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The dynamics of nitrate (NO −  3 ), a major nitrogen (N) source for natural plants, has been studied mostly through experimental N addition, enzymatic assay, isotope labeling, and genetic expression. However, artificial N supply may not reasonably reflect the N strategies in natural plants because NO −  3  uptake and reduction may vary with external N availability. Due to abrupt application and short operation time, field N addition, and isotopic labeling hinder the elucidation of  in situ  NO −  3 -use mechanisms. The concentration and natural isotopes of tissue NO −  3  can offer insights into the plant NO −  3  sources and dynamics in a natural context. Furthermore, they facilitate the exploration of plant NO −  3  utilization and its interaction with N pollution and ecosystem N cycles without disturbing the N pools. The present study was conducted to review the application of the denitrifier method for concentration and isotope analyses of NO −  3  in plants. Moreover, this study highlights the utility and advantages of these parameters in interpreting NO −  3  sources and dynamics in natural plants. We summarize the major sources and reduction processes of NO −  3  in plants, and discuss the implications of NO −  3  concentration in plant tissues based on existing data. Particular emphasis was laid on the regulation of soil NO −  3  and plant ecophysiological functions in interspecific and intra-plant NO −  3  variations. We introduce N and O isotope systematics of NO −  3  in plants and discuss the principles and feasibilities of using isotopic enrichment and fractionation factors; the correlation between concentration and isotopes (N and O isotopes: δ 18 O and Δ 17 O); and isotope mass-balance calculations to constrain sources and reduction of NO −  3  in possible scenarios for natural plants are deliberated. Finally, we offer a preliminary framework of intraplant δ 18 O-NO −  3  variation, and summarize the uncertainties in using tissue NO −  3  parameters to interpret plant NO −  3  utilization.

Nitrate dynamics in natural plants: insights based on the concentration and natural isotope abundances of tissue nitrate