Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Genetic Transformation of Potato Using the Non-selection Approach

Dmitry Miroshnichenko
* ORCID logo ,
Anna Okuneva
,
Anna Klementyeva
,
Tatiana Sidorova
,
Vadim Timerbaev
ORCID logo ,
Alexander S. Pushin
ORCID logo ,
Sergey V. Dolgov
Version 1 : Received: 8 December 2023 / Approved: 9 December 2023 / Online: 11 December 2023 (06:13:15 CET)

A peer-reviewed article of this Preprint also exists.

Miroshnichenko, D., Klementyeva, A., Sidorova, T., Pushin, A. S., & Dolgov, S. (2024). Genetic Transformation of Potato without Antibiotic-Assisted Selection. Horticulturae, 10(3), 222. Miroshnichenko, D., Klementyeva, A., Sidorova, T., Pushin, A. S., & Dolgov, S. (2024). Genetic Transformation of Potato without Antibiotic-Assisted Selection. Horticulturae, 10(3), 222.

Abstract

The genetic engineering of plants often relies on the use of antibiotic or herbicide resistance genes for initial selection of primary transgenic events. Nevertheless, the commercial release of genetically modified crops containing any marker gene encounters several challenges stemming from the lack of consumer acceptance. Development of strategies enabled the generation of marker-free transgenic plants is a possible alternative to deal with public concerns over the safety of biotech crops. Leveraging the high in vitro regeneration and transformation capabilities of potato, this study examined the factors associated with the development of transgenic plants without presence of selective substances in medium. To compare the antibiotic-assisted with the antibiotic-free transformation, nodal explants from five highly regenerative potato cultivars were inoculated with the Agrobacterium strain AGL0 carrying the pBIN-mGFP5-ER (35SGFP/nosNPTII) plasmid. The efficiency of transient expression and the formation of chimeric and completely transgenic shoots were assessed through vital GFP observations. Under non selective conditions, the regeneration efficiency of stable non-chimeric transgenic shoots varied from 0.9 (‘Chicago’) to 2.7% (#12-36-42). In order to streamline transformation protocol, a specific timeline for collecting transgenic plantlets was established for each analyzed cultivar. This timeline strategy was implemented to generate transgenic potato plants, which demonstrated the suppression of homologous SteIF4Es family genes as a result of the successful introduction of the marker-free hpRNAi construct.

Keywords

Solanum tuberosum L.; antibiotic-free; selectable marker gene; gene silencing; translation initiation factor eIF4E

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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