Plymouth beat Tranmere 3-2 to condemn Rovers to relegation and end their 94-year stay in the Football League.
Argyle's win also all-but confirmed their place in the Sky Bet League Two end of season play-offs.
Plymouth's 20-goal top scorer Reuben Reid fired Argyle into a seventh-minute 1-0 lead from the penalty spot.
The spot-kick was awarded after Max Power felled Dominic Blizzard as the midfielder broke into the area.
Power atoned for his foul by restoring parity with a thumping strike in the 34th minute.