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Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression

This paper presents a method of Agrobacterium-mediated transformation for two diploid breeding lines of potato, and gives a detailed analysis of reporter gene expression. In our lab, these lines were also used to obtain tetraploid somatic hybrids. We tested four newly prepared constructs based on th...

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Autores principales: Nadolska-Orczyk, Anna, Pietrusinska, Aleksandra, Binka-Wyrwa, Agnieszka, Kuc, Dominik, Orczyk, Wacław
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Versita 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275730/
https://www.ncbi.nlm.nih.gov/pubmed/17160584
http://dx.doi.org/10.2478/s11658-006-0064-9
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author Nadolska-Orczyk, Anna
Pietrusinska, Aleksandra
Binka-Wyrwa, Agnieszka
Kuc, Dominik
Orczyk, Wacław
author_facet Nadolska-Orczyk, Anna
Pietrusinska, Aleksandra
Binka-Wyrwa, Agnieszka
Kuc, Dominik
Orczyk, Wacław
author_sort Nadolska-Orczyk, Anna
collection PubMed
description This paper presents a method of Agrobacterium-mediated transformation for two diploid breeding lines of potato, and gives a detailed analysis of reporter gene expression. In our lab, these lines were also used to obtain tetraploid somatic hybrids. We tested four newly prepared constructs based on the pGreen vector system containing the selection gene nptII or bar under the 35S or nos promoter. All these vectors carried gus under 35S. We also tested the pDM805 vector, with the bar and gus genes respectively under the Ubi1 and Act1 promoters, which are strong for monocots. The selection efficiency (about 17%) was highest in the stem and leaf explants after transformation with pGreen where nptII was under 35S. About half of the selected plants were confirmed via PCR and Southern blot analysis to be transgenic and, depending on the combination, 0 to 100% showed GUS expression. GUS expression was strongest in multi-copy transgenic plants where gus was under Act1. The same potato lines carrying multi-copy bar under Ubi1 were also highly resistant to the herbicide Basta. The suggestion of using Agrobacterium-mediated transformation of diploid lines of potato as a model crop is discussed herein.
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spelling pubmed-62757302018-12-10 Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression Nadolska-Orczyk, Anna Pietrusinska, Aleksandra Binka-Wyrwa, Agnieszka Kuc, Dominik Orczyk, Wacław Cell Mol Biol Lett Article This paper presents a method of Agrobacterium-mediated transformation for two diploid breeding lines of potato, and gives a detailed analysis of reporter gene expression. In our lab, these lines were also used to obtain tetraploid somatic hybrids. We tested four newly prepared constructs based on the pGreen vector system containing the selection gene nptII or bar under the 35S or nos promoter. All these vectors carried gus under 35S. We also tested the pDM805 vector, with the bar and gus genes respectively under the Ubi1 and Act1 promoters, which are strong for monocots. The selection efficiency (about 17%) was highest in the stem and leaf explants after transformation with pGreen where nptII was under 35S. About half of the selected plants were confirmed via PCR and Southern blot analysis to be transgenic and, depending on the combination, 0 to 100% showed GUS expression. GUS expression was strongest in multi-copy transgenic plants where gus was under Act1. The same potato lines carrying multi-copy bar under Ubi1 were also highly resistant to the herbicide Basta. The suggestion of using Agrobacterium-mediated transformation of diploid lines of potato as a model crop is discussed herein. Versita 2006-12-12 /pmc/articles/PMC6275730/ /pubmed/17160584 http://dx.doi.org/10.2478/s11658-006-0064-9 Text en © University of Wrocław, Poland 2006
spellingShingle Article
Nadolska-Orczyk, Anna
Pietrusinska, Aleksandra
Binka-Wyrwa, Agnieszka
Kuc, Dominik
Orczyk, Wacław
Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title_full Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title_fullStr Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title_full_unstemmed Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title_short Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression
title_sort diploid potato (solanum tuberosum l.) as a model crop to study transgene expression
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275730/
https://www.ncbi.nlm.nih.gov/pubmed/17160584
http://dx.doi.org/10.2478/s11658-006-0064-9
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