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1-deoxy-D-xylulose-5-phosphate synthase (DXS) is a rate-limiting enzyme in terpene synthesis that can affect the accumulation of secondary metabolites in plants. In this study, three <i>DXS</i> gene family members were identified in the tomato genome-wide database. Using bioinformatics methods, we analyzed the gene structure, evolutionary affinities, and cis-acting elements of the <i>SlDXS</i> gene family members. Promoters of <i>SlDXS</i> genes contain plant hormone-responsive elements such as the CGTCA-motif, TGACG-motif, ABRE, TCA-element, TGA-element, ERE, CAT-box, and AACA-motif, which suggested that the <i>SlDXS</i> gene family may play an important role in hormone response. The RT-qPCR analysis showed that the tomato <i>DXS2</i> gene was able to respond upon exposure to methyl jasmonate (MeJA). The construction of a virus-induced gene silencing (VIGS) vector for the <i>SlDXS</i> gene showed that the <i>SlDXS2</i> gene was also able to respond to MeJA in silenced plants, but the induction level was lower relative to that of wild-type plants. The <i>SlDXS1</i> gene is associated with the synthesis of photosynthetic pigments. This study provides a reference for the further elucidation of the <i>DXS</i> gene’s biological function in the terpenoid synthesis pathway in tomatoes.