ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.
Identifieur interne : 003B33 ( Main/Corpus ); précédent : 003B32; suivant : 003B34ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.
Auteurs : M. Gardes ; T D BrunsSource :
- Molecular ecology [ 0962-1083 ] ; 1993.
English descriptors
- KwdEn :
- Ascomycota (genetics), Base Sequence (MeSH), Basidiomycota (genetics), DNA Primers (genetics), DNA, Fungal (genetics), DNA, Ribosomal (genetics), Ecosystem (MeSH), Gene Amplification (MeSH), Molecular Sequence Data (MeSH), Plants (genetics), Plants (microbiology), Polymerase Chain Reaction (MeSH), Repetitive Sequences, Nucleic Acid (MeSH), Species Specificity (MeSH), Symbiosis (genetics).
- MESH :
- chemical , genetics : DNA Primers, DNA, Fungal, DNA, Ribosomal.
- genetics : Ascomycota, Basidiomycota, Plants, Symbiosis.
- microbiology : Plants.
- Base Sequence, Ecosystem, Gene Amplification, Molecular Sequence Data, Polymerase Chain Reaction, Repetitive Sequences, Nucleic Acid, Species Specificity.
Abstract
We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.
DOI: 10.1111/j.1365-294x.1993.tb00005.x
PubMed: 8180733
Links to Exploration step
pubmed:8180733Le document en format XML
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<author><name sortKey="Gardes, M" sort="Gardes, M" uniqKey="Gardes M" first="M" last="Gardes">M. Gardes</name>
<affiliation><nlm:affiliation>Department of Plant Pathology, University of California, Berkeley 94720.</nlm:affiliation>
</affiliation>
</author>
<author><name sortKey="Bruns, T D" sort="Bruns, T D" uniqKey="Bruns T" first="T D" last="Bruns">T D Bruns</name>
</author>
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<publicationStmt><idno type="wicri:source">PubMed</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.</title>
<author><name sortKey="Gardes, M" sort="Gardes, M" uniqKey="Gardes M" first="M" last="Gardes">M. Gardes</name>
<affiliation><nlm:affiliation>Department of Plant Pathology, University of California, Berkeley 94720.</nlm:affiliation>
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<author><name sortKey="Bruns, T D" sort="Bruns, T D" uniqKey="Bruns T" first="T D" last="Bruns">T D Bruns</name>
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<series><title level="j">Molecular ecology</title>
<idno type="ISSN">0962-1083</idno>
<imprint><date when="1993" type="published">1993</date>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Ascomycota (genetics)</term>
<term>Base Sequence (MeSH)</term>
<term>Basidiomycota (genetics)</term>
<term>DNA Primers (genetics)</term>
<term>DNA, Fungal (genetics)</term>
<term>DNA, Ribosomal (genetics)</term>
<term>Ecosystem (MeSH)</term>
<term>Gene Amplification (MeSH)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Plants (genetics)</term>
<term>Plants (microbiology)</term>
<term>Polymerase Chain Reaction (MeSH)</term>
<term>Repetitive Sequences, Nucleic Acid (MeSH)</term>
<term>Species Specificity (MeSH)</term>
<term>Symbiosis (genetics)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Primers</term>
<term>DNA, Fungal</term>
<term>DNA, Ribosomal</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Ascomycota</term>
<term>Basidiomycota</term>
<term>Plants</term>
<term>Symbiosis</term>
</keywords>
<keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plants</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>Ecosystem</term>
<term>Gene Amplification</term>
<term>Molecular Sequence Data</term>
<term>Polymerase Chain Reaction</term>
<term>Repetitive Sequences, Nucleic Acid</term>
<term>Species Specificity</term>
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<front><div type="abstract" xml:lang="en">We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.</div>
</front>
</TEI>
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<DateCompleted><Year>1994</Year>
<Month>06</Month>
<Day>16</Day>
</DateCompleted>
<DateRevised><Year>2019</Year>
<Month>09</Month>
<Day>14</Day>
</DateRevised>
<Article PubModel="Print"><Journal><ISSN IssnType="Print">0962-1083</ISSN>
<JournalIssue CitedMedium="Print"><Volume>2</Volume>
<Issue>2</Issue>
<PubDate><Year>1993</Year>
<Month>Apr</Month>
</PubDate>
</JournalIssue>
<Title>Molecular ecology</Title>
<ISOAbbreviation>Mol Ecol</ISOAbbreviation>
</Journal>
<ArticleTitle>ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.</ArticleTitle>
<Pagination><MedlinePgn>113-8</MedlinePgn>
</Pagination>
<Abstract><AbstractText>We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.</AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Gardes</LastName>
<ForeName>M</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Department of Plant Pathology, University of California, Berkeley 94720.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Bruns</LastName>
<ForeName>T D</ForeName>
<Initials>TD</Initials>
</Author>
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<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D003160">Comparative Study</PublicationType>
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<MedlineJournalInfo><Country>England</Country>
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<MeshHeadingList><MeshHeading><DescriptorName UI="D001203" MajorTopicYN="N">Ascomycota</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D001487" MajorTopicYN="N">Basidiomycota</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
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<MeshHeading><DescriptorName UI="D017931" MajorTopicYN="N">DNA Primers</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D004271" MajorTopicYN="N">DNA, Fungal</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
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<MeshHeading><DescriptorName UI="D004275" MajorTopicYN="N">DNA, Ribosomal</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
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<MeshHeading><DescriptorName UI="D017753" MajorTopicYN="N">Ecosystem</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005784" MajorTopicYN="N">Gene Amplification</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D010944" MajorTopicYN="N">Plants</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000382" MajorTopicYN="N">microbiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D016133" MajorTopicYN="N">Polymerase Chain Reaction</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012091" MajorTopicYN="N">Repetitive Sequences, Nucleic Acid</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013045" MajorTopicYN="N">Species Specificity</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013559" MajorTopicYN="N">Symbiosis</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
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