The present invention relates to Δ8 desaturase genes, which have the ability to convert eicosadienoic acid (EDA; 20:2 ω-6) to dihomo-γ-linolenic acid (DGLA; 20:3 ω-6) and/or eicosatrienoic acid (ETrA; 20:3 ω-3) to eicosatetraenoic acid (ETA; (20:3 ω-3). Isolated nucleic acid fragments and recombinant DNA constructs comprising such fragments encoding Δ8 desaturases along with a method of making long-chain polyunsaturated fatty acids (PUFAs) using these Δ8 desaturases in oleaginous yeast are disclosed.