Metabolomic profiling of grapevine leaf apoplast upon pathogen infection

Abstract

Grapevine is one of the most important crops worldwide. In a climate change scenario, vineyards face not only abiotic challenges, but also recurrent annual outbreaks of diseases, such as downy mildew and grey mould, caused by the oomycete Plasmopara viticola (PV) and the fungi Botrytis cinerea (BC), respectively. By regulating several metabolic pathways, the grapevine can defend itself and survive from pathogens attack.

In our previous studies [1,2], upon PV and BC infection, our results showed that grapevine’s metabolic dynamics has a pivotal role on the activation/shutdown of specific defence pathways. Flavonoids, carbohydrates, and lipids are the metabolic classes with higher modulation, exhibiting distinct temporal regulation patterns. Further analysis, using mass spectrometry imaging, revealed that stilbene phytoalexins and carbohydrates strongly accumulate near PV and BC infection sites, with levels determined by both post-infection timing and molecular complexity [3,4].

These observations raise the possibility that specific metabolites within these classes are not only responsible for Vitis species’ tolerance/susceptibility, but also play an active role in pathogen recognition and plant defence mechanisms.

In this context, exploring the metabolic content of grapevine leaves apoplast (APF) is essential, as this compartment is the first hub of plant-pathogen communication playing a key role in pathogen recognition, activation of signalling cascades, and pathogen counterattack. Preliminary team studies [5] at the constitutive level revealed a broad spectrum of metabolic classes, being the most represented ones ‘Lipids’, followed by ‘Phenolic compounds’ and ‘Carbohydrates; however, the metabolic characterisation of the APF during grapevine-pathogen interactions has never been reported.

To gain a better understanding of these metabolic dynamics, we performed a metabolic characterisation of grapevine leaves APF following infection with PV and BC, using ultra-high-resolution mass spectrometry. In this study, we integrated intracellular and extracellular (apoplast) untargeted metabolomics data to provide a comprehensive view of grapevine’s metabolic response to pathogen invasion, with the goal of identifying crucial metabolites involved in grapevine’s defence. This knowledge is fundamental for elucidating the role of metabolites in grapevine defence and paves the way for new sustainable disease control strategies.

References

[1] Nascimento et al. 2019. Plant. Physiol. Biochem., 137, 1–13. DOI:10.1016/j.plaphy.2019.01.026.

[2] Cavaco et al. 2021. Plant. Physiol. Biochem., 163, 230–238. DOI:10.1016/j.plaphy.2021.04.001

[3] Maia et al. 2022. Front. Plant Sci., 13:1012636. DOI:10.3389/fpls.2022.1012636

[4] Maia et al. 2023. J. Agric. Food Chem., 71, 42, 15569–15581. DOI:10.1021/acs.jafc.3c03620

[5] Figueiredo et al. 2021. Physiol. Plant., 171: 343–357. DOI:10.1111/ppl.13198.

Acknowledgements

The authors acknowledge the financial support from Fundação para a Ciência e a Tecnologia (FCT-Portugal) through Marisa Maia research contract 2022.07433.CEECIND (DOI: 10.54499/2022.07433.CEECIND/CP1715/CT0009), the DEVINE-ME exploratory project 2023.14736.PEX (DOI: 10.54499/2023.14736.PEX) and the Centre grant to BioISI UID/04046/2025 (DOI: 10.54499/UID/04046/2025), to LEAF under the projects UID/AGR/04129/2025 (DOI: 10.54499/UIDB/04129/2020) and UIDP/04129/2025 (DOI: 10.54499/UIDP/04129/2020) and Associate Laboratory TERRA LA/P/0092/2020 (DOI:10.54499/LA/P/0092/2020). Ana Cruz-Silva fellowship was funded by the BioSys2 PhD program (UI/BD/153050/2022). Support from the MassLor research infrastructure at the University of Lorraine is also acknowledged.