Enzymatic Activity Responses to Transport and Low-Temperature Storage: Implications for Plant Nitrogen Metabolism Studies

Understanding how transport and storage conditions affect enzymatic activity is essential for accurate biomonitoring of nitrogen metabolism in plants. This study evaluated the effects of transport conditions and low-temperature storage on the enzymatic activities of nitrate reductase (NR), glutamine synthetase (GS), and phosphomonoesterase (PME) for Chloris gayana, Fraxinus uhdei, and Trifolium repens. Enzymatic activities were measured for leaf samples immediately after collection, after 18 h at room temperature, or after 18 h on ice. Additionally, samples were stored at −16 °C or −45 °C for up to 28 days. NR activity decreased to near-zero levels under all storage conditions, indicating that this enzyme is unsuitable for delayed analysis. In contrast, GS and PME activities showed species-dependent responses to storage, with increased activity observed for T. repens and C. gayana, potentially reflecting tissue degradation processes. F. uhdei exhibited greater stability in enzyme activities, suggesting a higher resilience to storage. These findings highlight the importance of minimizing storage time to preserve enzymatic integrity, particularly for NR, while providing insight into the potential for delayed analysis of GS and PME in specific species. This work offers practical recommendations for future biomonitoring efforts in nitrogen deposition studies.