<p>To elucidate the phytobiochemical mechanisms underlying differentiation among wild populations of <i>Tithonia diversifolia</i> (Helms) A. Gray and establish how soil parameters regulate metabolic pathways. Methods. 90 individuals from three populations (Ixtaczoquitlán, Orizaba, Rafael Delgado; Veracruz, Mexico) were analyzed. A multi-analytical approach included lipid profiling by gas chromatography–mass spectrometry (GC–MS), identification of secondary metabolites via HPTLC, quantitative bromatological analyses, photosynthetic pigment quantification, and comprehensive edaphoclimatic characterization. Statistical modeling integrated soil chemistry, climatic dynamics, and phytobiochemical responses. Results. Populations exhibited distinct metabolic phenotypes shaped by edaphic stress. Plants from Rafael Delgado expressed a classical hormetic response under moderate stress (neutral pH, high EC and CEC, low organic matter, clayey soil), with upregulation of biosynthetic pathways resulting in higher protein content (27.25 ± 1.12% DW) and a diverse fatty acid profile (seven compounds). In contrast, Ixtaczoquitlán and Orizaba populations, under more favorable soils, maintained homeostatic regulation prioritizing primary metabolism, with higher chlorophyll accumulation (1.96 ± 0.10&#xa0;mg&#xa0;g⁻¹) but reduced synthesis of defensive compounds. Foliar pH remained stable (6.7 ± 0.3) across sites, suggesting a robust self-regulation capacity despite edaphoclimatic variability. Conclusions. Stress-induced metabolic switching emerges as a key adaptive mechanism in this non-model species, highlighting how environmental gradients reprogram biosynthetic pathways. Hormesis-driven enhancement of bioactive compounds positions <i>T. diversifolia</i> as a promising system for biotechnology aimed at stress-induced biocompound production. These findings advance the state of the art in plant metabolic plasticity and support the sustainable exploitation of renewable ethnobotanical resources.</p>