Another win for either side would probably prove enough to secure progress, after both won their opener 3-2 last month.
Match preview
Having lost last season's playoff final, Bolton endured a slow start to the 2024-25 season, taking just four points from their first six league fixtures - but recent results have helped them inch up the League One standings.
Last Tuesday's 4-2 win over Northampton Town was Wanderers' third straight success, though it took a second-half turnaround to keep their unbeaten run going at the weekend.
Ian Evatt's side dragged themselves level after going two goals down, earning a point against struggling Shrewsbury Town, with Kyle Dempsey and Szabolcs Schon scoring to make it 2-2; however, they could not find a decisive third goal.
After the final whistle, an enraged Evatt was shown a red card by the referee, but he will hopefully have calmed down before leading his side out at the Toughsheet Community Stadium on Tuesday.
His team occupy 13th place in the League One table - just two points shy of the playoff places - as they turn their attention back to a EFL Trophy campaign they kicked off with a 3-2 win over Barrow in early September.
While Bolton edged a five-goal contest in their opening fixture, Aston Villa Under-21s managed the same feat against Fleetwood Town.
Completing his road to recovery from a long-term injury, Emi Buendia scored twice on his first competitive start since May 2023, volleying home from a Ross Barkley corner before tapping in a second towards the end of the first half.
Though the Villans then allowed Fleetwood to pull two goals back, substitute Kyrie Pierre produced a winner with three minutes remaining, so they sit alongside Bolton at the top of Group D.
This time around, not only will Josep Gombau's Young Lions aim to effectively wrap up qualification for the knockout phase, but also give more valuable minutes to two more senior stars slowly making their way back to full fitness.
Alongside Villa's youth prospects, there will be an opportunity for Tyrone Mings and Boubacar Kamara to take another step closer to long-awaited first-team returns.
Saturday's 3-2 defeat to Newcastle United in Premier League 2 saw the pair - sidelined for 14 months and eight months respectively - both complete the first half at Bodymoor Heath, as Villa slipped down to 17th place in the table.
Bolton Wanderers EFL Trophy form:
W
Bolton Wanderers form (all competitions):
L
W
L
W
W
D
Aston Villa Under-21s EFL Trophy form:
W
Aston Villa Under-21s form (all competitions):
D
L
W
W
L
L
Team News
Ian Evatt is set to rotate his Bolton squad in midweek, following a hectic schedule over the past few weeks, so John McAtee and Scott Arfield could come into the starting lineup.
While Chris Forino will be out of action for several months after surgery on a hamstring injury suffered against Arsenal in the EFL Cup, Carlos Mendes Gomes is now back in full training and could be offered some minutes on Tuesday evening.
Wanderers were forced to give a first league start to goalkeeper Luke Southwood after Nathan Baxter picked up an injury before Saturday's game, so it remains to be seen who will wear the gloves.
Meanwhile, Aston Villa duo Tyrone Mings and Boubacar Kamara should start in defence and midfield respectively, bolstering the visitors' young side with plenty of experience.
Top scorer Ben Broggio and Kobei Moore - son of former Villa player Stefan Moore - could feature in the final third, joining Jamaldeen Jimoh, who was signed from local rivals West Bromwich Albion last year.
Bolton Wanderers possible starting lineup:
Southwood; Toal, Santos, Johnston; Dacres-Cogley, Matete, Sheehan, Arfield, Schon; Collins, McAtee
Aston Villa Under-21s possible starting lineup:
Proctor; Rowe, Mings, Alcock, Patterson; Broggio, Amundsen-Day, Kamara, Jimoh-Aloba; Moore, Taylor
We say: Bolton Wanderers 1-1 Aston Villa Under-21s
The points could be shared in Bolton, as the home side may be fatigued from negotiating a frantic fixture list since midway through September, allowing Villa's young guns to secure a draw.