ORIGINAL RESEARCH article

Front. Plant Sci.

Sec. Technical Advances in Plant Science

Volume 16 - 2025 | doi: 10.3389/fpls.2025.1623387

Genetic transformation of Primula sieboldii using Agrobacterium rhizogenes and whole-plant regeneration from transgenic hairy roots

Provisionally accepted
  • 1University of Potsdam, Potsdam, Germany
  • 2Institute for Breeding Research on Horticultural Crops, Julius Kühn Institute - Federal Research Centre for Cultivated Plants, Quedlinburg, Germany

The final, formatted version of the article will be published soon.

Primula sieboldii E. Morren is a widely cultivated ornamental plant with horticultural and pharmacological value. However, the lack of a developed transformation system has limited genetic studies and biotechnological applications of this species. In this study, we established a transformation method using Agrobacterium rhizogenes for the genetic manipulation of Primula sieboldii. The protocol consists of two stages: initial hairy root transformation and subsequent wholeplant regeneration from transgenic hairy roots through somatic embryogenesis. Comprehensive molecular analyses confirmed the stable integration and expression of various transgenes from the root-inducing plasmid (Ri) and the binary vector carrying the RUBY reporter in independent transgenic lines, as well as the stable germline transmission of the transgene to progeny. The protocol is effective, with 5% of treated explants successfully forming transformed hairy roots expressing the RUBY reporter, from which regenerating into transgenic plants were regenerated. The established method provides a valuable tool for genetic and molecular studies of heterostyly and the selfincompatibility system in the genus Primula, while also offering practical applications in molecular breeding and plant biotechnology. Additionally, hairy root cultures provide a platform for metabolic engineering and the exploration of biologically active secondary metabolites with pharmacological applications.* The number of explants producing white hairy roots is not equal to the total number of explants producing hairy roots, since one explant can produce red and white hairy roots, only red, or only white.

Keywords: Primrose, Genetic Engineering, plant biotechnology, plant tissue culture, Agrobacterium-mediated plant transformation

Received: 05 May 2025; Accepted: 01 Jul 2025.

Copyright: © 2025 Gad, Plaschil, Kappel, Lenhard and Huu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Karol Gad, University of Potsdam, Potsdam, Germany

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