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The match ended level in normal time, with Marseille taking the lead twice. England forward Mason Greenwood opened the scoring from the penalty spot, before Brazil’s Igor Paixão added another goal in the 56th minute.
Toulouse responded on both occasions through Yann Gboho in the 13th minute and England defender Charlie Cresswell in the 60th minute.
The tie was ultimately decided in a penalty shoot-out that favoured Toulouse FC. The visitors missed one attempt through Venezuela’s Cristian Cásseres Jr., while Marseille failed with two efforts from their Argentine captain Leonardo Balerdi and England’s Ethan Nwaneri.
The defeat marks the second competition Marseille have exited this season, after failing to reach the play-off round for the UEFA Champions League. The disappointment led to the dismissal of Italian coach Roberto De Zerbi, who was replaced by Senegal’s Habib Beye.