Event-specific qualitative polymerase chain reaction analysis for two T-DNA copies in genetically modified orange Petuni
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ORIGINAL ARTICLE
Event‑specific qualitative polymerase chain reaction analysis for two T‑DNA copies in genetically modified orange Petunia Christian Haselmair‑Gosch1 · Daria Nitarska1 · Benjamin Walliser1 · Henryk Flachowsky2 · Silvija Marinovic1 · Heidi Halbwirth1 Received: 5 April 2020 / Accepted: 14 June 2020 / Published online: 19 June 2020 © The Author(s) 2020
Abstract 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. Key message The recently escaped genetically modified orange petunia contain at least two T-DNAs, which have the same integration sites in all tested plant samples, indicating a single transgenic origin. Both T-DNAs can be unambiguously identified via event-specific PCRs. Keywords Petunia × hybrida · Event-specific transgene detection · Anthocyanin · Orange flower colour · Transgenic plant Communicated by Maria Margarida Oliveira. Christian Haselmair-Gosch and Daria Nitarska contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01871-w) contains supplementary material, which is available to authorized users. * Heidi Halbwirth [email protected] 1
Institute of Chemical, Environmental and Bioscience Engineering, Technische Universität Wien, Getreidemarkt 9, 1060 Vienna, Austria
Institute for Breeding Research on Fruit Crops, Julius Kühn-Institut, Pillnitzer Platz 3a, 01326 Dresden, Germany
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Introduction Petunias are economically important balcony and bedding plants, which
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