The goal of this work was to define resistant and susceptible variables of young broccoli (Brassica oleracea L. convar. botrytis (L.) Alef. var. cymosa Duch.) plants treated with cold and hot water. Additionally, we wanted to single out variables that could potentially be used as biomarkers of cold/hot-water stress in broccoli. Hot water changed more variables (72%) of young broccoli than cold water (24%) treatment. Hot water increased the concentration of vitamin C for 33%, hydrogen peroxide for 10%, malondialdehyde for 28%, and proline for 147%. Extracts of broccoli stressed with hot water were significantly more efficient in the inhibition of α-glucosidase (65.85 ± 4.85% compared to 52.00 ± 5.16% of control plants), while those of cold-water-stressed broccoli were more efficient in the inhibition of α-amylase (19.85 ± 2.70% compared to 13.26 ± 2.36% of control plants). Total glucosinolates and soluble sugars were affected by hot and cold water in an opposite way, which is why they could be used as biomarkers of hot/cold-water stress in broccoli. The possibility of using temperature stress to grow broccoli enriched with compounds of interest to human health should be further investigated.