The objective of this study was to determine, under the soil and climatic conditions of Southern Chile, the effect of the ploidy of perennial ryegrass (Lolium perenne L.) cultivars on herbage production, nutritive value, grazing preference and utilization of pasture produced. This study was conducted in southern Chile, Valdivia Province, and was evaluated for 3 years. The tetraploid cultivars used were Quartet (4n), Gwendal (4n), Pastoral (4n) and Napoleon (4n). The diploid cultivars were Anita (2n), Jumbo (2n), Aries (2n), and Yatsyn 1 (2n). When the average sward height reached 20 cm, all plots were simultaneously grazed by dairy cows for a period of 24 h. Before and after grazing, sward height, dry matter availability and nutritive value were evaluated. Grazing preference was visually assessed every 5 min for a period of 2.5 h after the afternoon milking. During the 3-year period 20 grazing events were evaluated. A randomized complete block design, with eight cultivars and three replicates, was used. Diploid cultivars showed greater herbage mass accumulation than tetraploid cultivars (P ≤ 0.05). No significant differences were obtained in the annual average crude protein content. Nevertheless, tetraploid cultivars showed a greater D value than diploid cultivars, except during the third year when the difference was not statistically significant. Dairy cows grazed more time on tetraploid cultivars. Considering, additionally, the residual herbage mass after grazing and the percentage of pasture utilization, diploid cultivars were less intensively grazed, suggesting a lower consumption by the cows.