The oil content and fatty acid composition of berries from two subspecies of sea buckthorn (Hippophaë rhamnoides L.) were investigated. The berries of subsp. rhamnoides contained a higher proportion of oil in seeds (11.3% vs 7.3%, p < 0.01), berries (3.5% vs 2.1%, p < 0.001), and seedless parts (2.8% vs 1.7%, p < 0.01) than the berries of subsp. sinensis. Linoleic (18:2n-6) and alpha-linolenic acids (18:3n-3) comprised about 70% of seed oil fatty acids. Palmitoleic acid (16:1n-7), practically absent in the seed oil, comprised 12.1--39.0% of oil in pulp/peel and 8.9--31.0% of that in the whole berries. More linoleic acid (40.9% vs 39.1%) and less alpha-linolenic acid (26.6% vs 30.6%) was found in the seed oil of subsp. sinensis than in the seed oil of subsp. rhamnoides (p < 0.05). The proportion of palmitoleic acid was higher in the oil of berries of subsp. rhamnoides than the berries of subsp. sinensis (26.0% vs 21.5%, 0.05 < p < 0.1), but was vice versa with alpha-linolenic acid (8.8% vs 11.2%, 0.05 < p < 0.1). The proportions of alpha-linolenic acid correlated inversely with oleic and linoleic acids in the seed oil. In the oil of whole berries, the proportion of palmitoleic acid correlated negatively with the proportions of linoleic and alpha-linolenic acids.