Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization
Identifieur interne : 001665 ( Istex/Corpus ); précédent : 001664; suivant : 001666Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization
Auteurs : L. Barbarossa ; V. SavinoSource :
- Journal of Phytopathology [ 0931-1785 ] ; 2006-06.
English descriptors
Abstract
A non‐radioactive dot‐blot hybridization assay for the successful detection of Citrus tristeza virus (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)‐labelled minus‐sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection.
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DOI: 10.1111/j.1439-0434.2006.01102.x
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ISTEX:BB7DE7B99A54861466D00F8A915527DFF3AFD48BLe document en format XML
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Two digoxigenin (DIG)‐labelled minus‐sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection.</div></front></TEI><istex><corpusName>wiley</corpusName><author><json:item><name>L. Barbarossa</name><affiliations><json:string>Authors’ address: Dipartimento di Protezione delle Piante e Microbiologia Applicata, Università degli Studi and Istituto di Virologia Vegetale del CNR, Sezione di Bari, Via Amendola 165/A, 70126 Bari, Italy (correspondence to L. Barbarossa. E‐mail: l.barbarossa@ba.ivv.cnr.it)</json:string></affiliations></json:item><json:item><name>V. Savino</name><affiliations><json:string>Authors’ address: Dipartimento di Protezione delle Piante e Microbiologia Applicata, Università degli Studi and Istituto di Virologia Vegetale del CNR, Sezione di Bari, Via Amendola 165/A, 70126 Bari, Italy (correspondence to L. Barbarossa. 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Barbarossa. E‐mail: <email>l.barbarossa@ba.ivv.cnr.it</email>)</unparsedAffiliation></affiliation></affiliationGroup><keywordGroup xml:lang="en"><keyword xml:id="k1"><i>Citrus tristeza virus</i></keyword><keyword xml:id="k2">detection</keyword><keyword xml:id="k3">dot‐blot hybridization</keyword><keyword xml:id="k4">riboprobes</keyword><keyword xml:id="k5">total RNA</keyword></keywordGroup><abstractGroup><abstract type="main" xml:lang="en"><title type="main">Abstract</title><p>A non‐radioactive dot‐blot hybridization assay for the successful detection of <i>Citrus tristeza virus</i> (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)‐labelled minus‐sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>DIG‐labelled RNA Probes for Routine Detection of CTV</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization</title></titleInfo><name type="personal"><namePart type="given">L.</namePart><namePart type="family">Barbarossa</namePart><affiliation>Authors’ address: Dipartimento di Protezione delle Piante e Microbiologia Applicata, Università degli Studi and Istituto di Virologia Vegetale del CNR, Sezione di Bari, Via Amendola 165/A, 70126 Bari, Italy (correspondence to L. 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E‐mail: l.barbarossa@ba.ivv.cnr.it)</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">2006-06</dateIssued><edition>Received October 12, 2005; accepted February 1, 2006</edition><copyrightDate encoding="w3cdtf">2006</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">6</extent><extent unit="tables">2</extent></physicalDescription><abstract lang="en">A non‐radioactive dot‐blot hybridization assay for the successful detection of Citrus tristeza virus (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)‐labelled minus‐sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection.</abstract><subject lang="en"><genre>keywords</genre><topic>Citrus tristeza virus</topic><topic>detection</topic><topic>dot‐blot hybridization</topic><topic>riboprobes</topic><topic>total RNA</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Phytopathology</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0931-1785</identifier><identifier type="eISSN">1439-0434</identifier><identifier type="DOI">10.1111/(ISSN)1439-0434</identifier><identifier type="PublisherID">JPH</identifier><part><date>2006</date><detail type="volume"><caption>vol.</caption><number>154</number></detail><detail type="issue"><caption>no.</caption><number>6</number></detail><extent unit="pages"><start>329</start><end>335</end><total>7</total></extent></part></relatedItem><identifier type="istex">BB7DE7B99A54861466D00F8A915527DFF3AFD48B</identifier><identifier type="DOI">10.1111/j.1439-0434.2006.01102.x</identifier><identifier type="ArticleID">JPH1102</identifier><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>Blackwell Publishing Ltd</recordOrigin></recordInfo></mods></metadata><enrichments><json:item><type>multicat</type><uri>https://api.istex.fr/document/BB7DE7B99A54861466D00F8A915527DFF3AFD48B/enrichments/multicat</uri></json:item></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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