The news, announced by Bolton on Wednesday, comes after Holdsworth’s company Sports Shield BWFC went into liquidation last week.
Sports Shield BWFC, which had owned 37.5 per cent of the shares in Wanderers’ parent company Burnden Leisure, and Ken Anderson’s Inner Circle Investments took over at the Macron Stadium in a joint venture in 2016.
A statement published on Bolton’s website on Wednesday said: “Burnden Leisure Limited can today confirm that with immediate effect Dean Holdsworth has resigned as a director from all group companies including Bolton Wanderers Football Club.
“No further comments in relation to this matter will be made.”
Holdsworth played for the club from 1997 until 2003.