The present experiment was conducted to investigate the effects of dietary supplementation with different levels of betaine on nutrient utilization and digestive enzyme activity in indigenous yellow-feather broilers during starter phase. A total of 288 male, one-d-old Huaixiang chickens with an average initial body weight (BW) of 44.52 ± 1.07 g were randomly allotted to 4 treatments. Each dietary treatment consisted of 6 replicates, with 12 birds per replicate. The 4 dietary treatments were corn-soybean meal-based diets and supplemented with 0, 500, 1000 and 2000 mg/kg betaine, respectively. The trial period lasted 28 d. It was found that dietary betaine supplementation improved the calcium (Ca) utilization (cubic, P=0.0187). Meanwhile, dietary supplementation with different levels of betaine increased the trypsin activity (quadratic, P=0.0445) and lipase activity (cubic, P=0.0344) of the jejunum. Taken together, this study revealed that betaine could promote the digestive function in indigenous yellow-feather broilers during starter phase.