Reading is a cognitive skill that requires our brain to go through a myriad of changes during learning. While many studies have described how reading acquisition shapes childrens' brain function, less is known about the impact of reading on brain structure. Here we examined short-term causal effects of reading training on preschoolers' behavior and white matter structure. Forty-eight English-speaking preschoolers (4y10m to 6y2m) participated in a randomized controlled trial where they were randomly assigned to two training programs: the Letter training program was focused on key skills for reading (e.g., decoding and letter knowledge), while the Language training program strengthened oral language comprehension skills without exposure to text. Longitudinal behavioral data showed that only the Letter Training group increased letter knowledge and decoding skills after the two-week training. Diffusion MRI measures (FA and MD) of eighteen white matter pathways (including the left arcuate and the left inferior longitudinal fasciculus) did not reveal any statistically significant changes for either group despite high degrees of scan-rescan reliability across sessions. These findings suggest that a two-week reading training program can cause changes in preschoolers' letter knowledge and decoding abilities, without being accompanied by measurable changes in the diffusion properties of the major white matter pathways of the reading network. We conclude highlighting possible constraints (i.e., age, training onset and duration, cognitive profile) to reading-related white matter plasticity.