The two teams will be joined by Exeter City and Chelsea Under-21s in the Southern Group E, with the top two progressing to the knockouts.
Bristol Rovers have not adapted to life in League Two in the manner that manager Joey Barton would have hoped.
The Gas were relegated from League One last season - after a five-year spell in the third tier - finishing dead last on 38 points, a run of five consecutive losses to finish the season cementing their relegation.
The owners backed Barton as the man to turn their fortunes around but things have not worked out that way so far, with the club sitting 22nd in the table and having collected just one win in their first five matches.
More is surely expected of a side coming down from League One, but Rovers cannot seem to find form and were held to a frustrating 1-1 draw by 10-man Barrow at Holker Street on Saturday, throwing away a lead and failing to capitalise on their near hour-long man advantage.
Perhaps Barton can find more joy in the cup competitions, but they will open their EFL Trophy campaign against a team who last season topped the league they are currently struggling in.
Cheltenham are holding their own since their promotion to League One, despite experiencing a fairly hectic start to the campaign.
They earned a 2-1 victory against pre-season title favourites Ipswich Town in their third game but also hold the second-worst defensive record in the league, after conceding in every game thus far and letting in three on two occasions.
The Robins have also scored in every game, however, and manager Michael Duff will certainly have seen enough from his side to give him hope that they will retain their place in the third tier this campaign.
Callum Wright has been a huge hit during his second spell on loan from Leicester City, already contributing three goals and an assist from the centre of midfield, including the goal in their most recent game - a 1-1 draw away at promotion-chasing Burton Albion.
Rovers made three changes for their game against Barrow, with Sam Nicholson, Connor Taylor and Brett Pitman replacing Jack Baldwin, Paul Coutts and Harvey Saunders, but whether they did enough to retain their place in the starting XI is up for debate.
Duff is known to mix up his teams for club competitions, so he may go with a lineup more closely resembling the one against Gillingham in the League Cup than the most recent XI.
That would see George Lloyd and Taylor Perry return to the team, amongst others, but - considering their opponent's recent form - the manager is more likely to stick with his more attacking formation of a 3-5-2.
Bristol Rovers possible starting lineup:
Belshaw; Hughes, Anderton, Taylor; Grant, Finley, Anderson, Rodman; Collins, Nicholson; Pitman
Cheltenham Town possible starting lineup:
Evans; Long, Freestone, Boyle; Blair, Perry, Chapman, Sercombe, Hussey; May, Lloyd
We say: Bristol Rovers 1-2 Cheltenham Town
The possibility of Cheltenham keeping up their run of scoring and conceding in their recent games seems a strong one, but we fancy the away side to add to Barton and his team's troubles.
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