Extracellular ATP and Metabolomic Fingerprints of Regulated Cell Death in Tomato and Arabidopsis
Authors
Jo Petter Helstad (presenting author) [1, 2]
Daniela Jorgelina Sueldo [1]
Gaston Courtade [2]
Leah Wanja Gachao [1]
Susanne Hansen Troøyen [2]
Keywords
Cell death; NMR metabolomics; Extracellular ATP; Tomato
Abstract
Regulated cell death (RCD) is critical for plant development, differentiation, and responses to environmental stress. Environmentally induced RCD can be triggered by abiotic and biotic factors to remove damaged cells or limit pathogen infection. Heat stress is an example of an abiotic factor in which increased temperatures trigger RCD. Dying cells release damage associated molecular patterns (DAMPs), including nucleotides like adenosine triphosphate (ATP). Extracellular ATP (eATP) can be recognized by neighboring cells to initiate stress responses or amplify RCD cascades. Ion leakage measurements can estimate RCD, as dying cells lose membrane integrity, leading to the release of cellular electrolytes. Our lab has established a protocol to estimate quantitative RCD after heat shock in Arabidopsis thaliana seedlings (Gacho & Sueldo, 2025). Here, we adapt this protocol to assess heat shock-induced RCD in tomato (Solanum lycopersicum) leaves using electrolyte leakage, showing that the protocol is easy to adapt to other species. To continue, we infiltrated A. thaliana seedlings and tomato leaves with external ATP and observed increased electrolyte leakage. This suggests eATP as a potential signal molecule involved in RCD in plants. Finally, molecules released during RCD can act as a metabolomic fingerprint of stress responses. We are employing nuclear magnetic resonance (NMR) spectroscopy to characterize the metabolomic fingerprint of RCD in A. thaliana seedlings and tomato leaves upon heat stress and external ATP infiltration. Our preliminary data suggests that it is possible to characterize heat stressed induced RCD with NMR metabolomic fingerprinting.
Funding
No funding information was provided in the submission.