Meanwhile, the visitors are featuring in their first game of this year's competition, also tasked with encounters against Wrexham and Salford City.
Match preview
With a long-term contract and in charge of a team that were relegated last season, Darren Moore could have done with a strong start with Port Vale in 2024-25.
Instead, the Staffordshire outfit have accumulated just four points from as many league games, as well as losing their most recent two fixtures.
Furthermore, a total of seven goals have been shipped against Barrow and Doncaster Rovers respectively, Vale now down in 16th position in the League Two table.
Coincidentally, each of their victories this season have come versus the same opponents, with 2-0 triumphs being posted versus Salford City in the fourth tier and this competition respectively.
While Moore will not view this tournament as a priority, he will be eager to get his side back on track as soon as possible at a time when a win could book their place in the next round.
As for Wolves Under-21s, they will be attempting to give a better account of themselves than they did during last season's campaign.
Just three points were recorded versus Notts County, Lincoln City and Derby County respectively, conceding seven goals in the process.
James Collins will be disappointed with how Wolves have started in Premier League 2, earning just one point from meetings with Nottingham Forest, Arsenal and Brighton & Hove Albion.
The draw in question came in the most recent encounter at the young Seagulls on Saturday, extending their winless streak in that competition to seven matches stretching back to March 15.
Port Vale EFL Trophy form:
W
Port Vale form (all competitions):
W
L
D
W
L
L
Wolverhampton Wanderers Under-21s form (all competitions):
Team News
Moore could make similar alterations to how he lined up for the Salford game, the likes of Ben Amos and Jesse Debrah in line to feature in defensive areas for Vale.
George Byers and Ethan Chislett should play in midfield, with Jayden Stockley getting the nod down the middle of the attack.
Collins is likely to name many of the players who featured against Brighton on Saturday, yet there are possibilities for some high-profile names to earn minutes.
Each being 18 years of age, there is theoretically a chance that Pedro Lima and Bastien Meupiyou could be handed a place in the squad.
Port Vale possible starting lineup:
Amos; Debrah, Heneghan, Hall; Cover, Byers, Garrity, Hart; Chislett, Stockley, Curtis
Wolverhampton Wanderers Under-21s possible starting lineup:
Benjamin; Okoduwa, Pond, Voice, Ket-Diyawa; Rawlings, Whittingham; Esen, Edozie, Farmer; Angel
We say: Port Vale 2-1 Wolverhampton Wanderers Under-21s
With Vale likely to make some changes, there is the prospect of Wolves earning at least a share of the spoils. However, we can only back the more experienced home side to come through this contest, even if they are made to work hard for it.