Infant temperament is associated with excess weight gain or childhood obesity risk in samples of healthy individuals, although the evidence has been inconsistent. To our knowledge, no prior research has examined this topic among children exposed to gestational diabetes mellitus (GDM) in utero. To prospectively evaluate infant temperament in association with overweight and obesity status at ages 2 to 5 years among children born to mothers who experienced GDM. This prospective cohort study took place at Kaiser Permanente Northern California medical centers. We studied singleton infants delivered at 35 weeks’ gestational age or later to mothers who had been diagnosed with GDM. Data were collected from 2009 to 2016, and data analysis occurred from June 2017 to October 2018. The primary exposures in the child’s first year were soothability, distress to limitations, and activity aspects of temperament, as assessed by a validated questionnaire. Modifiable covariates in the child’s first year included breastfeeding intensity and duration monthly ratio scores, along with the timing of the introduction of sugary beverages and complementary foods. The primary outcome was child overweight and obesity status, assessed at ages 2 to 5 years. Multinomial logistic regression models estimated adjusted odds ratios and 95% CIs for infants whose temperaments were measured at 6 to 9 weeks of age and categorized as elevated (≥75th percentile) or not elevated in the 3 domains. We controlled for nonmodifiable and modifiable covariates across models. A total of 382 mother-infant pairs participted, including 130 infants (34.0%) who were non-Hispanic white, 126 infants (33.0%) who were Hispanic, 96 infants (25.1%) who were Asian, 26 infants (6.8%) who were non-Hispanic black, and 4 infants (1.1%) who were of other races/ethnicities. In descriptive analyses, elevated infant soothability and activity temperaments were associated with the early introduction of 100% fruit juice and/or sugar-sweetened beverages (at ages <6 months) and shorter breastfeeding duration (from 0 to <3 months), while elevated distress to limitations was associated with early introduction of complementary foods (at ages <4 months). Elevated soothability consistently was associated with a higher odds of later childhood obesity, with adjusted odds ratios across models ranging from 2.22 (95% CI, 1.04-4.73) to 2.54 (95% CI, 1.28-5.03). Greater breastfeeding intensity and duration (12-month combined) score was associated with lower odds of obesity, independent of infant temperament and other covariates. Among this high-risk population of infants, elevated soothability was associated with early childhood obesity risk, perhaps in part because caregivers use sugary drinks to assuage infants. Soothability temperament may be a novel screening target for early obesity prevention interventions involving responsive feeding and emotion regulation.