The current study compared beef production, quality and fatty acid (FA) profiles of yearling steers fed a control diet containing 70 : 30 red clover silage (RCS) : barley-based concentrate, a diet containing 11% sunflower seed (SS) substituted for barley, and diets containing SS with15% or 30% wheat dried distillers’ grain with solubles (DDGS). Additions of DDGS were balanced by reductions in RCS and SS to maintain crude fat levels in diets. A total of two pens of eight animals were fed per diet for an average period of 208 days. Relative to the control diet, feeding the SS diet increased (P<0.05) average daily gain, final live weight and proportions of total n-6 FA, non-conjugated 18:2 biohydrogenation products (i.e. atypical dienes) with the first double bond at carbon 8 or 9 from the carboxyl end, conjugated linoleic acid isomers with the first double bond from carbon 7 to 10 from the carboxyl end, t-18:1 isomers, and reduced (P<0.05) the proportions of total n-3 FA, conjugated linolenic acids, branched-chain FA, odd-chain FA and 16:0. Feeding DDGS-15 and DDGS-30 diets v. the SS diet further increased (P<0.05) average daily gains, final live weight, carcass weight, hot dressing percentage, fat thickness, rib-eye muscle area, and improved instrumental and sensory panel meat tenderness. However, in general feeding DGGS-15 or DDGS-30 diets did not change FA proportions relative to feeding the SS diet. Overall, adding SS to a RCS-based diet enhanced muscle proportions of 18:2n-6 biohydrogenation products, and further substitutions of DDGS in the diet improved beef production, and quality while maintaining proportions of potentially functional bioactive FA including vaccenic and rumenic acids.