Huddersfield remain at the summit, Norwich City move into second after Newcastle's slip up at home to Wolves.
Scunthorpe move into top spot in Sky Bet League One after Bolton Wanderers' defeat to 10-man Walsall. Port Vale maintained their 100% home record.
Plymouth's Devon derby win made it six successive wins to remain top. Doncaster, Crewe, and Morecambe all won to keep pressure on the leaders.
The highlights of a thrilling Round Two saw Accrington, Northampton and Gillingham beat top-flight opposition in Burnley, West Brom and Watford.
With a new format for Season 2016/17, 64 teams will be aiming to follow in Barnsley’s footsteps from 2015 and be victorious at Wembley in April.
Get our newsletter