Oxford City bowed out of this year's competition at the second hurdle after a disappointing display in Hampshire.
In a game made difficult by a strong wind, it was the visitors who started the stronger, with their neat approach play penning back the home side.
Midway through the first half, City had a great chance of taking the lead when Winchester defender Mark Zawadski made an almighty hash in attempting to clear, but Anthony Alleyne could only find the side-netting, with the goal gaping.
Winchester had a couple of near misses, but nothing on target as the first-half ended goalless.
Much was expected of the visitors in the second period, but it was not to be as Winchester hit the winner on 48 minutes.
Jamie Musselwhite got the better of Adam Durrant to lob a wind-assisted effort over Lee Farrow, who had replaced the injured Carl Wilkins in goal.
The nearest City came to getting an equaliser came when Alleyne's header rebounded off the keeper, but after that, they went out with a whimper.
Oxford City: Farrow, Durrant, G Williams, Avery, Craker (Lane 50), Takriti, Burnley, Merritt, Alleyne, Keen (Stewart 70), Jones (R Williams 28).
Comments: Our rules
We want our comments to be a lively and valuable part of our community - a place where readers can debate and engage with the most important local issues. The ability to comment on our stories is a privilege, not a right, however, and that privilege may be withdrawn if it is abused or misused.
Please report any comments that break our rules.
Read the rules hereComments are closed on this article