Entertainment Weekly reports that the actor will play "a mystery man" with a connection to Chuck's mother (Linda Hamilton).
The episode will mark the former James Bond star's first US television role since an appearance in Charlie's Angels in 1979.
"We're huge fans of Mr. Dalton, not only for being a bad-ass Bond but for his comic work in Hot Fuzz and The Rocketeer," revealed producer Josh Schwartz. "In truth, we've wanted to have him on the show since season 1. When we came up with this part, we thought he'd be perfect."
However, Schwartz dismissed speculation that Dalton's character would be revealed to be the real father of Chuck (Zachary Levi).
"Consider that theory squashed," he said. "We'll only say that [he and Chuck's mother have] known each other for a long time."
The actor will make his first appearance in the seventh episode of the new season, titled 'Chuck Versus the First Fight'.
Chuck returns to NBC on September 20.