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dc.contributor.authorMurcia, N.
dc.contributor.authorSerra, P.
dc.contributor.authorOlmos, Antonio
dc.contributor.authorDurán-Vila, Núria
dc.date.accessioned2017-06-01T10:09:33Z
dc.date.available2017-06-01T10:09:33Z
dc.date.issued2009
dc.identifier.citationMurcia, N., Serra, P., Olmos, A., Duran-Vila, N. (2009). A novel hybridization approach for detection of citrus viroids. Molecular and cellular probes, 23(2), 95-102.
dc.identifier.issn0890-8508
dc.identifier.urihttp://hdl.handle.net/20.500.11939/4193
dc.description.abstractCitrus plants are natural hosts of several viroid species all belonging to the family Pospiviroidae. Previous attempts to detect viroids from field-grown species and cultivars yielded erratic results unless analyses were performed using Etrog citron a secondary bio-amplification host. To overcome the use of Etrog citron a number of RT-PCR approaches have been proposed with different degrees of success. Here we report the suitability of an easy to handle northern hybridization protocol for viroid detection of samples collected from field-grown citrus species and cultivars. The protocol involves: (i) Nucleic acid preparations from bark tissue samples collected from field-grown trees regardless of the growing season and storage conditions; (ii) Separation in 5% PAGE or 1% agarose, blotting to membrane and. xing; (iii) Hybridization with viroid-specific DIG-labelled probes and detection with anti-DIG-alkaline phosphatase conjugate and autoradiography with the CSPD substrate. The method has been tested with viroid-infected trees of sweet orange, lemon, mandarin, grapefruit, sour orange, Swingle citrumello, Tahiti lime and Mexican lime. This novel hybridization approach is extremely sensitive, easy to handle and shortens the time needed for reliable viroid indexing tests. The suitability of PCR generated DIG-labelled probes and the sensitivity achieved when the samples are separated and blotted from non-denaturing gels are discussed. (C) 2009 Elsevier Ltd. All rights reserved.
dc.language.isoen
dc.titleA novel hybridization approach for detection of citrus viroids
dc.typearticle
dc.date.issuedFreeFormAPR 2009
dc.identifier.doi10.1016/j.mcp.2008.12.007
dc.journal.abbreviatedTitleMol.Cell.Probes
dc.journal.issueNumber2
dc.journal.titleMolecular and cellular probes
dc.journal.volumeNumber23
dc.page.final102
dc.page.initial95
dc.rights.accessRightsopenAccess
dc.source.typeImpreso


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