Holland winger Ryan Babel will miss Euro 2008 after tearing ankle ligaments in training.
Babel is expected to be out of action for several weeks and will therefore not take part in the summer showpiece in Austria and Switzerland.
The Liverpool attacker sustained the injury a day before Holland play their final pre-tournament friendly, against Wales in Rotterdam.
Holland coach Marco van Basten has not named a replacement for the 21-year-old although he is permitted to do so once Babel has been examined by a UEFA doctor.
The Dutch will line up alongside France, Italy and Romania in Group C. Their first match is on June 9 against world champions Italy.
Despite the injury blow, Holland boss Marco van Basten has delivered an ominous warning for John Toshack's Wales ahead of their meeting at Feyenoord's sold-out De Kuip stadium, demanding a flurry of goals to send his side into the Euro 2008 finals on a high.
That will be worrying for Welsh ears, because the last time they faced Holland they were thrashed 7-1 in Eindhoven in 1996, and Van Basten clearly wants a similar scoreline against a country they have beaten on all five previous meetings between the side.
Van Basten was disappointed with Thursday's 1-1 home draw against Denmark, when he fielded a side much closer to the one expected to face Italy in their opening group C match on June 9 in Berne.
"We must be more clinical and take our chances. This team has matured over the last two years and everywhere the expectation is so much higher, not surprisingly," he said.
"But we cannot allow ourselves to let teams off the hook when we are making chances and on top."
Former Manchester United ace Ruud van Nistelrooy scored early on against the Danes, but Holland could not build on that opener.
And now Wales could feel the backlash with a young, inexperienced side set to line up against the Dutch.
Van Basten said: "It is a bad thing for us, we are not cashing in on the numerous chances we create. This is often a Dutch problem, and one we must solve before we go to the finals."
The former Dutch master, though, was happy with Van Nistelrooy's contribution, the Real Madrid man only joining up with the squad on Monday after being excused the early training work along with other stars who had been involved in big matches of late.
Manchester United's Edwin van der Sar - the Red Devils' spot-kick hero in the Champions League Final - was another later arrival in camp.
"Van Nistelrooy took his chance well and looked sharp. But others must start to score as well," added the coach.
Before Babel broke down Van Basten reduced his squad to 23 after Wigan defender Mario Melchiot proved his fitness after struggling with a groin problem. The three unlucky players to step down were keeper Sander Boschker, Denny Landzaat and Khalid Boulahrouz.
Wales could find themselves facing virtually Van Basten's strongest side, but Arsenal's Robin van Persie is unlikely to be risked as he is still suffering from a thigh problem.
Van Basten, perhaps significantly, made a point that the forward had been risked too soon by the Gunners at the tale end of the season.
He said: "We are not going to do that because Robin is a special player we would rather see in the side to face Italy.
"He came back too quickly last season and had a relapse. Our doctors believe it is better to allow him to recuperate as long as possible rather than risk him being injured in a friendly."
Aston Villa full-back Wilfred Bouma will get his chance to clinch a place in the starting line-up against the Italians, as will Real Madrid's former Chelsea man Arjen Robben, who was replaced late on by Babel against the Danes.The KNVB said a replacement for Babel was being discussed.
The Netherlands are in Group C along with France, Italy and Romania for the June 7-29 tournament in Austria and Switzerland.