This study aimed to explore the difference between two Chinese local broilers, one slow- and one fast-growing, in their response to a stress challenge. We conducted the study on slow- (Weining chicken) and fast-growing (Jinlinghua chicken) breeds, with 50 chickens from each breed either feed restricted to 70% for 30 days as a stress or given ad libitum to evaluate the effects on behavior, corticosterone, and microbial programming. Standing behavior was more frequent while exploration was less common in fast-growing breeds compared to slow-growing breeds. Food seeking and ingestion, exploration, and drinking increased, while resting decreased in the feed restricted treatments. There was no difference in corticosterone concentration between slow- and fast-growing chickens, but the level was affected by feeding treatments, and the interaction of breed and feed restriction. At the genus-level, the relative abundance of Bacteroides and Lactobacillus was higher, while Cloacibacillus and Megasphaera was lower in the slow-growing breed compared to the fast-growing breed. Feed restricted birds had a higher abundance of Mucispirillum, but lower abundance of Cloacibacillus, Clostridium XlVa and Clostridium IV. In conclusion, feed restriction to 70% for 30 days as a chronic stress stimulation caused more activity, elevated the stress response, and altered gut microbiota composition, but some changes were only evident in slow- or fast-growing chickens.