| This paper examines the ecotype and organ specific fatty acid content of Echium amoenum, a medicinal plant belonging to the Boraginaceae family in Iran with nutraceutical potential due to polyunsaturated fatty acids (PUFAs) gamma-linolenic acid (GLA, C18:3n-6) and alpha-linolenic acid (ALA, C18:3n-3). 16 ecotypes samples were sampled in different geographical locations (mountainous, coastal, plateau) at similar developmental levels, and lipids in seeds, leaves, and roots were extracted in n-hexane using Soxhlet, separated by thin-layer chromatography (TLC; Rf GLA = 0.26; methyl ester= 0.54), and quantified using gas chromatography (GC-FID; Agilent 7890B, DB-23 column), and analyzed through hierarchical cluster analysis. The highest oil content (18.3-24.7%, E5 ecotype max. 24.7) and dominance of PUFA: ALA 39.4-48.3% (K9: 46.25%), linoleic acid 22.9, GLA 3.1-4.9% (So16: 4.9%), and PUFA: SFA ratio of over 9: 1. The oil, ALA, and GLA content of leaves (moderate 0.89%), roots (low 0.6%), SFA/MUFA presence (palmitic 6.38%), and GLA (0.08-0.89) and no Oleic, respectively. HCA showed that there were four clusters, the first group with High-GLA includes: Galougah (P2), the second group consists of the ecotypes of Isfahan (S6), the third group comprises the ecotypes of Sochalma (So16) while the fourth group categorizes the three ecotypes of Behshahr (B8), Dohezar (2000), and Lorestan (L15). With bootstrap (>85% reproducibility) showing environmental modulation of desaturase pathways (D6/D15). TLC-AutoCAD densitometry had a strong correlation with GC (r=0.92, p<0.01), which confirmed organ-specific partitioning in favor of seeds to extract GLA/ALA. Selective breeding and cultivation leveraging ecotypic variation: So16 is better for GLA, anti-inflammatory supplements, borage, and evening primrose oil substitute. The TLC-GC-HCA model provides a strong, cost-efficient system of PUFA profiling in medicinal plants to combine lipid biochemistry, chemotaxonomy, and sustainable nutraceutical use. |