Sedentary time is associated with increased risk of type 2 diabetes, but the association between objectively measured sedentary time and incident gestational diabetes mellitus (GDM) has not been tested. The purpose of this paper is to test associations between objectively measured sedentary time and self-reported television time during pregnancy with incident GDM and plasma glucose levels among women at high risk for GDM.At 20 weeks' gestation, pregnant women (n = 188) in the North East of England with a risk factor for GDM wore an activPAL accelerometer and reported their usual television time. Participants underwent a standard oral glucose tolerance test at 24-28 weeks' gestation. Regression analyses were used to test for associations of total and prolonged sedentary time, breaks in sedentary time, and television time with GDM and fasting and 2-h glucose levels. Interaction terms were applied to examine whether the association between each indicator of sedentary time and glucose levels differed by GDM status.Total sedentary time (hours/day) was not associated with incident GDM (OR 1.00 (95%CI 1.00, 1.01)). The association between total sedentary time and glucose levels depended on GDM status: sedentary time was associated with fasting (β = 0.16 (95%CI 0.01, 0.31)) and 2-h (β = 0.15 (95%CI 0.01, 0.30)) glucose levels for those without GDM, while breaks in sedentary time were associated with lower fasting (β = - 0.55 (95%CI - 0.92, - 0.17)) and 2-h (β = - 0.40 (95%CI - 0.77, - 0.03)) glucose levels for those with GDM. Prolonged sedentary time was associated with higher fasting glucose levels regardless of GDM status (β 0.15 (0.01, 0.30)). Television time was associated with development of GDM (OR 3.03 (95%CI 1.21, 7.96)) but not with plasma glucose levels.This is the first study to test associations between posture-based measures of sedentary time during pregnancy and GDM and glucose levels. The findings presented here suggest the possible importance of minimizing or breaking up sedentary time for the management of glucose levels during pregnancy, at least among women at high risk of GDM. Further research is needed to understand the different roles of total sedentary time and television time in the development of GDM.