Fenugreek (Trigonella foenum-graecum L.) plays a crucial role in both the food and pharmaceutical industries due to its rich nutritional profile and medicinal properties, such as regulating blood sugar and reducing inflammation. This study offers a comprehensive analysis of the fatty acid composition in fenugreek seeds, revealing significant variability influenced by genetic and environmental factors. The analysis showed that linoleic acid ranged from 32.16 % to 45.96 %, linolenic acid from 17.3 % to 31.21 %, oleic acid from 11.40 % to 20.32 %, stearic acid from 3.78 % to 6.76 %, and palmitic acid from 8.74 % to 16.44 %. Both genotype and environmental conditions had a profound impact on these profiles, with notable variations arising from differences in water availability. The Israel and Kermanshah (Iran) genotypes recorded the highest linoleic acid levels under irrigated conditions, while the Kayseri (Turkey) genotype exhibited the highest linolenic and stearic acid values across both irrigated and non-irrigated environments. Additionally, the Ahvaz (Iran) and Ukraine genotypes excelled in oleic and palmitic acid concentrations. Principal component analysis (PCA) and Additive Main Effects and Multiplicative Interaction (AMMI) biplots further highlighted the complex genotype-environment interactions, providing essential insights for breeding fenugreek varieties optimized for diverse agro-climatic conditions.