Post-translational modification of proteins through lysine succinylation plays important regulatory roles in living cells. Lysine succinylation was recently identified as a novel post-translational modification in several organisms, including Escherichia coli, yeast, Toxoplasma gondii, and HeLa cells, as well as mouse liver. Interestingly, few sites of lysine succinylation were detected in plants. In this study, we identified 347 sites of lysine succinylation in 202 proteins of tomato by using high-resolution mass spectrometry.