Field studies were conducted on southern highbush blueberry in Elizabethtown and Rocky Point, NC, in 2019, 2020, and 2021 to determine tolerance to 2,4-D choline as a postemergence-directed application. In separate trials for younger and older bearing blueberry bushes, both 2,4-D choline rates and application timing were evaluated. Treatments included 2,4-D choline at 0, 0.53, 1.06, 1.60, and 2.13 kg ae ha–1 applied alone in winter during dormancy, and sequential treatments at 0.53 kg ae ha–1 followed by (fb) 0.53, 1.06 fb 1.06, 1.6 fb 1.6, or 2.13 fb 2.13 kg ae ha–1. The first application of the sequential treatments was applied in winter followed by another application in spring during early green fruit. Injury to blueberry from 2,4-D choline treatments was not observed for either maturity stage, and fruit yield was not affected by any of the treatments. Differences among treatments were not observed for fruit soluble solid content (SSC) in older bushes, or for fruit pH, SSC, and titratable acidity (TA) in younger bushes. In older bushes, fruit pH and TA had rate-by-timing interactions, and TA had a farm-year interaction with differences at Rocky Point in 2019 and Elizabethtown in 2020, but biologically no pattern was observed from the treatments.