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Children's white matter development is driven by experience, yet it remains poorly understood how it is shaped by attending formal education. A small number of studies compared children before and after the start of formal schooling to understand this, yet they do not allow to separate maturational effects from schooling‐related effects. A clever way to (quasi‐)experimentally address this issue is the longitudinal school cut‐off design, which compares children who are similar in age but differ in schooling (because they are born right before or after the cut‐off date for school entry). We used for the first time such a longitudinal school cut‐off design to experimentally investigate the effect of schooling on children's white matter networks. We compared "young" first graders (schooling group, n = 34; Mage = 68 months; 20 girls) and "old" preschoolers (non‐schooling group, n = 33; Mage = 66 months; 18 girls) that were similar in age but differed in the amount of formal instruction they received. Our study revealed that changes in fractional anisotropy and mean diffusivity in five a priori selected white matter tracts during the transition from preschool to primary school were predominantly driven by age‐related maturation. We did not find specific schooling effects on white matter, despite their strong presence for early reading and early arithmetic skills. The present study is the first to disentangle the effects of age‐related maturation and schooling on white matter within a longitudinal cohort of 5‐year‐old preschoolers. Research Highlights: White matter tracts that have been associated with reading and arithmetic may be susceptible to experience‐dependent neuroplasticity when children learn to read and calculate.This longitudinal study used the school cut‐off design to isolate schooling‐induced from coinciding maturational influences on children's white matter development.White matter changes during the transition from preschool to primary school are predominantly driven by age‐related maturation and not by schooling effects.Strong effects of schooling on behavior were shown for early reading and early arithmetic, but not for verbal ability and spatial ability. [ABSTRACT FROM AUTHOR]