The US singer - famed for hits such as Crazy In Love and Single Ladies - will perform at the twin-site event in August, along with other bill-topping acts Kings Of Leon and double Brit-winner Emeli Sande.
Beyonce's performances will be her European festival appearances of the year.
She will be also playing two dates at the LG Arena in Birmingham in April.
V Festival - staged at Hylands Park, Chelmsford, and Weston Park in Staffordshire - is being staged over the weekend of August 17 and 18.
The Script, Jessie J, Rita Ora and X Factor winner James Arthur will also be playing V this summer.
The Vaccines, Beady Eye, Calvin Harris and Jessie Ware are also among the first batch of stars unveiled for the festival.
Festival director Simon Moran said: "With our line-up already boasting acts including Beyonce, Kings Of Leon and Emeli Sande, the 18th annual V Festival is once again set to be an unmissable weekend.
"We have a reputation for bringing fans the best world class acts and breakthrough artists, 2013 is set to be another sensational year."
In past headliners have included Coldplay, Radiohead, Oasis and Arctic Monkeys.
Further acts are to be announced over the coming weeks.
Tickets go on sale on Friday at 9am on www.VFestival.com.