Williamson went down early in Wednesday’s 1-0 defeat to Manchester United and is the third Arsenal player to suffer the injury, alongside Beth Mead and Vivianne Miedema.
It means the 26-year-old will miss the World Cup in Australia this summer.
An Arsenal club statement read: “We can confirm that Leah Williamson suffered a ruptured anterior cruciate ligament in our Barclays Women’s Super League match against Manchester United on Wednesday night.
“Leah was substituted in the first half of the game at Leigh Sports Village and underwent further assessment on Thursday to determine the extent of the injury.
“Leah will now begin a period of rehabilitation and is set for an extended spell on the sidelines. She will undergo surgery in due course.
“Everyone at Arsenal will be supporting Leah closely throughout the journey ahead and we would ask that her privacy is respected at this time.”
Williamson is now the 11th player in the Women’s Super League to suffer an ACL injury.
Arsenal teammate Mead sent a message of support saying: “We are with you every step of the way @leahcwilliamson even if I have to be quiet some days in rehab.”
The World Cup kicks off on July 20, with European champions England having Haiti, Denmark and China in their group.
Meanwhile, Arsenal will now travel to Germany for their Champions League semi-final first leg tie at Wolfsburg without captain Kim Little and vice-captain Williamson.
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