The aim of the research was to compare the impact of short-term deprivation of selected macronutrients (Ca, K, Mg and P) on the photosynthetic characteristics, transpiration and pigment content in maize. The strongest inhibition of photosynthesis was caused by a deprivation of Mg, which was visible as a decrease in the photosynthetic and transpiration rates, stomatal conductance, photosystem II (PSII) performance, chlorophyll and flavonol content with a simultaneously increased content of anthocyanins. In the K-deprived plants, a decrease in the photosynthetic rate was observed. However, the transpiration rate and stomatal conductance did not differ significantly compared with the control. In the K-deprived plants, a decrease in chlorophyll and an increase in the anthocyanin content were also observed. We showed that Ca starvation resulted in a decrease in the photosynthetic and transpiration rates, stomatal conductance and PSII performance, while the pigment content was not significantly different compared with the control. In the case of P-deprived plants, we observed a decrease in the photosynthetic and transpiration rates. Interestingly, the inhibition of stomatal conductance was the strongest in the P-deprived plants compared with all of the investigated elements. However, the performance of PSII was not significantly affected by P starvation compared with the control. Our results present for the first time a comprehensive analysis of the effect of short-term macronutrient deprivation on photosynthesis and transpiration in maize plants.