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Perennial ryegrass (PRG) is an important forage grown on dairy farms in temperate regions globally, including south-eastern Australia. A forage value index (FVI) providing information on the seasonal production of commercially available PRG cultivars is currently available. Despite the importance of the nutritive value of pasture in dairy farm systems, the nutritive characteristics of PRG cultivars are not currently included in the FVI as they are not routinely measured in cultivar evaluation trials. This study investigated differences between cultivar functional groups (diploid and tetraploid). It also examined differences between individual cultivars within seasons at four locations in south-eastern Australia and examined how trial location affects cultivar ranking. Samples were collected from existing cultivar evaluation trials over a 3-year period and analysed for nutritive characteristics. There were differences (p < 0.05) between diploids and tetraploids for metabolisable energy (ME) and neutral detergent fibre (NDF) in each season at each location with a few exceptions in summer and autumn. Crude protein (CP) differed between functional groups in some seasons at some sites. Spearman rank correlations within season were strong for ME between trial locations (r = 0.78–0.96), moderate to high for NDF (0.51–0.86) and variable for CP (−0.69–0.56). These findings provide guidance on methods for implementing nutritive value testing in cultivar evaluation trials and support the imminent inclusion of ME in the Australian FVI. The ranking of cultivars for ME was more consistent across trial sites compared to NDF and CP, suggesting the latter two traits, in particular CP, are more sensitive to environmental influences. Based on these results, we do not recommend the inclusion of CP as an individual trait in the Australian FVI. A significantly larger dataset and further research on the genotype by environment interactions would be needed to reconsider this. The addition of ME in the Australian FVI will lead to better cultivar choices by farmers and could lead to more targeted perennial ryegrass breeding programs. [ABSTRACT FROM AUTHOR]