Veteran Netherlands midfielder Rafael van der Vaart will miss the World Cup after injuring a calf muscle in his right leg.
The Dutch football association said on Wednesday the injury means coach Louis van Gaal will not include the most experienced player in his provisional squad in the final 23-man roster for Brazil.
The Hamburg star with 109 international appearances is the second important midfielder to drop out of Louis van Gaal's squad after Kevin Strootman suffered a season-ending knee injury in March.
Brazil would have been the 31-year-old Van der Vaart's third World Cup. The creative midfielder is a product of Ajax's storied youth academy who went on to play for Hamburg, Real Madrid and Tottenham.
Hi absence means Van Gaal will likely use Wesley Sneijder as his creative midfield general in Brazil. Sneijder was one of four joint top scorers at the 2010 World Cup with five goals.
Three-time runner-up the Netherlands opens its World Cup campaign in Salvador on June 13 with a repeat of the 2010 final against Spain. Van Gaal's team also plays Australia and Chile in a tough Group B.