Southgate’s side take on the Croats in the first leg at Wolves’ Molineux home next Friday before travelling to Vinkovci for the second leg on Tuesday 14 October.
England U21s v Croatia U21s
UEFA Euro U21 Championship
Play-off first leg
5.45pm, Friday 10 October
Molineux, Wolverhampton Wanderers FC
Live on BT Sport
Call 0871 222 1877 for tickets
And the squad sees a return to the Under-21s group for Manchester United defender Luke Shaw, who was a regular in Southgate’s team in the early stages of qualifying before joining Roy Hodgson’s senior squad earlier this year.
Arsenal defender and former England U19s captain Calum Chambers receives his first Under-21s call-up, having made his senior debut last month.
Chelsea midfielder Lewis Baker is another player to join Southgate's squad for the first time, after captaining the U20s to a 6-0 win over Romania last month.
There is also a recall for Chelsea striker Patrick Bamford, who has been in impressive form on loan at Middlesbrough this season.
Southgate is without squad regulars James Ward-Prowse, Nathaniel Chalobah and Danny Ings, who are all ruled out through injury.
The squad will meet at St. George's Park on Sunday evening to begin their preparations for the first leg five days later.
Tickets are on sale for the Croatia game at Molineux now, priced at just £10 and £5 via the Wolves Ticketline on 0871 222 1877 or online here.
The Squad
Goalkeepers
Jonathan Bond (Watford), Jack Butland (Stoke City), Sam Johnstone (Manchester United).
Defenders
Calum Chambers (Arsenal), Eric Dier (Tottenham Hotspur), Luke Garbutt (Everton), Ben Gibson (Middlesbrough), Carl Jenkinson (West Ham United, on loan from Arsenal), Michael Keane (Burnley, on loan from Manchester United), Jamaal Lascelles (Nottingham Forest, on loan from Newcastle United), Liam Moore (Leicester City), Luke Shaw (Manchester United).
Midfielders
Lewis Baker (Chelsea), Tom Carroll (Swansea City, on loan from Tottenham Hotspur), Jake Forster-Caskey (Brighton & Hove Albion), Will Hughes (Derby County), Thomas Ince (Hull City), Alex Pritchard (Brentford, on loan from Tottenham Hotspur), Nathan Redmond (Norwich City).
Forwards
Patrick Bamford (Middlesbrough, on loan from Chelsea), Saido Berahino (West Bromwich Albion), Harry Kane (Tottenham Hotspur), Cauley Woodrow (Fulham).
