The former Tranmere defender who missed much of last season due to injury was on the road to recovery from medial ligament damage sustained towards the end of last season until pulling his hamstring this afternoon.
Speaking to the Official Club website, reserve team coach Steve McCall said : "It's a blow for John, who was looking very good in the game and closing in on returning to full fitness.
"He has been moving forward with every game but this is disappointing for him. Hopefully it's nothing too serious but it's a step back for him unfortunately."
The match which kicked off at 1pm at the Playford Road Training Centre finished 0-0.
Town Side : Peat, Hogg (Flack 78), Okay, Murray (Reid 78), McGreal (Krause 53), Collins (Synott 46), Abidallah (Barron 61), France (Kamara 61), Abidallah 76), Armstrong, Westlake (Atay 46).