Despite largely bossing the game, Rovers Under-23s were defeated for the first time in five games in all competitions against Derby County Under-23s at Leyland.
A Cameron Cresswell brace put the visitors in command before Jack Vale pulled one back on the hour.
But an expert free kick from Calum MacDonald put the game out of reach for Damien Johnson's side, who will count themselves unfortunate not to take something from the clash.
The visitors started the stronger at Leyland, with Sven Karic cracking the top of the crossbar direct from a free kick just seven minutes into the encounter.
At the other end, Josh Barnes' poor clearance only went as far as Matty Platt, who almost lobbed the Derby 'keeper from the halfway line; his effort dropping just wide of the right-hand post.
Rovers were growing into the game and good play involving Lewis Thompson and Vale saw the Welshman almost squeeze the ball past Barnes just after the half hour mark.
With six minutes to go until the break, the Rams worked the opening goal of the game through a devastating counter attack.
Kyle McAllister charged upfield and his cross from the left was swept into the top corner by Cresswell, giving Andy Fisher no chance.
Rovers responded well and a neat swivel and turn from Hayden Carter was held by Barnes just a moment later.
Carter went close again just a minute later as he slammed a strike wide after a through ball from John Buckley found him in space.
Rovers will have counted themselves unfortunate to go in a goal down, with their play deserving more than the scoreline showed.
The hosts were quickest out of the traps at the start of the second half, and Vale forced Barnes into a save with his legs just seconds after the interval, before Thompson smashed a shot wide of the target.
But Johnson's men were dealt a sucker punch soon after, as Aaron Eyoma's cross was emphatically headed home by Cresswell.
However, Rovers forced a way back into the encounter when Thompson's pinpoint cross found Vale at the back post, who headed down and into the back of the net just before the hour.
For all Rovers' probing, it was Derby who grabbed the next goal as MacDonald sensationally curled home a free kick into the top corner from 20 yards.
It was a bitter blow for Rovers, with the goal knocking the stuffing out of the group, who knew a comeback would be unlikley with only 13 minutes remaining.
Fisher was called into action in the dying embers of the game, most notably saving from McAllister as the night ended in disappointment for Johnson's side.
Rovers: Fisher, Platt, Thompson, C. Doyle (Rankin-Costello, 58), Magloire, Grayson (c), Carter (Simmonds, 71), Tomlinson, Vale, Mols, Buckley.
Unused substitutes: Winterbottom, Paton, Saadi.
Goals: Vale, 58.
Booked: Magloire, 76, Grayson, 90.
Derby County: Barnes, Gordon (Rashid, 62), Karic, Brown, Hunt (c), MacDonald, Mitchell-Lawson, Eyoma (Shonibare, 75), Cresswell, Babos, McAllister.
Unused substitutes: Greco, Yates, Al-Hussaini, Shonibare.
Goals: Cresswell, 39, 51, MacDonald, 77.
Booked: Cresswell, 90.
Referee: Simon Mather