Previous animal studies found beneficial effects of choline and betaine on maternal glucose metabolism during pregnancy, but few human studies explored the association between choline or betaine intake and incident gestational diabetes mellitus (GDM). We aimed to explore the correlation of dietary choline or betaine intake with GDM risk among Chinese pregnant women. A total of 168 pregnant women with GDM cases and 375 healthy controls were enrolled at the Seventh People’s Hospital in Shanghai during their GDM screening at 24–28 gestational weeks. A validated semi-quantitative FFQ was used to estimate choline and betaine consumption through face-to-face interviews. An unconditional logistic regression model was adopted to examine OR and 95 % CI. Compared with the controls, those women with GDM incidence were likely to have higher pre-pregnancy BMI, be older, have more parities and have higher plasma TAG and lower plasma HDL-cholesterol. No significant correlation was observed between the consumption of choline or betaine and incident GDM (adjusted OR (95 % CI), 0·77 (0·41, 1·43) for choline; 0·80 (0·42, 1·52) for betaine). However, there was a significant interaction between betaine intake and parity on the risk of GDM (Pfor interaction = 0·01). Among those women with no parity history, there was a significantly inverse correlation between betaine intake and GDM risk (adjusted OR (95 % CI), 0·25 (0·06, 0·81)). These findings indicated that higher dietary betaine intake during pregnancy might be considered a protective factor for GDM among Chinese women with no parity history.