Jack Midson gave Blue Square Premier leaders Oxford an FA Cup scalp as they sent former Cup giant-killers Yeovil, now in League One, crashing out.
Midson, who was involved in Histon's memorable FA Cup run last season, lobbed goalkeeper Richard Martin from 15 yards after good work by Matt Green.
Yeovil came close when Terrell Forbes shinned a far-post effort wide and Shaun MacDonald had a shot blocked.
Jon Obika saw a long-range shot saved late on as Oxford just held out.
Yeovil Town manager Terry Skiverton told BBC Somerset:
"There were too many players that had an off day. Everyone's sat in the changing room very disappointed.
"We were just poor. I'm disappointed because maybe we could have had a bit of fun and drawn a couple of decent teams in the future.
"It's the old adage of how hard do you fight? Do you fight harder as the underdog or harder as the champion? We were the champions today and didn't fight hard enough."
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