Mr Guthrie's fortune is stated as £313m, an increase in value of £3m on last year's figure.
He and his family own Scarborough-based property group Broadland Properties, based in Scalby Road.
The 32nd annual edition of the Rich List says the "jewel" in the Guthrie empire is Hever Castle in Kent, Anne Boleyn's childhood home.
It reveals that the profits of Broadland Properties fell to £6.7m on £49.8m revenue in 2018.
Mr Guthrie, 83, and his family are listed at 413 in the list of the country's 1,000 richest people.
Mr Guthrie holds the OBE for his charity work. He was a major benefactor to Scarborough Rugby Union Club - its clubhouse bears the name JM Guthrie but that is in honour of his father, as he did not want the recognition - after Sport England turned down the club's application for funding.
He is a chartered surveyor by profession and founded Broadland Properties in 1950.
Broadland have interests throughout Britain and Poland, ranging from agricultural and sporting estates to commercial premises and developments. It purchased Hever Castle, in Kent, and its collection from the Astor family in 1983.
As well as the rugby club, Mr Guthrie has supported many organisations, including Saint Catherine's Hospice, Scarborough Museums Trust, Stephen Joseph Theatre, The Rainbow Centre, Ryedale Festival, Scarborough Symphony Orchestra and Wykeham Parochial Church Concert.
The 2020 Sunday Times Rich List is based on identifiable wealth, including land, property, other assets such as art and racehorses, or significant shares in publicly quoted companies. It excludes bank accounts, to which the paper has no access.
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