Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization
Identifieur interne : 002576 ( Main/Exploration ); précédent : 002575; suivant : 002577Sensitive and Specific Digoxigenin‐labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot‐blot Hybridization
Auteurs : L. Barbarossa [Italie] ; V. Savino [Italie]Source :
- 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.
Url:
DOI: 10.1111/j.1439-0434.2006.01102.x
Affiliations:
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Barbarossa</name><affiliation wicri:level="1"><country wicri:rule="url">Italie</country><wicri:regionArea>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</wicri:regionArea><wicri:noRegion>70126 Bari</wicri:noRegion></affiliation></author><author><name sortKey="Savino, V" sort="Savino, V" uniqKey="Savino V" first="V." last="Savino">V. Savino</name><affiliation wicri:level="1"><country wicri:rule="url">Italie</country><wicri:regionArea>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</wicri:regionArea><wicri:noRegion>70126 Bari</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Phytopathology</title><idno type="ISSN">0931-1785</idno><idno type="eISSN">1439-0434</idno><imprint><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2006-06">2006-06</date><biblScope unit="volume">154</biblScope><biblScope unit="issue">6</biblScope><biblScope unit="page" from="329">329</biblScope><biblScope unit="page" to="335">335</biblScope></imprint><idno type="ISSN">0931-1785</idno></series><idno type="istex">BB7DE7B99A54861466D00F8A915527DFF3AFD48B</idno><idno type="DOI">10.1111/j.1439-0434.2006.01102.x</idno><idno type="ArticleID">JPH1102</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0931-1785</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Citrus tristeza virus</term><term>detection</term><term>dot‐blot hybridization</term><term>riboprobes</term><term>total RNA</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml: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.</div></front></TEI><affiliations><list><country><li>Italie</li></country></list><tree><country name="Italie"><noRegion><name sortKey="Barbarossa, L" sort="Barbarossa, L" uniqKey="Barbarossa L" first="L." last="Barbarossa">L. Barbarossa</name></noRegion><name sortKey="Savino, V" sort="Savino, V" uniqKey="Savino V" first="V." last="Savino">V. Savino</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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