Adrian Prout faced Cheltenham magistrates yesterday accused of killing his 55-year-old wife Kate Prout, even though her body has never been found.
Prout, dressed in a pale green jumper and a white shirt, spoke only to confirm his name and address.
Magistrates refused him bail and he was remanded in custody by magistrates and will reappear at Bristol Crown Court next Tuesday.
The 46-year-old still lives at Redhill Farm in Redmarley, near Newent, the home he shared with his wife until she vanished on November 5, 2007.
A huge police search involving nearly 100 officers was launched, during which forensic experts spent days searching 220 acres of fields and woodland around the farm.
Divers and specially-trained dogs [Eddie and Keela, specialist cadaver scent & blood trained dogs which took part in the hunt for Madeleine McCann ] were used in a bid to find traces of the missing retired teacher but failed to discover anything.
Mr Prout was first arrested on suspicion of murder in November 2007. He was re-arrested in February 2008 and his police bail was then cancelled in January this year.
But police arrested him again at Redhill Farm yesterday morning and charged him his wife's murder.
Ann Reddrop, head of the Crown Prosecution Service complex casework unit, said: "We have been working closely with the police over the past months to investigate all aspects of Mrs Prout's disappearance before reaching a final decision on charging.
"We have also sought advice from leading counsel. Accordingly, we have this week authorised the police to charge Mr Prout."
Senior investigating officer acting detective superintendent Neil Kelly added: "This has been a complex, thorough and necessarily lengthy investigation to bring us to this point.
"I would like to take this opportunity to thank all those affected, especially Mrs Prout's friends and family, for their understanding, tolerance and patience."
The Crown Prosecution Service said yesterday that they had sufficient evidence for a murder trial. Mr Prout was remanded by Cheltenham JPs until March 17.
Mrs Prout spent 20 years as a primary school teacher before taking early retirement. At the time of her disappearance she was involved in a campaign to persuade the Government to increase milk payments to dairy farmers.
Eddie and Keela - Top sniffer dogs
Used across the world for his accuracy, seven-year-old hound Eddie helped police put Trevor Hamilton behind bars in 2006 after the victim recovery dog found 63-year- old Attracta Harron's blood on the 23-year-old murderer's burned-out Hyundai.
Eddie, who works for South Yorkshire police, also located Attracta's body in a shallow grave in April 2003.
Last year the dog and his handlers returned to Ulster for a third time to help find missing Arlene Arkinson.
The Tyrone teenager went missing after leaving a disco in Bundoran, Co Donegal, on August 13, 1994.
Both Eddie and Keela have also been used in various disappearance and murder cases in the US and the Republic of Ireland.
Eddie and Keela versus Team McCann
Eddie and Keela, the two sniffer British dogs used to find evidences of Madeleine's death, in Portugal were used by the police in the searches of Kate Prout. Although investigations at her Redhill Farm home finished a fortnight ago it is understood specialist sniffer dogs used in the search for missing four-year-old Madeleine McCann in Portugal have been used in the investigation and found evidence of Mrs Prout's death.