AbstractLettuce (Lactuca sativa L.) is an important leafy vegetable consumed worldwide. Heat‐induced bolting and flowering greatly limit lettuce production during the summer. Additionally, MADS‐box transcription factors are important for various aspects of plant development and architecture (e.g., flowering and floral patterning). However, there has been no comprehensive study of lettuce MADS‐box family genes. In this study, we identified 82 MADS‐box family genes in lettuce, including 23 type I genes and 59 type II genes. Transcriptome profiling revealed that LsMADS gene expression patterns differ among the various floral stages and organs. Moreover, heat‐responsive cis‐elements were detected in the promoter regions of many LsMADS genes. An in situ hybridization assay of 10 homologs of flower‐patterning genes and a yeast two‐hybrid assay of the encoded proteins revealed that the ABC model is conserved in lettuce. Specifically, the APETALA1 (AP1) homolog in lettuce, LsMADS55, is responsive to heat and is specifically expressed in the inflorescence meristem and pappus bristles. The overexpression of LsMADS55 results in early flowering in Arabidopsis thaliana. Furthermore, we observed that the heat shock factor LsHSFB2A‐1 can bind to the LsMADS55 promoter in lettuce. Therefore, a model was proposed for the LsMADS‐regulated floral organ specification and heat‐induced flowering in lettuce.