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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Versita
2006
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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. |
format | Online Article Text |
id | pubmed-6275730 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Versita |
record_format | MEDLINE/PubMed |
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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