Southampton's youngsters will open their group stage Checkatrade Trophy campaign against League Two side Colchester United in early September.
Radhi Jaïdi side will then travel to Cambridge United in October, before concluding their Group B fixtures against League One outfit Southend United in mid November.
All ties will take place away from home, with Saints making their third successive appearance in the competition.
Colchester United vs Saints
Tuesday 4th September, Colchester Community Stadium, 7:45pm
Cambridge United vs Saints
Tuesday 9th October, Abbey Stadium, 7:45pm
Southend United vs Saints
Wednesday 14th November, Roots Hall, 7:45pm