A 1-0 defeat to League Two (fourth tier) rivals Mansfield Town condemned Rovers to relegation on goal difference after fellow strugglers Wycombe Wanderers beat already demoted Torquay United 3-0.
With both sides tied on 50 points from 46 matches, Rovers were down and out for the first time since they joined the League in 1920.
"I think this is the saddest day in the history of the club," said chairman Nick Higgs. "We have reached rock bottom and now we have to bounce back."
Rovers fans invaded the pitch after the final whistle, chanting 'sack the board' before mounted police and stewards cleared the field.