Tuchel is required to only name 23 players for a matchday panel and Ezri Konsa and Djed Spence both made the cut after recovering from injury.
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Konsa had missed the 2-1 win against Croatia on Thursday due to a knee injury, which is being managed by the England medical staff, while Spence was absent due to muscular problems.
Their return is a major bonus for England with Arsenal star Konsa expected to start at the Estadio Metropolitano.
However, as per UEFA's requirements, Jarred Branthwaite, James Garner and Josh King all miss out in the Spanish capital, but will remain with the group.
England need to win in Madrid to claim top spot in the group, but both sides have already qualified for the 2027 quarter finals.