Methiozolin is a new isoxazoline herbicide that has scarcely been tested in Kentucky bluegrass turf. A field trial was conducted in Blacksburg, VA, to determine response of 110 Kentucky bluegrass varieties and winter annual weeds to sequential fall applications of methiozolin. At 1.5 and 6 mo after initial treatment (MAIT), Kentucky bluegrass injury I30 values (predicted methiozolin rate that causes 30% Kentucky bluegrass injury) ranged between 3.4 to more than 10 times the recommended methiozolin rate for annual bluegrass control. Methiozolin at all rates reduced cover of annual bluegrass, common chickweed, corn speedwell, hairy bittercress, mouseear chickweed, and Persian speedwell but increased cover of parsley-piert. For all varieties, methiozolin at 2 kg ai ha−1 increased Kentucky bluegrass cover, turf quality, and turf normalized difference vegetation index (NDVI) relative to the nontreated check at 6 MAIT. Kentucky bluegrass relative cover change (RCC) was attributed primarily to weed control but was inversely correlated with methiozolin rates because of increased weed control and reduced Kentucky bluegrass growth. Despite the decline in RCC with increasing methiozolin rates, most Kentucky bluegrass varieties treated with the highest methiozolin rate (6 kg ha−1) still had greater Kentucky bluegrass cover than the nontreated check at 6 MAIT. Results from this study indicate that two fall applications of methiozolin at rates beyond that previously reported for annual bluegrass control can safely be applied to a broad range of Kentucky bluegrass varieties spanning most of the known genetic classifications.