Use of gaseous 13NH3 administered to intact leaves of Nicotiana tabacum to study changes in nitrogen utilization during defence induction

Nitrogen‐13 (t_1/2 9.97 m), a radioactive isotope of nitrogen, offers unique opportunities to explore plant nitrogen utilization over short time periods. Here we describe a method for administering ¹³N as gaseous ¹³NH₃ to intact leaves of Nicotiana tabacum L. (cv Samsun), and measuring the labelled amino acids using radio high‐performance liquid chromatography (HPLC) on tissue extract. We used this method to study the effects of defence induction on plant nitrogen utilization by applying treatments of methyl jasmonate (MeJA), a potent defence elicitor. MeJA caused a significant increase relative to controls in key ¹³N]amino acids, including serine, glycine and alanine by 4 h post‐treatment, yet had no effect on ¹³NH₃ incorporation, a process that is primarily under the control of the glutamine synthatase/glutamate synthase pathway (GS/GOGAT) in cellular photorespiration. We suggest that the reconfiguration of nitrogen metabolism may reflect induction of non‐photorespiratory sources of nitrogen to better serve the plant's defences.