Rovers Under-23s' 11-game unbeaten run finally came to an end at the hands of Brighton & Hove Albion at the Lancashire FA HQ on Friday evening.
Two first half goals sealed the team's fate, as first Viktor Gyokeres and then James Tilley did the damage on a chilly night in Leyland.
Rovers had their chances, but couldn't find a way past a resolute Albion defence, and failed to score for the first time in their last five matches in all competitions.
Damien Johnson included four players with first team experience in the XI, as Jayson Leutwiler, Willem Tomlinson, Lewis Travis and Joe Nuttall all started in Leyland.
Joe Rankin-Costello was out of the squad through injury, whilst visitors Brighton included Tom Dallison is the side, who spent the first half of this season on loan at our near neighbours Accrington Stanley.
A strong start from Rovers saw Willem Tomlinson test Robert Sanchez from range, but the visiting 'keeper beat away the midfielder's rasping effort.
But five minutes later and Brighton took the lead from their first effort as Gyokeres brilliantly bent the ball into the top right corner from 25 yards out.
And soon after the visitors had their second as Tilley beat Leutwiler excellently after connecting with a Henrik Bjordal cross from the left.
Rovers had to find a response and Tomlinson fired over the bar after Sanchez could only push away Jack Evans' cross from the left.
Dan Butterworth then blasted over from close range as Rovers began to probe the Albion goal with half time approaching.
With three minutes of the half remaining, Nuttall saw Sanchez just about do enough to tip his effort wide of the post.
After the break, Joe Grayson tested Sanchez from a free kick before John Buckley curled a 20-yard shot wide as Rovers continued to create chances but fail to find a route past Brighton's Spanish stopper.
Moments later, Buckley's cross was skewed wide by Nuttall, whilst at the other end, Tilley latched onto a Jordan Davies centre but Leutwiler was there to beat over the bar.
Leutwiler was again called into action a moment later as he tipped over an Owen Moore strike, with the Seagulls looking to kill off any chance of a possible Rovers comeback.
Time was running out, and with just over 20 minutes to go, Nuttall looped a header wide of the post, whilst the Seagulls threatened on the counter, but the impressive Tilley drilled well wide of Leutwiler's upright.
Rovers huffed and puffed, but couldn't find a way through. The visitors were professional and saw the job through. A positive for Johnson's side is that they still remain clear at the top of the Premier League 2 table.
The team have a break from Premier League 2 action, as next up they head to Cardiff in the Premier League Cup on Thursday 22nd February, kick-off 12pm.
Rovers Under-23s: Leutwiler, C. Doyle, Evans, Tomlinson, Magloire (Mansell, 57), Grayson, Travis, Hardcastle (c), Nuttall, Butterworth (Simmonds, 46), Buckley (Vale, 74).
Unused substitutes: Carter, Albinson.
Booked: Evans, 56
Brighton Under-23s: Sanchez (Billings, 81), Forbes, Cox, Davies, Moore, Dallison (c), Tilley (Kerr, 86), Alzate, Gyokeres, Connelly, Bjordal.
Unused substitutes: Mateju, Cochrane, Cashman.
Goal: Gyokeres, 15, Tilley, 22
Booked: Davies, 59, Dallison, 83