Mass spectrometry imaging enables visualization of the localization of glutamate decarboxylase activity in germinating legume seeds

Visualizing the distribution of enzymes is vital for understanding physiological phenomena. Enzyme histochemistry is a technique used to investigate the localization of enzyme activity. However, the target is restricted to enzymes with easy-to-design artificial substrates that can develop color through reactions. Mass spectrometry imaging (MSI)-based enzyme histochemistry has been developed as a novel method to visualize enzyme localization. It can be applied to enzyme histochemistry as it detects products from the supplied substrate using enzymes present on the tissue sections. However, enzyme histochemistry using MSI has not been applied to plant tissue samples yet. Glutamate decarboxylase (GAD, EC: 4.1.1.15) is an enzyme that catalyzes the decarboxylation reaction of l-glutamic acid to produce γ-aminobutyric acid (GABA). GABA biosynthesis is important both in the field of food chemistry and plant physiology. This study focused on GAD during the legume germination process and successfully visualized GAD activity in legume seeds using MSI for the first time. Furthermore, the localization of GAD activity in the embryonic axis of germinated soybean seeds and alfalfa seeds could be visualized. •Mass spectrometry imaging was used to visualize enzymatic activity on plant samples for the first time.•Enzyme localization was visualized using MSI in soybean and alfalfa seeds.•Highest enzyme activity detected at 60 °C at a pH of 5.8.•GAD activity was highest in germinated seeds compared to non-germinated seeds.•Enzymes had a constant reaction rate when sufficient substrate was available.