Emma Raducanu pulled out of her mixed doubles match with Andy Murray on Saturday, bringing the 37-year-old's Wimbledon career to an abrupt end.
Raducanu withdrew citing stiffness in her right wrist, with the 21-year-old due to play in the fourth round of the singles on Sunday afternoon.
It means Murray's defeat alongside brother Jamie in the men's doubles on Thursday night was his last ever match at the Championships.
Murray's career was celebrated in a Centre Court ceremony on Thursday (PA)
The two-time champion cannot re-pair with another player, according to Wimbledon rules.
Murray is said to be disappointed. His mother, Judy Murray, replied "Yes, astonishing" to a post on Raducanu's decision on Twitter/X.
Raducanu was pictured training in an England top on Saturday afternoon before her decision to withdraw from the mixed doubles was announced.
Murray's Wimbledon career was celebrated in a special ceremony on Centre Court following his defeat in the men's doubles alongside brother Jamie, but Murray believed he had another match to come as he waved farewell.
Raducanu, the world number 135, was in sensational form as she defeated Maria Sakkari in the singles on Friday to reach the fourth round but the 21-year-old said in a statement: "Unfortunately I woke up with some stiffness in my right wrist this morning.
"Therefore I have decided to make the very tough decision to withdraw from the mixed doubles tonight.
"I'm disappointed as I was really looking forward to playing with Andy but got to take care."
Raducanu celebrates beating world number nine Maria Sakkari (PA)
Murray asked Raducanu to be his partner for the mixed doubles on his final Wimbledon appearance after he was forced to withdraw from the singles.
The champion in 2013 and 2016 decided he had not recovered from an operation on a spinal cyst in time to compete in the singles competition.
The pairing of Murray and Raducanu, Britain's only grand slam singles champions this century, promised to be a treat for Wimbledon fans.
Raducanu accepted Murray's offer and said she was looking forward to playing with the former champion at Wimbledon.
"For me, it's a once-in-a-lifetime opportunity," she said following her victory in the second round. "I think some things are bigger than just tennis.
The names of Murray and Raducanu are removed from the order of play board (Getty)
"At the end of my life, at the end of my career when I'm like 70 years old, I know I'm going to have that memory of playing Wimbledon with Andy Murray on a home slam."
Murray and Raducanu's match was scheduled fourth on Court One on Sunday, which potentially would not have started until late in the evening.
Raducanu is scheduled to face qualifier Lulu Sun, the world 123, for a place in the quarter-finals on Sunday in what has been her best grand slam performance since winning the US Open three years ago.
Raducanu, who took a wildcard into the main draw, wishes to prioritise the singles and said she was "disappointed" to to make the "very tough decision" to withdraw.
Murray's Wimbledon career was celebrated in a special event on Centre Court on Thursday following his defeat in the men's doubles alongside brother Jamie.
Sue Barker led the ceremony in front of Murray's family and peers. The 37-year-old was watched by wife Kim, his oldest daughters, eight-year-old Sophia and six-year-old Edie, mother Judy and father Willie.
Murray wipes away tears before reminiscing with Sue Barker (PA)
Murray's fiercest rivals Roger Federer, Rafael Nadal and Novak Djokovic also took part in a video message, while Venus Williams highlighted Murray's support and advocacy of women's tennis.
Murray could not hold back the tears as he turned to his team and said he wanted to "play forever" while speaking to Barker on court.
"It is hard because I would love to keep playing but I can't," he said. "Physically it's just too tough now. All of the injuries have added up and they haven't been insignificant.
"I want to play forever. I love this sport. It's given me so much, taught me loads of lessons over the years that I can use for the rest of my life. I don't want to stop, so it is hard."
Murray has entered the Paris Olympics, where he has a place in the singles and will play in the doubles with Dan Evans before ending his career.
