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Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia
In 2017, various orange coloured petunia on the market turned out to be genetically modified (GM) without an official authorization for commercialization. Sequence analysis suggested these undeclared plants most probably originated from a plant transformation experiment performed in the 1980s. For a...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359168/ https://www.ncbi.nlm.nih.gov/pubmed/32684656 http://dx.doi.org/10.1007/s11240-020-01871-w |
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author | Haselmair-Gosch, Christian Nitarska, Daria Walliser, Benjamin Flachowsky, Henryk Marinovic, Silvija Halbwirth, Heidi |
author_facet | Haselmair-Gosch, Christian Nitarska, Daria Walliser, Benjamin Flachowsky, Henryk Marinovic, Silvija Halbwirth, Heidi |
author_sort | Haselmair-Gosch, Christian |
collection | PubMed |
description | In 2017, various orange coloured petunia on the market turned out to be genetically modified (GM) without an official authorization for commercialization. Sequence analysis suggested these undeclared plants most probably originated from a plant transformation experiment performed in the 1980s. For a deeper understanding how GM petunia entered classical breeding programmes worldwide, and whether they originated from a single source or not, we undertook a molecular genetic characterization of the T-DNA integration sites in different GM petunia cultivars and breeding lines. By means of genome walking, we isolated different T-DNA sequences, which are located at the junctions between the T-DNA(s) and the petunia DNA. Based on the results obtained we conclude that there are at least two T-DNA copies of different lengths. This is supported by Southern blot analysis. For T-DNA1, the 3′-junction sequence was isolated, whereas the 5′-junction remained unclear. In contrast, for T-DNA2, the 5′-junction sequence was isolated, whereas the sequence isolated from the 3′-region consists only of T-DNA, but did not include the junction from the T-DNA to the petunia DNA. We developed primers for event-specific PCRs and screened a set of three orange GM petunia cultivars and 126 GM offspring from a commercial breeding program. We show that both T-DNA copies are present in all our tested GM petunia samples, which underpins the assumption of a single transgenic origin of the undeclared GM petunia. Most likely, the two T-DNAs are integrated in close proximity into the petunia genome. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11240-020-01871-w) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7359168 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-73591682020-07-16 Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia Haselmair-Gosch, Christian Nitarska, Daria Walliser, Benjamin Flachowsky, Henryk Marinovic, Silvija Halbwirth, Heidi Plant Cell Tissue Organ Cult Original Article In 2017, various orange coloured petunia on the market turned out to be genetically modified (GM) without an official authorization for commercialization. Sequence analysis suggested these undeclared plants most probably originated from a plant transformation experiment performed in the 1980s. For a deeper understanding how GM petunia entered classical breeding programmes worldwide, and whether they originated from a single source or not, we undertook a molecular genetic characterization of the T-DNA integration sites in different GM petunia cultivars and breeding lines. By means of genome walking, we isolated different T-DNA sequences, which are located at the junctions between the T-DNA(s) and the petunia DNA. Based on the results obtained we conclude that there are at least two T-DNA copies of different lengths. This is supported by Southern blot analysis. For T-DNA1, the 3′-junction sequence was isolated, whereas the 5′-junction remained unclear. In contrast, for T-DNA2, the 5′-junction sequence was isolated, whereas the sequence isolated from the 3′-region consists only of T-DNA, but did not include the junction from the T-DNA to the petunia DNA. We developed primers for event-specific PCRs and screened a set of three orange GM petunia cultivars and 126 GM offspring from a commercial breeding program. We show that both T-DNA copies are present in all our tested GM petunia samples, which underpins the assumption of a single transgenic origin of the undeclared GM petunia. Most likely, the two T-DNAs are integrated in close proximity into the petunia genome. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11240-020-01871-w) contains supplementary material, which is available to authorized users. Springer Netherlands 2020-06-19 2020 /pmc/articles/PMC7359168/ /pubmed/32684656 http://dx.doi.org/10.1007/s11240-020-01871-w Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Original Article Haselmair-Gosch, Christian Nitarska, Daria Walliser, Benjamin Flachowsky, Henryk Marinovic, Silvija Halbwirth, Heidi Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title | Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title_full | Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title_fullStr | Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title_full_unstemmed | Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title_short | Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petunia |
title_sort | event-specific qualitative polymerase chain reaction analysis for two t-dna copies in genetically modified orange petunia |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359168/ https://www.ncbi.nlm.nih.gov/pubmed/32684656 http://dx.doi.org/10.1007/s11240-020-01871-w |
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