuscular mycorrhizal fungi (AMF) on roots is almost impossible with morphological methods and, due to the presence of contaminating fungi, it is also difficult with molecular biological techniques. To allow broad investigation of the population structure of AMF in the field, we have established a new method to selectively amplify the internal transcribed spacer (ITS) region of most AMF with a unique primer set. Based on available sequences of the rDNA, one primer pair specific for AMF and a few other fungal groups was designed and combined in a nested PCR with the already established primer pair ITS5/ITS4. Amplification from contaminating organisms was reduced by an AluI restriction after the first reaction of the nested PCR. The method was assessed at five different field sites representing different types of habitats. Members of all major groups within the Glomeromycota (except Archaeosporaceae) were detected at the different sites. Gigasporaceae also proved detectable with the method based on cultivated strains.</AbstractText>
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Combining nested PCR and restriction digest of the internal transcribed spacer region to characterize arbuscular mycorrhizal fungi on roots from the field.

Combining nested PCR and restriction digest of the internal transcribed spacer region to characterize arbuscular mycorrhizal fungi on roots from the field.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki