Development of Real-time PCR Assays for the Detection of the pin II Terminator (tpinII) Used in GM Constructs and Its Donor Organism, Potato (Solanum tuberosum)

Abstract

Two singleplex TaqMan methods were developed for the detection of potato targets: one for the detection of the tpinII terminator, which is an emerging terminator used in GM constructs, and one for the detection of the endogenous StLS gene of potato. Performance criteria such as specificity and sensitivity were successfully tested for the two methods, taking into account the recommendations of international guidelines. The presence of the StLS target was checked in 16 potato cultivars. The StLS target is present at low copy number and can be used for quantitation purposes, as demonstrated on transgenic potatoes in this paper. The StLS target is an excellent candidate to replace the presently recommended endogenous target based on the UGP gene, which shows several disadvantages due to its high copy number and lack of specificity. The research also indicates that DNA can easily be extracted from different parts of potato tubers with a classical cetyltrimethylammonium bromide method.

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Acknowledgements

We are very grateful to Frédéric Gastiny of the University of Liège (Gembloux Agro-Bio Tech, Belgium) for providing the GM potato lines and Yordan Muhovski of CRA-W for his kind help with Southern blots.

Funding

The StLS work was carried out thanks to a grant from the Belgian Federal Science Policy Office. The development of the tpinII was funded by the Belgian Federal Public Service for Health, Food Chain Safety and Environment through the contract UGMMONITOR (convention RF 11/6242).

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Correspondence to Frederic Debode.

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Frederic Debode declares that he has no conflict of interest. Kamila Zdeňková declares that she has no conflict of interest. Eric Janssen declares that he has no conflict of interest. Anette Tizolova declares that she has no conflict of interest. Patrick du Jardin declares that he has no conflict of interest. Gilbert Berben declares that he has no conflict of interest. Kateřina Demnerova declares that she has no conflict of interest.

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This article does not contain studies with human or animal subjects. Samples used are food samples.

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Debode, F., Zdeňková, K., Janssen, E. et al. Development of Real-time PCR Assays for the Detection of the pin II Terminator (tpinII) Used in GM Constructs and Its Donor Organism, Potato (Solanum tuberosum). Food Anal. Methods 11, 2172–2180 (2018). https://doi.org/10.1007/s12161-018-1203-4

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Keywords

  • tpinII
  • Screening
  • Potato detection
  • Endogenous target
  • GMO