Implication of quantifying nitrate utilization and CO₂ assimilation of Brassica napus plantlets in vitro under variable ammonium/nitrate ratios

Background: Plantlets grown in vitro with a mixed nitrogen source utilize sucrose and CO₂ as carbon sources for growth. However, it is very difficult to obtain the correct utilization proportions of nitrate, ammonium, sucrose and CO₂ for plantlets. Consequently, the biological effect of ammonium/nitrate utilization, the biological effect of sucrose/CO₂ utilization, and the ammonium/nitrate use efficiency for new C input derived from CO₂ assimilation/sucrose utilization are still unclear for plantlets.

Results: The bidirectional stable nitrogen isotope tracer technique quantified the proportions of assimilated nitrate and ammonium in Brassica napus plantlets grown at different ammonium/nitrate ratios. The utilization proportions of sucrose and CO₂ could be quantified by a two end-member isotope mixing model for Bn plantlets grown at different ammonium/nitrate ratios. Under the condition that each treatment contained 20 mM ammonium, the proportion of assimilated nitrate did not show a linear increase with increasing nitrate concentration for Bn plantlets. Moreover, the proportion of assimilated CO₂ did not show a linear relationship with the nitrate concentration for Bn plantlets. Increasing the nitrate concentration contributed to promoting the assimilation of ammonium and markedly enhanced the ammonium utilization coefficient for Bn plantlets. With increasing nitrate concentration, the amount of nitrogen in leaves derived from nitrate assimilation increased gradually, while the nitrate utilization coefficient underwent no distinct change for Bn plantlets.

Conclusions: Quantifying the utilization proportions of nitrate and ammonium can reveal the energy efficiency for N assimilation in plantlets grown in mixed N sources. Quantifying the utilization proportion of CO₂ contributes to evaluating the photosynthetic capacity of plantlets grown with variable ammonium/nitrate ratios. Quantifying the utilization proportions of nitrate, ammonium, sucrose and CO₂ can reveal the difference in the ammonium/nitrate use efficiency for new C input derived from CO₂ assimilation/sucrose utilization for plantlets grown at variable ammonium/nitrate ratios.