Plants especially need to adapt to all different light environments (shade, high light, etc.) due to the essential role of light in plant life. Either shade or high-light microenvironmental conditions are common for cool-season turfgrasses, such as perennial ryegrass (Lolium perenne L.). In order to study how a plant highly conserves microRNA, miR164-affected perennial ryegrass were studied under different light intensities. OsmiR164a-overexpression (OE164), target mimicry OsmiR164a (MIM164), and CRES-T (chimeric repressor gene-silencing technology) OsNAC60 (NAC60) transgenic plants and wild-type (WT) plants were evaluated in both field (shade and full sun) and growth chamber conditions (low, medium, and high PAR at 100, 400, and 1200 µmol s−1 m−2). Morphological and physiological analysis showed miR164 could fine-tune perennial ryegrass adaptation to changing light intensity, possibly via the regulation of target genes, such as NAC60. Overall, OE164 and NAC60 plants were similar to each other and more sensitive to high light, particularly under the field condition, demonstrated by smaller size and much poorer grass quality; MIM164 performed more like WT plants than either the OE164 or NAC60 plants. This study indicates the potential of genetic manipulation of miR164 and/or its targeted genes for turfgrass adaptation to changing light environments, and future research to further investigate the molecular mechanism beneath would be warranted.