Detection of Chalara fraxinea in common ash (Fraxinus excelsior) using real time PCR
Identifieur interne : 000F90 ( Istex/Corpus ); précédent : 000F89; suivant : 000F91Detection of Chalara fraxinea in common ash (Fraxinus excelsior) using real time PCR
Auteurs : A. Chandelier ; F. André ; F. LaurentSource :
- Forest Pathology [ 1437-4781 ] ; 2010-04.
Abstract
A real time PCR assay was developed for the detection of Chalara fraxinea in common ash. PCR primers and Taqman probes, based on the internal transcribed spacer region of the multi‐copy gene rDNA, were tested for specificity and sensitivity. The primers amplified an 81 bp fragment for C. fraxinea but did not amplify DNA from other Chalara species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. This molecular method could be useful for routine analysis of this emergent pathogen and for epidemiological studies.
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DOI: 10.1111/j.1439-0329.2009.00610.x
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PCR primers and Taqman probes, based on the internal transcribed spacer region of the multi‐copy gene rDNA, were tested for specificity and sensitivity. The primers amplified an 81 bp fragment for C. fraxinea but did not amplify DNA from other Chalara species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. This molecular method could be useful for routine analysis of this emergent pathogen and for epidemiological studies.</div></front></TEI><istex><corpusName>wiley</corpusName><author><json:item><name>A. Chandelier</name><affiliations><json:string>E‐mail: chandelier@cra.wallonie.be (for correspondence)</json:string></affiliations></json:item><json:item><name>F. 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The primers amplified an 81 bp fragment for C. fraxinea but did not amplify DNA from other Chalara species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. 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PCR primers and Taqman probes, based on the internal transcribed spacer region of the multi‐copy gene rDNA, were tested for specificity and sensitivity. The primers amplified an 81 bp fragment for C. fraxinea but did not amplify DNA from other Chalara species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. This molecular method could be useful for routine analysis of this emergent pathogen and for epidemiological studies.</p></abstract></profileDesc><revisionDesc><change when="2010-04">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/AFFDDD24F60A79E3FEEAEF42F782E479627A3923/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Wiley, elements deleted: body"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="yes"</istex:xmlDeclaration><istex:document><component version="2.0" type="serialArticle" xml:lang="en"><header><publicationMeta level="product"><publisherInfo><publisherName>Blackwell Publishing Ltd</publisherName><publisherLoc>Oxford, UK</publisherLoc></publisherInfo><doi origin="wiley" registered="yes">10.1111/(ISSN)1439-0329</doi><issn type="print">1437-4781</issn><issn type="electronic">1439-0329</issn><idGroup><id type="product" value="EFP"></id><id type="publisherDivision" value="ST"></id></idGroup><titleGroup><title type="main" sort="FOREST PATHOLOGY">Forest Pathology</title></titleGroup></publicationMeta><publicationMeta level="part" position="04002"><doi origin="wiley">10.1111/efp.2010.40.issue-2</doi><numberingGroup><numbering type="journalVolume" number="40">40</numbering><numbering type="journalIssue" number="2">2</numbering></numberingGroup><coverDate startDate="2010-04">April 2010</coverDate></publicationMeta><publicationMeta level="unit" type="article" position="2" status="forIssue"><doi origin="wiley">10.1111/j.1439-0329.2009.00610.x</doi><idGroup><id type="unit" value="EFP610"></id></idGroup><countGroup><count type="pageTotal" number="10"></count></countGroup><titleGroup><title type="tocHeading1">ORIGINAL ARTICLES</title></titleGroup><copyright>© 2009 Blackwell Verlag GmbH</copyright><eventGroup><event type="firstOnline" date="2009-06-05"></event><event type="publishedOnlineFinalForm" date="2010-04-13"></event><event type="xmlConverted" agent="Converter:BPG_TO_WML3G version:2.3.6 mode:FullText source:FullText result:FullText" date="2010-04-19"></event><event type="xmlConverted" agent="Converter:WILEY_ML3G_TO_WILEY_ML3GV2 version:4.0.1" date="2014-03-12"></event><event type="xmlConverted" agent="Converter:WML3G_To_WML3G version:4.1.7 mode:FullText,remove_FC" date="2014-10-17"></event></eventGroup><numberingGroup><numbering type="pageFirst" number="87">87</numbering><numbering type="pageLast" number="95">95</numbering></numberingGroup><linkGroup><link type="toTypesetVersion" href="file:EFP.EFP610.pdf"></link></linkGroup></publicationMeta><contentMeta><unparsedEditorialHistory>Received: 7.1.2009; accepted: 20.4.2009; editor: S. Woodward</unparsedEditorialHistory><countGroup><count type="figureTotal" number="3"></count><count type="tableTotal" number="3"></count></countGroup><titleGroup><title type="main">Detection of <i>Chalara fraxinea</i> in common ash (<i>Fraxinus excelsior</i>) using real time PCR</title><title type="shortAuthors"><i>A. Chandelier, F. André and F. Laurent</i></title><title type="short"><i>Detection of C. fraxinea in common ash</i></title></titleGroup><creators><creator creatorRole="author" xml:id="cr1" affiliationRef="#a2"><personName><givenNames>A.