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Agrobacterium-mediated transformation of grapefruit with the wild-type and mutant RNA-dependent RNA polymerase genes of Citrus tristeza virus

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dc.creator Cevik, Bayram
dc.creator Niblett, Charles L.
dc.creator Lee, Richard F.
dc.date 2011-12-31T22:00:00Z
dc.date.accessioned 2020-10-06T11:26:32Z
dc.date.available 2020-10-06T11:26:32Z
dc.identifier e104054d-822d-47c5-bae6-7733fec9b5ae
dc.identifier 10.3906/tar-1008-1192
dc.identifier https://avesis.sdu.edu.tr/publication/details/e104054d-822d-47c5-bae6-7733fec9b5ae/oai
dc.identifier.uri http://acikerisim.sdu.edu.tr/xmlui/handle/123456789/74266
dc.description Citrus paradisi Macfad. 'Duncan' was transformed with constructs coding for the wild-type and mutant RNA-dependent RNA polymerase (RdRp) of Citrus tristeza virus (CTV) for exploring replicase-mediated pathogen-derived resistance (RM-PDR). The RdRp gene was amplified from a CTV genome and used to generate the wild-type and 2 mutant RdRp constructs for plant transformation. One mutant had the key amino acids GDD changed to AAA (RdRp-mGDD), and the second mutant had a deletion encompassing the GDD domain (RdRp-Delta GDD). Etiolated epicotyl segments of Duncan grapefruit (Citrus paradisi Macfad. 'Duncan') were transformed with each of these constructs using the Agrobacterium-mediated transformation method. From 4540 transformed epicotyl segments, 1402 kanamycin-resistant shoots were regenerated. After testing for expression of green fluorescent protein (GFP) and beta-glucuronidase (GUS) reporter genes by fluorescence microscopy and histochemical staining, respectively, 146 GUS-positive plants were rooted and 97 surviving plants were established in soil in pots. A total of 70 plants were tested for the presence of the GUS gene and CTV RdRp transgenes by polymerase chain reaction (PCR). A total of 51 GUS and CTV transgene-positive transgenic plants (15 with RdRp, 21 with RdRp-mGDD, and 15 with RdRp-Delta GDD) were identified.
dc.language eng
dc.rights info:eu-repo/semantics/closedAccess
dc.title Agrobacterium-mediated transformation of grapefruit with the wild-type and mutant RNA-dependent RNA polymerase genes of Citrus tristeza virus
dc.type info:eu-repo/semantics/article


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