The mass ‘conkerthon’ saw 502 visitors to the Royal Horticultural Society's garden at Wisley, Surrey, taking part in 251 matches.
Players of all ages travelled from right across the south of England to help set a new Guinness World Record.
They included 64-year-old reigning World Conker Champion Ray Kellock, who has put his successful technique down to a maths formula.
The previous record of 197 matches played by 394 people was set by Bristol’s Redland High School for Girls in 2006.
The RHS organised last month’s conkers caper to highlight the plight of the horse chestnut.
RHS chief horticultural advisor Guy Barter said: "This is a fantastic achievement. Well done to all of the 'conkerers' who helped to set this new record.
"It clearly shows the great affection with which the common horse chestnut is held."