Abstract It is unknown how hypohydration influences fine motor performance training and motor learning. Here, 30 participants (aged 19–46 years) were randomly assigned to a hypohydration (HYPO) or control (CON) group (both n = 15). Moderate hypohydration (~ 2.4% loss in body mass) was produced in HYPO via active dehydration before a 46 min fluid restricted rest period was undertaken. The conclusion of rest coincided with when CON attended the facilities. Both groups undertook a discrete sequence production task consisting of 6 training blocks, and returned ~ 300 min later to complete a delayed retention and transfer test while euhydrated. Bilateral pre-frontal cortex (PFC) haemodynamics were assessed using functional near-infrared spectroscopy throughout training and delayed learning assessments. Response time improved across training ( P textless 0.01) and was similar between the groups (both P = 0.22). Analysis of training PFC haemodynamics revealed a significant group by block interaction for oxygenated (O 2 Hb; P textless 0.01), but not deoxygenated haemoglobin ( P = 0.77). In training block 1, bilateral O 2 Hb was higher in HYPO ( P = 0.02), while bilateral O 2 Hb increased in CON between blocks 2–3 and 5–6 (both P ≤ 0.03). During the delayed retention and transfer test, no group differences or interactions were found in response time, response error, or PFC haemodynamics (all P ≥ 0.27). Moderate hypohydration does increase PFC activation during motor skill learning, however, this appears to be transient and of little consequence to training or delayed retention or transfer performance.