Five-time Wimbledon champion Venus Williams became the eighth of the top 10 seeds to be knocked out after a third-round defeat by Kiki Bertens.
Ninth seed Williams, 38, was beaten 6-2 6-7 (4-6) 8-6 by the 26-year-old Dutchwoman on Court One.
Williams’ fellow American Madison Keys lost nine consecutive games in her defeat to qualifier Evgeniya Rodina.
The 10th seed lost 7-5 5-7 6-4 in two hours 10 minutes against the Russian, who is ranked 120th in the world.
However, younger sister Serena to reach the last 16.
Venus Williams made a sluggish start to the contest and her body language in the first set belied her frustration.
At 5-1 Williams, who won Wimbledon in 2000, 2001, 2005, 2007 and 2008, saved a set point on her serve to avoid being broken for a third time.
However, 20th seed Bertens remained unflustered and clinically closed out the set with an ace.
World number nine Williams eventually found some rhythm in the second set as the pair twice traded breaks – Beterns missing a chance to serve out for the match at 5-4 up.
In the tie-break that followed, Williams won two mini breaks to have a 6-2 advantage only to fluff a chance to close out the set before she eventually levelled the match.
The pair exchanged early breaks in the third set before Bertens squandered two break points in the eighth game as Williams held.
Williams saved two match break points on deuce at 7-6 down, but the pressure told and she struck a return low into the net to hand Bertens the victory in two hours and 40 minutes.