Received: November 11th, 2021 ; Accepted: January 30th, 2022 ; Published: March 17th, 2022 ; Correspondence: linda.legzdina@arei.lv Lipids are a minor nutritional component of barley (Hordeum vulgare L.) grain and have not been as widely explored as the major components. The aim of this study was to investigate the effect of genotype and environment, including conventional farming system with three crop management regimes, differing in agrochemical input, and organic farming system, on oil content and fatty acid composition in grain of two covered and four hulless spring barley genotypes during two growing seasons. Genotype significantly affected oil content and it was on average 4.26% and ranged in individual barley samples from 2.87 to 5.53%. We found linoleic, oleic, palmitic, α-linolenic, stearic and capric fatty acids in average proportions of 55.6; 21.3; 18.6; 3.7; 0.6 and 0.4%, respectively. Higher average oil content and proportion of α-linolenic acid was found in covered barley. Crop management regime did not significantly affect oil content but had some effect on the proportion of linoleic, α-linolenic, oleic and stearic acid. Decrease of chemical inputs was in favour of oil content and proportion of α-linolenic, oleic and stearic acids but did not promote linoleic acid. Waxy hulless barley line with high oil content and a very high proportion of linoleic acid was identified.