Six wether sheep with ruminal and duodenal cannulas were used in a replicated 3 × 3 Latin square to determine the effect of rye grain (from an open-pollinated or a hybrid cultivar) and maize grain from hybrid cultivar inclusion in the diet on ruminal fermentation parameters and nutrient digestion in different gastrointestinal tract compartments. The experimental diets consisted of meadow hay (800 g/day) and a concentrate mixture (300 g/day) that contained rye grain from an open-pollinated cultivar (OPRG), rye grain from a hybrid cultivar (HRG), or maize grain from a hybrid cultivar (MG) as a main energy source. Each experimental period lasted 17 days and consisted of 10 days for diet adaptation and 7 days for data and sample collection. Ruminal fermentation (pH, short-chain fatty acids, and ammonia concentration) and nutrient digestion up to the duodenum (in reticulorumen, omasum, and abomasum), in the intestine, as well as in the total digestive tract was investigated. Mean ruminal pH was higher for MG compared to HRG and OPRG (P<0.01) but did not differ between HRG and OPRG (6.45, 6.35, and 6.29 for MG, HRG, and OPRG, respectively). Treatment did not affect short-chain fatty acid concentration in the rumen, except for the molar proportions of valeric acid and of isovaleric acid of the total short-chain fatty acids, which was or tended to be higher (P≤0.06) for MG compared to HRG and OPRG. Less starch was digested in the gastrointestinal tract up to the duodenum and more in the intestine for MG compared to HRG and OPRG (P=0.01); however, total tract nutrient digestibility did not differ between treatments. In conclusion, usage of rye grain in sheep diets resulted in a lower ruminal pH compared to maize grain usage. No differences for ruminal fermentation, nutrient digestion up to the duodenum, in the intestine, as well as in the total digestive tract between rye grain from open-pollinated and hybrid cultivar usage in sheep diets were found.