The former Bond girl, 62, cited irreconcilable differences in the legal documents, which were filed last week in L.A.'s County Superior Court, according to TMZ.
Jane has requested joint legal and physical custody of their 17-year-old twin sons, Johnny and Kristopher. She is not requesting spousal support.
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The British actress and her actor-director husband James Keach, 65, announced they were ending their 20-year marriage in a joint statement in April.
"The couple confirm that they are separated and have been for several months," a rep for Jane and James told People magazine. "At this time they are negotiating the terms of their divorce."
The statement added, "They will continue their relationship as devoted parents to their children, as business associates and partners, and in their joint dedication to preserving and furthering the charitable endeavours that they've worked on throughout their marriage."
Days after their separation was announced, the stars put on a united front at the Open Hearts Foundation fundraiser as they posed with charity's beneficiaries.
This will be the Dr. Quinn, Medicine Woman star's fourth divorce. Her first marriage in 1971 to Michael Attenborough, son of director Lord Attenborough, ended after two years. She then wed artist Geoffrey Planer in 1977, but they split a year later.
Her third marriage to David Flynn came to an end after 11 years. The pair had two children, Katherine and Sean, together.
Jane met her current husband James on the set of Dr. Quinn. The couple wed in 1993.