Every year, viticulture faces several disease outbreaks caused by pathogens with different life cycles and modes of action, and due to climate changes, the appearance of new pathogens is a concern for viticulture. To cope with the different pathogenic microorganisms, the plant must recognise the invading agent and arm itself with an arsenal of defence reactions, including the accumulation of antimicrobial metabolites.

The modulation of plant metabolites is one of the first responses to biotic factors. Their rapid synthesis can greatly contribute to strengthening plant defence allowing it to adapt, minimise pathogen colonisation and survive. Despite the scientific community's efforts to characterise the grapevine defence responses to pathogens, the molecules involved in pathogen control and their specific localisation remain to be deciphered.

Mass Spectrometry Imaging (MSI) analytical techniques enable one to visualise and map the spatial distribution of numerous biological molecules within plant tissues allowing a better understanding of their biosynthesis, localisation and function.

In this review, we explore the applications of MSI techniques to the study of plant/microbe interactions focusing on grapevine studies. This review opens new doors to gain a comprehensive understanding of the dynamics and variations of metabolite profiles in grapevine organs, at different developmental stages and under various stress conditions. This knowledge is crucial for elucidating the role of specific metabolites in grapevine defence mechanisms, and for identifying specific regions of high or low metabolite production, which can contribute to targeted breeding to improve disease resistance traits and have an impact on grapevine productivity and quality.