Evolutionary and functional insights into HSF gene family underpin thermal adaptation in the plateau plant Anisodus tanguticus

Anisodus tanguticus, a plateau-dwelling medicinal plant endemic to high-altitude regions, synthesizes pharmaceutically critical tropane alkaloids but faces escalating threats from climate warming. While its unique thermal adaptation mechanisms remain enigmatic, heat shock transcription factors (HSFs) are hypothesized to orchestrate stress resilience in this species. Here, we present a genome-wide evolutionary and functional dissection of the HSF gene family in A. tanguticus, identifying 20 HSF members (AntHSFs) with distinct structural and regulatory features. Phylogenetic reconstruction classified AntHSFs into three canonical subfamilies, revealing lineage-specific diversification patterns shaped by tandem and segmental duplication events. Conserved motif architectures and DNA-binding domains underscored functional divergence, while synteny analysis highlighted evolutionary constraints and adaptive innovations compared to model species. The transcript dynamics analysis under heat treatment revealed the dynamic changes in enzyme activity and gene expression of the MDA, GSTT, GSTF, and Cu/ZnSOD genes under different heat-treated times. The AntHSF gene exhibited stage-specific expression patterns consistent with antioxidant enzyme activity, indicating that the AntHSF family plays a critical role in coordinating the antioxidant gene response through temporal regulation networks. Notably, evolutionary analysis and sub-cellular localization revealed that AntHSF7 and AntHSF9 were potentially linked to plateau-specific adaptation. This study deciphers the evolutionary drivers and functional specialization of HSF genes in A. tanguticus, providing mechanistic insights into its thermal resilience and a genetic roadmap for conserving and improving this climate-vulnerable medicinal species.