The former Dallas Cowboys quarterback shot 9-over 79 in local qualifying in Texas, finishing 11 shots outside the cutoff.
Romo, now a broadcaster, must wait at least another year for another chance and has not reached the final qualifying stage since 2010.
Other former athletes, including Aaron Hicks, Tyler Clippard, Mardy Fish and J.T. Miller, remain in qualifying for the June 15-18 tournament at Shinnecock Hills.
Will the PGA Tour welcome back its former stars, or will they face harsh penalties for joining the now-faltering LIV league?
After losing billions, is the Saudi PIF's exit a failed sportswashing attempt or a strategic pivot for professional golf's future?