Botrytis cinerea is a model phytopathogenic fungus that infects more than 200 different relevant crops, thus being classified among the top ten fungal plant diseases. B. cinerea exhibits a wide arsenal of tools to infect plant tissues. Most of these factors are related to signal transduction cascades, in which membrane proteins play a key role as a bridge between environmental conditions and intracellular molecular processes. This work describes the first proteomic approach to study the membranome of B. cinerea under different pathogenicity condition induced by using different plant-based elicitors: Glucose and Tomato Cell Wall (TCW). A discovery proteomics analysis of membrane protein extracts was carried out by mass spectrometry. A total of 2,794 proteins were successfully identified, and approximately 46% of them could be classified as membrane proteins based on the presence of transmembrane regions and covalent post-translational modifications. Further analyses showed significant differences in the fungal membranome composition depending on the available carbon source: 804 proteins were exclusively identified when the fungus was cultured in the presence of glucose as sole carbon source, and 251 proteins were exclusively identified in the presence of TCW. Besides, among the 1737 common proteins, a subset of 898 proteins presented clear differences in their abundance and in order to clarify the specific changes produced in the membranome, proteins were categorized according to their gene ontology annotation shown different profiles. As a result of this work, we present here the first description of the membranome of B. cinerea, which will certainly provide a better understanding of this important subproteome in B. cinerea.