Perennial ryegrass (Lolium perenne) is a perennial crop used in temperate regions as forage. In L. perenne breeding programs, persistency is an important trait. Poor persistency results in sward degradation and associated yield and nutritive value losses. Breeders assess persistency of accessions using visual scoring in field plots during the 2nd or 3rd growing season. This evaluation system is easy and cheap but is not free from human bias. In this study, the correlation between the scoring done by different breeders was only 0.243. As an alternative we have developed a methodology to assess persistency of L. perenne breeding materials based on vegetation indices (VIs) derived from Unmanned Aerial Vehicle (UAV) imagery. The VIs Excess green (ExG2), Green Leaf Index and Normalized green intensity (GCC) were found to provide consistent results for flights carried out under different light conditions and were validated by ground reference information. The correlation between the VIs and the percentage of ground cover extracted from on-ground imagery was 0.885. To test the implementation of the method we compared the ExG2 value based approach to selection with a visual score based selection methodology as applied by two breeders. The breeding decisions of Breeder A agreed well with decisions based on ExG2 values (74.6%), but those of Breeder B displayed a lower agreement (54.0%). In contrast, agreement between decisions based on different flights was very high (91.6%). The methodology was validated for general applicability. In summary, the results demonstrate that basing persistency selection in L. perenne breeding programs on ExG2 values from UAV imagery is likely to be more objective in comparison to the currently-used visual scoring method.