N uptake, assimilation and isotopic fractioning control δ 15N dynamics in plant DNA: A heavy labelling experiment on Brassica napus L.

In last decades, a large body of evidence clarified nitrogen isotope composition (delta N-15) patterns in plant leaves, roots and metabolites, showing isotopic fractionation along N uptake and assimilation pathways, in relation to N source and use efficiency, also suggesting N-15 depletion in plant DNA. Here we present a manipulative experiment on Brassica napus var. oleracea, where we monitored delta N-15 of purified, lyophilized DNA and source leaf and root materials, over a 60-days growth period starting at d 60 after germination, in plants initially supplied with a heavy labelled (delta N-15(Air-N2) = 2100 mUr) ammonium nitrate solution covering nutrient requirements for the whole observation period (470 mg N per plant) and controlling for the labelled N species (H-4, O-3 and both). Dynamics of Isotopic Ratio Mass Spectrometry (IRMS) data for the three treatments showed that: (1) leaf and root delta N-15 dynamics strictly depend on the labelled chemical species, with H4O3 and H4O3 plants initially showing higher, lower and intermediate values, respectively, then converging due to the progressive NH4+ depletion from the nutrient solution; (2) in H4O3, where delta N-15 was not affected by the labelled chemical species, we did not observe isotopic fractionation associated to inorganic N uptake; (3) delta N-15 values in roots compared to leaves did not fully support patterns predicted by differences in assimilation rates of NH4+ and NO3-; (4) DNA is depleted in N-15 compared to the total N pools of roots and leaves, likely due to enzymatic discrimination during purine biosynthesis. In conclusion, while our experimental setup did not allow to assess the fractionation coefficient (epsilon) associated to DNA bases biosynthesis, this is the first study specifically reporting on dynamics of specific plant molecular pools such as nucleic acids over a long observation period with a heavy labelling technique.

Automated summary

This study conducted a manipulative experiment on Brassica napus var. oleracea to monitor the dynamics of δN-15 in purified DNA, source leaf, and root materials over a 60-day growth period. The results showed that the δN-15 dynamics of leaves and roots depend on the labeled chemical species, with some unexpected findings regarding isotopic fractionation and N-15 depletion in plant DNA. While the study did not assess the fractionation coefficient (epsilon) associated with DNA bases biosynthesis, it provides valuable insights into the dynamics of specific plant molecular pools over a long observation period using heavy labelling techniques.