</givenNames><familyName>Chandelier</familyName></personName></creator><creator creatorRole="author" xml:id="cr2" affiliationRef="#a1"><personName><givenNames>F.</givenNames><familyName>André</familyName></personName></creator><creator creatorRole="author" xml:id="cr3" affiliationRef="#a1"><personName><givenNames>F.</givenNames><familyName>Laurent</familyName></personName></creator></creators><affiliationGroup><affiliation xml:id="a1" countryCode="BE"><unparsedAffiliation>Walloon Agricultural Research Centre, Department Biocontrol and Plant Genetic Resources, Rue de Liroux, 4; B‐5030 Gembloux, Belgium</unparsedAffiliation></affiliation><affiliation xml:id="a2"><unparsedAffiliation> E‐mail: <email>chandelier@cra.wallonie.be</email> (for correspondence)</unparsedAffiliation></affiliation></affiliationGroup><abstractGroup><abstract type="main" xml:lang="en"><title type="main">Summary</title><p>A real time PCR assay was developed for the detection of <i>Chalara fraxinea</i> in common ash. PCR primers and Taqman probes, based on the internal transcribed spacer region of the multi‐copy gene rDNA, were tested for specificity and sensitivity. The primers amplified an 81 bp fragment for <i>C. fraxinea</i> but did not amplify DNA from other <i>Chalara</i> species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. This molecular method could be useful for routine analysis of this emergent pathogen and for epidemiological studies.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Detection of Chalara fraxinea in common ash (Fraxinus excelsior) using real time PCR</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>Detection of C. fraxinea in common ash</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Detection of Chalara fraxinea in common ash (Fraxinus excelsior) using real time PCR</title></titleInfo><name type="personal"><namePart type="given">A.</namePart><namePart type="family">Chandelier</namePart><affiliation>E‐mail: chandelier@cra.wallonie.be (for correspondence)</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">F.</namePart><namePart type="family">André</namePart><affiliation>Walloon Agricultural Research Centre, Department Biocontrol and Plant Genetic Resources, Rue de Liroux, 4; B‐5030 Gembloux, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">F.</namePart><namePart type="family">Laurent</namePart><affiliation>Walloon Agricultural Research Centre, Department Biocontrol and Plant Genetic Resources, Rue de Liroux, 4; B‐5030 Gembloux, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article"></genre><originInfo><publisher>Blackwell Publishing Ltd</publisher><place><placeTerm type="text">Oxford, UK</placeTerm></place><dateIssued encoding="w3cdtf">2010-04</dateIssued><edition>Received: 7.1.2009; accepted: 20.4.2009; editor: S. Woodward</edition><copyrightDate encoding="w3cdtf">2010</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType><extent unit="figures">3</extent><extent unit="tables">3</extent></physicalDescription><abstract lang="en">A real time PCR assay was developed for the detection of Chalara fraxinea in common ash. PCR primers and Taqman probes, based on the internal transcribed spacer region of the multi‐copy gene rDNA, were tested for specificity and sensitivity. The primers amplified an 81 bp fragment for C. fraxinea but did not amplify DNA from other Chalara species or from other fungi isolated from ash, whether pathogenic or saprophytic. The limit of detection was 5 pg of genomic DNA per PCR. Moreover, naturally‐infected samples were correctly diagnosed. A procedure for DNA extraction from woody tissues using an electric drill yielded DNA of an appropriate quality for real time PCR. This molecular method could be useful for routine analysis of this emergent pathogen and for epidemiological studies.</abstract><relatedItem type="host"><titleInfo><title>Forest Pathology</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">1437-4781</identifier><identifier type="eISSN">1439-0329</identifier><identifier type="DOI">10.1111/(ISSN)1439-0329</identifier><identifier type="PublisherID">EFP</identifier><part><date>2010</date><detail type="volume"><caption>vol.</caption><number>40</number></detail><detail type="issue"><caption>no.</caption><number>2</number></detail><extent unit="pages"><start>87</start><end>95</end><total>10</total></extent></part></relatedItem><identifier type="istex">AFFDDD24F60A79E3FEEAEF42F782E479627A3923</identifier><identifier type="DOI">10.1111/j.1439-0329.2009.00610.x</identifier><identifier type="ArticleID">EFP610</identifier><accessCondition type="use and reproduction" contentType="copyright">© 2009 Blackwell Verlag GmbH</accessCondition><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>Blackwell Publishing Ltd</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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