Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes

Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recover,,, of transgenic plants showing gene silencing in woody species, Mexican lime [Citrus...

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Autores principales: Domínguez, Alfons, Fagoaga, Carmen, Navarro, Luis, Moreno, Pedro, Pena, Leandro
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/5146
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author Domínguez, Alfons
Fagoaga, Carmen
Navarro, Luis
Moreno, Pedro
Pena, Leandro
author_browse Domínguez, Alfons
Fagoaga, Carmen
Moreno, Pedro
Navarro, Luis
Pena, Leandro
author_facet Domínguez, Alfons
Fagoaga, Carmen
Navarro, Luis
Moreno, Pedro
Pena, Leandro
author_sort Domínguez, Alfons
collection ReDivia
description Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recover,,, of transgenic plants showing gene silencing in woody species, Mexican lime [Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under nonselective conditions had silenced transgenes. and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified, When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines. it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly. here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events.
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spelling ReDivia51462025-04-25T14:45:30Z Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes Domínguez, Alfons Fagoaga, Carmen Navarro, Luis Moreno, Pedro Pena, Leandro Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recover,,, of transgenic plants showing gene silencing in woody species, Mexican lime [Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under nonselective conditions had silenced transgenes. and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified, When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines. it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly. here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events. 2017-06-01T10:11:47Z 2017-06-01T10:11:47Z 2002 JUN 2002 article Dominguez, A., Fagoaga, C., Navarro, L., Moreno, P., Pena, L. (2002). Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes. Molecular Genetics and Genomics, 267(4), 544-556. 1617-4615 http://hdl.handle.net/20.500.11939/5146 10.1007/s00438-002-0688-z en openAccess Impreso
spellingShingle Domínguez, Alfons
Fagoaga, Carmen
Navarro, Luis
Moreno, Pedro
Pena, Leandro
Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title_full Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title_fullStr Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title_full_unstemmed Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title_short Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes
title_sort regeneration of transgenic citrus plants under non selective conditions results in high frequency recovery of plants with silenced transgenes
url http://hdl.handle.net/20.500.11939/5146
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