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The defeat means that Real Madrid have now little chance of finishing top of Group H, and as a result they will probably face a tough rival in the last 16 of the competition.
Madrid's defeat, coupled with Zenit St Petersburg's win against Bate Borisov, means failure to beat Bate in Ukraine on Nov. 29 or a defeat against the Zenit on Dec. 10 could see Bernd Schuster's side knocked out of the competition altogether.
Alessandro Del Piero was once again Real Madrid's main tormentor. The Juventus striker had scored the first goal in Juventus' 2-1 win over Madrid two weeks ago and he opened the scoring after just 17 minutes in the Santiago Bernabeu stadium, scoring with his left foot from inside the penalty area.
Madrid had started the game with Ruud van Nistelrooy back in their starting 11 but without central defender Pepe, who had failed a late fitness test.
Royston Drenthe also started for Madrid with Arjen Robben on the substitutes' bench.
The home side started brightly but was unable to create any clear-cut chances against the Italians who were, as usual, well organized in defense.
Madrid had most of the possession after the goal, but was unable to break down a side that was perfectly positioned and hardworking.
It is over 45 years since Juventus last won in the Santiago Bernabeu, but the Italians rarely looked troubled, even as Real Madrid stepped up the pace after half time.
Juventus doubled their lead after 67 minutes after Del Piero scored a direct free kick. That virtually spelt the end of the home side's resistance and although Schuster even brought on Javier Saviola for Van Nistelrooy, there was nothing his team could do to get back on level terms.