Both teams are currently in League One, and they played out a 1-1 draw when they met at Oakwell back in September.
Match preview
Port Vale were very impressive in the group stages, winning all three games without conceding a goal to achieve progression to the first knockout round.
To secure their place as group winners, Vale ran out 2-0 winners against Wolverhampton Wanderers Under-21s in their last EFL Trophy match, with Malvind Benning and Ademipo Odubeko scoring late goals.
In terms of their league performances, Darrell Clarke's side have made a very good start in their first season back in the third tier, currently 10th in the standings, just four points off the playoff places.
Last time out, the Valiants triumphed 1-0 over Charlton Athletic on home soil, a scoreline that they have made a habit out of winning by, having defeated Lincoln City and Cambridge United in the same fashion in their previous two victories.
Daniel Butterworth netted the only goal of the game against the Addicks just before the hour mark, with the backline then putting in a solid defensive display to wrap up all three points.
Barnsley were not quite as dominant as Vale in their EFL Trophy group, but managed to win two of their three games to book their place in the first knockout round.
The Tykes' fate came down to the final game, in which they comfortably dispatched of Doncaster Rovers 4-2, after racing into a 4-0 lead prior to half time, with Josh Martin and Fabio Jalo scoring braces.
Turning to the league, Michael Duff's side have made a solid, if not spectacular start to the campaign, having been relegated from the Championship last season, and they are sixth in the table as things stand.
Recent form has really started to pick up for Barnsley, winning their last four games in all competitions, also securing progression to the second round of the FA Cup with a 2-1 win away at Bolton Wanderers.
In their last match, the Tykes were 3-1 winners against MK Dons at Oakwell, while in the two League One games prior to that, they kept clean sheets, beating Shrewsbury Town 1-0 on their travels, and defeating Forest Green Rovers 2-0 on home soil.
Port Vale EFL Trophy form:
W
W
W
Port Vale form (all competitions):
L
W
D
L
L
W
Barnsley EFL Trophy form:
L
W
W
Barnsley form (all competitions):
L
L
W
W
W
W
Team News
Given that both sides are likely to want to prioritise their league campaigns, there should be plenty of changes, with some young players given opportunities.
For Port Vale, 20-year-old Ademipo Odubeko may come into the side, having had to settle for substitute appearances in recent matches, potentially replacing James Wilson in the frontline.
Sam Robinson could come into the defence, starting on the right in a 5-3-2 system, with the more experienced David Worrall dropping to the bench.
Aidan Stone is also likely to feature between the sticks, as he was the choice in the 2-0 victory against Wolves Under-21s, with the goalkeeper replacing Jack Stevens in the starting lineup.
Turning your attention to Barnsley, striker Slobodan Tedic is unlikely to be involved, as he did not feature in the 3-1 win against MK Dons on Saturday, after sustaining an ankle injury against Shrewsbury in the prior match.
James Norwood is also a doubt in attack, having struggled with an injury recently, so Fabio Jalo may be chosen to partner Jack Aitchison.
Youngsters Will Lancaster and Jasper Moon last featured for the Tykes in the EFL Trophy match against Doncaster, and they could be given more first-team game time on Wednesday night.
Port Vale possible starting lineup:
Stone; Smith, Jones, Hall; Robinson, Garrity, Conlon, Benning, Butterworth, Massey, Odubeko
Barnsley possible starting lineup:
Collins; Edwards, Lancaster, Moon; Williams, Kane, Phillips, Connell, Cadden; Jalo, Aitchison
We say: Port Vale 0-1 Barnsley
It was a closely-fought encounter the last time these two sides met, with nothing being able to split them on that day. However, Barnsley have been in good form recently, and should possess the quality to edge into the next round, by a one-goal margin.