Skip to main content
Log in

Generation of stable transgenic Brassica napus cv. Jet Neuf cell cultures as a tool to investigate in planta protein function

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript


Analysis of proteins often requires generation of transgenic plants to gain knowledge of their in planta function. The acquisition of such plants represents a significant investment in terms of workload, time, and space. Here we describe an easy-to-use alternative system by transforming Brassica napus cv. Jet Neuf suspension cells derived from microspores. The cells can be transformed with high efficiency, and even double transformations with different transgenes are possible. In addition, we describe a procedure that allows for long-term storage to maintain diverse transgenic cell lineages with low effort, to build up cell suspension libraries for research purposes. Overall, the work can be helpful for scientists that want to use B. napus as a research organism in a reduced time-frame without the need for generating whole plants and the requirement of larger greenhouse spaces.

Key message

This study establishes a protocol for generation of stable Brassica napus transgenic cell cultures, their long-term storage, and contains examples how these cell cultures can be used in studying gene function in planta without generating whole plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The datasets of the current study are presented in this article or are available upon a reasonable request.


Download references


We would like to thank Prof. Thomas Clemente, Center for Plant Science Innovation, Department of Agronomy and Horticulture, University of Nebraska-Lincoln, for providing the RD29a promoter. We also would like to thank Dr. John Shanklin, Brookhaven National Laboratory, NY, for providing the stock of B. napus cv. Jet Neuf cell cultures used in this work, and WSU’s Franceschi Microscopy and Imaging Center for providing access to a confocal microscope and supporting undergraduate research.


This project was supported by an Agriculture and Food Research Initiative competitive grant 2019-67013-29160 of the USDA National Institute of Food and Agriculture (NIFA) to H.H. WSU financially supported J.L. in his undergraduate research studies through a CAS Summer Fellowship.

Author information

Authors and Affiliations



J.L. is an undergraduate student of WSU’s Honors College who established the transformation and long storage procedures, and also performed the GUS stainings shown in this work. S.M. did the RT-qPCR analysis, S.S. did salt and low-sucrose with salt assays, M.K. did the confocal analysis, and R.A-S. generated the proESR1:GUS and proRD29a:BrRAP2.4-1 expression constructs. H.H. is the principal investigator, designed the figures, and developed the manuscript together with J.L. All authors contributed with editing and agreed on the manuscript.

Corresponding author

Correspondence to Hanjo Hellmann.

Ethics declarations

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Additional information

Communicated by Heidi Halbwirth.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary Material 1

Supplementary Material 2

Supplementary Material 3

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Landers, J., Mooney, S., Smalley, S. et al. Generation of stable transgenic Brassica napus cv. Jet Neuf cell cultures as a tool to investigate in planta protein function. Plant Cell Tiss Organ Cult 154, 633–643 (2023).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: