The only goal of the game came just before-half-time when Ollie Palmer kept his cool to roundBrad Wade and score,
Guiseley’s joint-manager Marcus Bignot said: “Obviously we are really proud of the group, to play the way we did against a team a few divisions above.
“On another day we could have been in the hat for the second round, it just wasn’t meant to be in the end.”
Non-league Guiseley welcomed back the majority of their first-team squad after the disruption caused a week ago through suspensions, injuries and Covid protocols.
The Dons started strongly with Palmer forcing Wade into a smart stop to keep the scores level early on.
The Lions started to settle into the match and had an opportunity of their own when Jamie Spencer hit a shot on goal confidently and forced Dons goalkeeper Nik Tzanev into making the save.
The hosts would get the opener just before 45 minutes when Palmer broke the deadlock
At the beginning of the second-half there was a key chance for The Lions, the hero of the last round Lebrun Mbeka was found behind the home defence, his effort though on the half-volley went over the bar.
Guiseley would keep trying to get the leveller, with a corner causing the home side problems, a scramble ensued but no shot on goal came from it and eventually the danger was cleared.
AFC Wimbledon starting 11: Tzanev, Guinness-Walker,, Csoka, Woodyard, Alexander (Lawrence 80), Hartigan, Assal, Rudoni, Palmer (Pressley 65), McCormick (Chislett 80), Heneghan. Unused substitutes: Mebude, Oualah, Charles, Jenkins, Biler, Bartley
Guiseley starting 11: Wade, Cantrill, Nicholson, Ekpolo, Hull, Bencherif (Hollins 69), Hey (Gratton 52), Mbeka, Spencer, Thewlis (Tuton 63), Hutchinson. Unused substitutes: Brown, Lambton, Metcalfe, Stones, Thompson.
Referee: M Woods (England).