Tomato (Solanum lycopersicum) is an important fruit and vegetable crop in worldwide. The fertility of tomato reproductive organs can be dramatically decreased when ambient temperatures rise above 35 °C, reducing tomato fruit yield. It is necessary to identify transcription factors (TFs) and target genes involved in heat stress response (HSR) signaling cascades in tomato flower buds to improve tomato plant thermotolerance. In this study, we profiled genes expressed in three developmental stages of tomato flower buds. Red and turquoise modules for heat stress (HS) were identified through gene co-expression network analysis, and the genes within these modules were enriched in HS-related pathways. By focusing on the TFs in the two modules, we identified several novel HSR-related TFs, including SlWRKY75, SlMYB117, and SlNAM. Furthermore, homology analysis illustrated a conserved signaling cascade in tomato. Lastly, we identified and experimentally validated four HSF-regulated genes, namely SlGrpE, SlERDJ3A, SlTIL, and SlPOM1, that likely modulate thermotolerance in plants. These results provide a high-resolution atlas of gene expression during tomato flower bud development under HS conditions, which is a valuable resource for uncovering potential regulatory networks associated with the HSR in tomato.

番茄（Solanum lycopersicum）是世界范围内重要的果蔬作物。当环境温度升至35°C以上时，番茄生殖器官的生育力会急剧下降，从而降低番茄果实产量。有必要鉴定番茄花芽中参与热应激反应（HSR）信号级联的转录因子（TF）和靶基因，以提高番茄植株的耐热性。在这项研究中，我们分析了番茄花芽三个发育阶段表达的基因。通过基因共表达网络分析鉴定了热应激（HS）的红色和青绿色模块，这些模块内的基因在热应激相关通路中富集。通过关注这两个模块中的 TF，我们确定了几个新颖的 HSR 相关 TF，包括 SlWRKY75、SlMYB117 和 SlNAM。此外，同源性分析表明番茄中存在保守的信号级联。最后，我们鉴定并通过实验验证了四个 HSF 调节基因，即 SlGrpE、SlERDJ3A、SlTIL 和 SlPOM1，它们可能调节植物的耐热性。这些结果提供了 HS 条件下番茄花芽发育过程中基因表达的高分辨率图谱，这是揭示番茄 HSR 相关潜在调控网络的宝贵资源。