Anthocyanins are increasingly being used as natural alternatives in medicinal, food, and industrial products. However, production of anthocyanin extract is often inefficient due to agronomic limitations. On the other hand, the use of turfgrasses for anthocyanin production has been suggested to increase yield twofold. Rough bluegrass (Poa trivialis L.) cultivar ‘Havana’ has been shown to increase anthocyanin content by 117-fold under high light treatment, exhibiting concentrations similar to current anthocyanin sources, and could be an alternative source of anthocyanin. The objectives of this research were to evaluate variation in anthocyanin content in 20 accessions and five cultivars of rough bluegrass treated with blue light and to evaluate phenotypes associated with anthocyanin content in this species to help develop germplasm screening tools. Following blue light treatment, rough bluegrass US cultivars ‘Laser’, ‘ProAm’, ‘Sabre’, ‘Colt’, Havana, and accessions originating from Germany produced statistically greater quantities of anthocyanin compared with other accessions. Phenotypes associated with anthocyanins, including total phenolic content, total flavonoid content, specific leaf area, leaf cuticular wax, and the chlorophyll:carotenoid ratio, were variable for all accessions and cultivars. However, accessions producing greater quantities of anthocyanin exhibited less cuticular wax. Results of a principle component analysis showed that there was a significant negative correlation between leaf cuticular wax and anthocyanin content. These data demonstrate that phenotypes associated with increased anthocyanin content, especially leaf cuticular waxes, could serve as screening tools for evaluating other turfgrasses as anthocyanin sources and may therefore help to maximize industrial anthocyanin production.