1. The unit offers Agrobacterium-mediated tissue-culture transformation of potato, tobacco and tomato ssp.
2. PEG-mediated transformation of protoplasts from cell cultures or root cells (Arabidopsis).
3. Maintenance of our Col-0 cell culture (wild type).

Examination of the protoplat transformation efficiency by flow cytometric anaylsis (FCA).

Protoplasts isolated from root cell culture are transformed with a 35S::GFP construct (12kb) as a routine control. As an alternative to calculating the transformation efficiency, we have used FCA as a means for detecting cells expressing GFP. The negative population is marked in black. Cells that are strongly expressing GFP have a fluoresence signal that is well above that of the total plant cell population‘s green autofluoresence (cells marked in dark green); cells marked in light green are those with weaker GFP fluoresence emission. In this example 23% of the cells were transformed with GFP, and 50% of those are strongly expressing GFP.

Current Publications

2022

(publication) The tomato resistance gene Bs4 suppresses leaf watersoaking phenotypes induced by AvrHah1, a transcription activator-like effector from tomato-pathogenic xanthomonads. Schenstnyi K, Strauß A, Dressel A, Morbitzer R, Wunderlich M, Andrade AG, Phan TT, Aguilera PLA, Brancato C, Berendzen KW, Lahaye T. New Phytol. 2022 Sep 2. doi: 10.1111/nph.18456.

2019

(publication) ARR22 overexpression can suppress plant Two-Component Regulatory Systems. Wallmeroth N, Jeschke D, Slane D, Nägele J, Veerabagu M, Mira-Rodado V, Berendzen KW. PLoS One. 2019 Feb 11;14(2):e0212056. doi: 10.1371/journal.pone.0212056. eCollection 2019.

2015

(publication) An RLP23-SOBIR1-BAK1 complex mediates NLP-triggered immunity. Albert I, Böhm H, Albert M, Feiler CE, Imkampe J, Wallmeroth N, Brancato C, Raaymakers TM, Oome S, Zhang H, Krol E, Grefen C, Gust A, Chai J, Hedrich R, Van den Ackerveken G, Nürnberger T (2015). Nature Plants, 15140, doi: 10.1038/NPLANTS.2015.140.

(publication)  Development of a tomato plant resistant to Clavibacter michiganensis using the endolysin gene of bacteriophage CMP1 as a transgene. J. Wittmann, C. Brancato, K. W. Berendzen and B. Dreiseikelmann (2015). Plant Pathology DOI: 10.1111/ppa.12417

Transformation Unit Protocols

Cell Culture 01.doc
Cell Culture Maintenance (Arabidopsis Col-0)

BY-2-protopl.doc
BY-2 Tobacco Cell Suspension Protoplasts
(NOTE: we no longer maintain BY-2 suspension cells or culture since 2020).

PEG transfection protocol 2018

Arabodopsis Cell Culture Protoplasts
 

REFERENCING

Please cite for the standard protoplast transfection protocol after Feb. 2017:

Mehlhorn D.G., Wallmeroth N., Berendzen K.W., Grefen C. (2018) 2in1 Vectors Improve In Planta BiFC and FRET Analyses. In: Hawes C., Kriechbaumer V. (eds) The Plant Endoplasmic Reticulum. Methods in Molecular Biology, vol 1691. Humana Press, New York, NY.

Please cite for the 96-well transfection protocol:

Berendzen KW, Böhmer M, Wallmeroth N, Peter S, Vesić M, Zhou Y, Tiesler FK, Schleifenbaum F, Harter K. Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry. Plant Methods. 2012 Jul 12;8(1):25.

Transfections performed before Feb.2017 cite:

Schütze K, Harter K, Chaban C (2009) Bimolecular fluorescence complementation (BiFC) to study protein-protein interactions in living plant cells. Methods Mol Biol 479: 189-202.

STABLE TRANSFORMATIONS

PROTOCOLS

PotatoStabile.pdf
Stable Transformation protocol for potato

Transformation protocol for tobacco

Protocol for tomato stable transformation
Protocol for tomato stable transformation

LotusAgroTrafo1.pdf
Protocol for Lotus stable transformation

REFERENCING

Please cite for tomato transformations:
Wittmann et al. 2015. Plant Pathology. Doi: 10.1111/ppa.12417
http://onlinelibrary.wiley.com/doi/10.1111/ppa.12417/abstract

OTHER PROTOCOLS

Protoplasts isolation from leaves of Arabidopsis thaliana