Marketa Vondrousova's Wimbledon title defence came to an early end as she fell to a surprise first-round defeat by Spain's Jessica Bouzas Maneiro.
The Czech became the first unseeded player to win the women's singles trophy when she triumphed against Ons Jabeur in last year's final.
But Vondrousova's game was riddled with unforced errors and double faults as the 25-year-old opened proceedings with a lacklustre performance on Centre Court.
World number 83 Bouzas Maneiro, 21, took advantage on Tuesday to win 6-4 6-2 for her first Grand Slam victory.
It is the first time since 1994, when Steffi Graf lost to Lori McNeil, that the women's defending champion has lost in the first round at Wimbledon.
"I am really happy, I think this is one of the most important moments in my life, in my career," said Bouzas Maneiro.
"This is the most beautiful tournament I have played in my life so thank you to everyone who came to watch the match today."
Vondrousova's early exit means there will be a different women's singles champion for a seventh consecutive year.
Serena Williams was the last player to successfully defend the Venus Rosewater Dish, winning back-to-back titles in 2015 and 2016.
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