Pompey have been drawn away to fellow League One side Ipswich in the second round of the Papa Johns Trophy.
The tie will take place at Portman Road in the week commencing November 21.
The sides have never previously met in this competition, although the Tractor Boys narrowly came out on top with a 3-2 league victory earlier in the season.
Kieran McKenna's side – currently sat second in the third tier standings – topped Southern Group H to guarantee a home draw in the last 32.
They thrashed Northampton 6-0 and enjoyed a 2-0 success over Arsenal U21s, but lost 1-0 in their final match at Cambridge.
Pompey, meanwhile, ended as runners-up behind AFC Wimbledon in Southern Group B, as they look to claim the silverware they lifted in 2019.
Ipswich – whose squad includes Gassan Ahadme, Conor Chaplin and Marcus Harness – are looking to progress beyond this stage of the competition for just the second time.
| Papa Johns Trophy Round Two – Southern Section |
| Colchester United v Bristol Rovers |
| Stevenage v Arsenal U21 |
| AFC Wimbledon v Sutton United |
| Milton Keynes Dons v Newport County |
| Ipswich Town v POMPEY |
| Forest Green Rovers v Cheltenham Town |
| Plymouth Argyle v Charlton Athletic |
| Chelsea U21 v Peterborough United |
| Papa Johns Trophy Round Two – Northern Section |
| Bolton Wanders v Barrow |
| Everton U21 v Mansfield Town |
| Salford City v Bradford City |
| Port Vale v Barnsley |
| Manchester United U21 v Wolverhampton Wanderers U21 |
| Grimsby Town v Accrington Stanley |
| Burton Albion v Tranmere Rovers |
| Lincoln City v Morecambe |