The phytohormone abscisic acid (ABA) plays a central role in organizing adaptive responses in plants to various abiotic stresses, helping the plant minimize the negative impact on growth and development. Rapid and direct detection of ABA is valuable for investigating plant responses to abiotic stress. In this work, we propose a novel label-free, non-competitive immunoassay for detecting and quantifying ABA easily and rapidly using a surface plasmon resonance (SPR) biosensor. The SPR sensor chip was functionalized with a commercial anti-ABA antibody, characterized for its affinity, binding kinetics, and specificity using the same platform. The direct assay demonstrated high specificity and sensitivity, with a calculated limit of detection of 1.36 ng/mL in buffer. The new immunosensor was applied to determine ABA concentrations directly in xylem sap samples from tomato plants subjected to abiotic stress (drought and high salinity) and was able to accurately reflect ABA levels corresponding to the applied stress. The results were comparable to the reference method, ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS), establishing this new immunosensor as a novel detection method for rapid and reliable monitoring of ABA levels associated with abiotic stress in tomato plants.

Biosensors Group Lammertyn Lab KU Leuven Willem de Croylaan 42 3001 Leuven Belgium

KU Leuven Plant Institute Kasteelpark Arenberg 31 3001 Leuven Belgium

Laboratory of Growth Regulators Institute of Experimental Botany of the Czech Academy of Sciences Palacký University Šlechtitelů 27 783 71 Olomouc Czech Republic

Molecular Plant Hormone Physiology Lab KU Leuven Willem de Croylaan 42 3001 Leuven Belgium

Molecular Structural and Translational Virology Rega Institute KU Leuven Herestraat 49 3000 Leuven Belgium

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