Seed germination is a complex process involving several stages, starting with the imbibition of water and ending with the emergence of the radicle. In the current study, we address the observation of an unexpected pH shift during the imbibition of maize grains. We used direct pH measurements of soak water, the pH indicator methyl red, and anatomical analysis to shed light on the acidification associated with maize (Zea mays L.) germination, a largely overlooked phenomenon. Our work shows that acidification during imbibition of maize grains is a two-step process: (i) early, rapid acidification (pH values up to 4.4), in which protons stored in the (dead) pericarp/testa are mobilised and rapidly diffuse into the surrounding medium, and (ii) late, delayed acidification (pH values just below 6), starting hours after contact of grains with water, representing an active transport process caused by living cells of the seed. We discuss the physiological mechanisms and ecological relevance of environmental acidification during maize grain germination.