Scalby Reserves quit Saturday League
The Otters, formed in 1912, have struggled to field two sides consistently so far this season after a host of players moved on in the summer.
The club's first team gained promotion from the second division last season, but a large number of their players decided to move on, leading to the club struggling to field two sides.
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Hide AdScalby FC secretary Phil Perry said: "Having a team in the first division means we can't continue to expect them to turn up with a bare 11.
"I wish to thank the team managers for all their efforts, they've worked tirelessly to try and keep two teams going, but in the end it wasn't possible."
Club treasurer Jez Clifford added: "It was always our intention to run two teams to give the youth setup a pathway into senior football, unfortunately it seemed that getting promoted to the first division was too big a challenge for some of last year's squad, so a number have either retired or moved on to easier pastures.
"Sadly the current squad size will not support two teams on a regular basis, despite huge efforts by the managers and club committee.
"We don't take this decision lightly and will try again next year."
Scalby's second string were due to play derby rivals FC Rosette on Saturday, but withdrew from the league beforehand.