Barnsley beat Duncan Ferguson's side on Saturday, while Fleetwood secured a 5-2 triumph over Accrington Stanley.
Match preview
A fourth loss on the trot for Rovers confirmed their immediate relegation to League Two after only gaining promotion to League One last year.
Ferguson's men conceded three first-half goals against Barnsley in the eventual 5-1 thrashing, their fourth reverse in succession.
A 1-0 win over promotion-chasing Sheffield Wednesday in late March ended a three-month drought without success, but that triumph was not the fillip Forest Green hoped for.
The lowest-ranked side in the division has conceded 81 times in 42 games, the worst defensive return in the competition, and their inability to score consistently — 30 goals is also the lowest in the league — meant they had little chance of beating the drop.
After instant relegation from League One, all Rovers can do is regroup in the fourth tier and strive for immediate promotion.
Tuesday's visitors have little to play for, with Fleetwood 13 points clear of the bottom four and 17 points adrift of a playoff spot.
The 13th-placed side did return to winning ways this weekend, scoring five times in a 5-2 win over Accrington after a three-match run without success.
Scott Brown's team picked up one point from an available nine, losing twice in that period to Cambridge United and Bristol Rovers.
They have one of the league's best road records, with only six teams accruing more than their 28 points away from home, and they might have been in the playoff conversation had their record at Highbury Stadium not been the fifth-worst in the competition.
Both sides played out a 1-1 draw last October in the reverse fixture in Lancashire.
Forest Green Rovers League One form:
L
W
L
L
L
L
Fleetwood Town League One form:
W
W
D
L
L
W
Team News
Tyler Onyango has not played for Forest Green since going off injured against Morecambe, and the central midfielder is still out with a hamstring injury.
Jahmari Clarke tore a tendon soon after his temporary move to Rovers and has not featured since February.
Kyle McAllister was withdrawn after 23 minutes against Barnsley, making an appearance unlikely on Tuesday.
Fleetwood have been without Harvey Macadam since the wide attacker went off after 15 minutes against Bristol Rovers.
Jack Marriott netted a hat-trick in Saturday's triumph over Accrington, and the striker should keep his place in the side against Rovers.
Forest Green Rovers possible starting lineup:
Doohan; Bernard, Casey, Cooper, Robson; Peart-Harris, McGeouch, Bunker, Garrick; Savage, Omotoye
Fleetwood Town possible starting lineup:
Lynch; Rooney, Sarpong-Wiredo, Earl, Johnston; Vela, Warrington; Omochere, Stockley, Patterson; Marriott
We say: Forest Green Rovers 1-2 Fleetwood Town
Rovers have scored one goal in four of their last five games at the New Lawn, but they are expected to be beaten by one of the league's best travelling sides.