Gareth Bale has been ruled out of Wales' World Cup Qualifiers against Georgia and the Republic of Ireland through injury.
The forward had been a major doubt after missing Real Madrid's 2-0 win over Espanyol due to the calf muscle strain he suffered against Borussia Dortmund in the Champions League last Tuesday.
And, following a scan on the injured calf, Bale will definitely not be available for the crucial fixtures, dealing a major blow to Wales' hopes of qualification.
Bale was ruled out following talks between the Football Association of Wales and his club Real Madrid, and Barnsley striker Tom Bradshaw has been called up to replace the 28-year-old.
An FAW statement read: "The Football Association of Wales can confirm that Gareth Bale will miss the upcoming World Cup qualifiers against Georgia and Republic of Ireland.
"Bale - who missed Real Madrid's game against Espanyol on Sunday - arrived with the rest of the Wales squad on Sunday and attended Monday's annual FAW Awards dinner at the team headquarters in Cardiff.
"But, following dialogue with Real Madrid, Bale was sent for a scan and results confirmed that he wouldn't be fit to feature."
Bale has scored 26 goals in his 68 Wales appearances and played a huge role in their memorable run to the Euro 2016 semi-finals last summer.
Real boss Zinedine Zidane reported last weekend that Bale was suffering from "muscle fatigue", but added it was "nothing important".
"He could have played with us (against Espanyol) but I preferred to leave him out as he had a small complaint," Zidane said on Sunday.
"After so many games at the start of the season and four months of injury, he was suffering after the game in Dortmund."
Bale is still clearly feeling discomfort and his enforced absence could open the door for Liverpool teenager Ben Woodburn to make his first Wales start.
The 17-year-old made a dramatic debut last month, when he scored the winner within five minutes of coming on as a second-half substitute against Austria.
Wales are currently second in qualifying Group D and two wins are likely to guarantee them at least a place in next month's play-offs.
But they are only a point clear of third-placed Republic of Ireland, who they face in a potentially pivotal showdown on Monday night.