Gosport crashed out of the Russell-Cotes Cup at Privett Park last night.
Alex Pike’s men were beaten 1-0 by Winchester City, with Gary Fulker netting the deciding goal from the spot.
Borough skipper Dave Birmingham had seconds earlier missed his own penalty kick and that mistake proved costly.
With important Southern League matches coming up, Gosport boss Pike rested several of his senior players.
So too did Winchester manager Guy Butters, as his side currently top the Wessex League premier division table.
And it was the visitors who started best and better finishing by Zak Glasspool and Mike Dixon may well have given them the lead.
But on 10 minutes Borough were awarded a penalty by an eagle-eyed referee who noticed a handball in the area.
Up stepped skipper Birmingham only to see his well-struck penalty beaten away by City keeper Rory Anderson.
The youthful visitors then made Borough pay.
Play broke to the other end and Gosport defender Dan Mobsby handled the ball to prevent it reaching Dixon.
Up stepped Fulker to thump the spot-kick past keeper Mark Brown and give his side the lead.
The goal lifted Winchester to a higher plane and only woeful finishing by Dixon spared Borough further embarrassment.
In a rare foray into the Winchester half, Borough were inches away from an equaliser as striker Liam Sewell – on loan from the Hawks – cut a path through only to see his fierce shot cannon back into play off a post.
As play switched back in favour of Winchester, substitute striker Suhael Odeh met a cross at the far post to slam his finish against the bar.
Late on – and with the match virtually won – Winchester turned up the heat once more.
And it took a goalline clearance from Pete Castle to prevent Tom Dunsford from doubling the visitors’ advantage.
But in the final analysis one goal was more than enough to send Borough crashing out of the competition and gift Winchester a semi-final tie against Bemerton Heath Harlequins.