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Heine, P.A.* ; Nosenko, T. ; Kistner, S.* ; Oliphant, K.D.* ; Hanke-Uhe, M.* ; Shahid, A.* ; Hu, B.* ; Kucklick, M.* ; Lehmler, N.* ; Becker, M.* ; Goerke, N.* ; Korn, J.M.* ; Linke, T.* ; Meier, D.H.* ; Perl, A.* ; Polten, S.* ; Priess, V.* ; Schäckermann, D.* ; Schubert, M.* ; Schumacher, J.* ; Winkler, J.B. ; Engelmann, S.* ; Rennenberg, H.* ; Schnitzler, J.-P. ; Dübel, S.* ; Hust, M.* ; Hänsch, R.* ; Kaufholdt, D.*

Phage display derived antibodies against antimicrobial peptide FsPDF2 reveal stress response in European beech.

Plant Biotechnol. J., DOI: 10.1111/pbi.70431 (2025)
Verlagsversion Forschungsdaten DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Plant defensins (PDFs) are cysteine-rich antimicrobial peptides (AMPs) that are important components of plant immunity. They occur constitutively in various plant tissues but are also upregulated upon stress. Therefore, these molecules are of great interest as markers for the diagnosis of early forest stress response in plants at the molecular level. PDFs are small peptides (~5 kDa) with a compact tertiary structure, requiring specific protocols and dedicated antibodies for detection by quantitative ELISA. We developed monoclonal recombinant antibodies using phage display in solution against the correctly folded antigen defensin FsPDF2 from beech (Fagus sylvatica) and analysed the antibody-antigen interaction in silico with AlphaFold 3. In a proof-of-principle study, we investigated the FsPDF2 stress response to abiotic (drought) and biotic (gall midge) stresses. Notably, we established an assay for defensin quantification in crude plant extract, detecting for the first time natively folded proteins in a specific sandwich ELISA. Our antibody generation strategy can be transferred by practitioners to other small antimicrobial peptides (AMP), paving the way to study this group of proteins and their corresponding stress response comprehensively.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Amp Quantification ; Antimicrobial Peptides ; Crude Plant Extract Elisa ; Drought/biotic Stress ; Forest Stress Diagnostic ; Phage Display Recombinant Antibodies ; Plant Defensins
ISSN (print) / ISBN 1467-7644
e-ISSN 1467-7652
Zeitschrift Plant Biotechnology Journal
Verlag Wiley
Begutachtungsstatus Peer reviewed
Institut(e) Research Unit Environmental Simulation (EUS)