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HighlightsThis work utilizes “multi-stage pulse labeling”  15 N applications, primarily during reproductive growth stages, as a phenotyping strategy to identify maize hybrids with superior N use efficiency (NUE) under low N conditions.Research using labeled isotopic N ( 15 N) can precisely quantify fertilizer nitrogen (N) uptake and organ-specific N allocation in field crops such as maize ( Zea mays  L.). The overall research objective was to study plant N uptake patterns potentially correlated with N use efficiency (NUE) in field-grown maize hybrids using a “multi-stage pulse labeling”  15 N phenotyping strategy with an emphasis on the reproductive period. Five hybrids varying in NUE were compared under zero N fertilizer application (0N) plus a moderate rate of 112 kg N ha −1  (112N) in 2013 (2 locations) and 2014 growing seasons. The equivalent of 3.2 (2013) to 2.1 (2014) kg of  15 N ha −1 , as labeled Ca( 15 NO 3 ) 2 , was injected into soil on both sides of consecutive plants at multiple stages between V14 and R5. Aboveground plant biomass was primarily collected in short-term intervals (4–6 days after each  15 N application) in both years, and following a single long-term interval (at R6 after  15 N injection at R1) in 2014. Averaged across hybrids and site-years, the moderate N rate (112N) increased absolute  15 N uptake at all stages; however, plants in the 0N treatment allocated proportionally more  15 N to reproductive organs. Before flowering, short-term recovery of  15 N ( 15 Nrec) totaled ~0.30 or 0.40 kg kg −1  of the  15 N applied, and ~50% of that accumulated  15 Nu was found in leaves and 40% in stems. After flowering, plant  15 Nrec totaled ~0.30 kg kg −1  of  15 N applied, and an average 30% of accumulated  15 Nu was present in leaves, 17% in stems, and the remainder—usually the majority—in ears. At the R5 stage, despite a declining overall rate of  15 N uptake per GDD thermal unit, plant  15 Nrec represented ~0.25 kg kg −1  of  15 N applied, of which ~65% was allocated to kernels. Overall long-term  15 Nrec during grain filling was ~0.45 and 0.70 kg kg −1  of total  15 N applied at R1 with 0 and 112N, respectively, and most (~77%)  15 N uptake was found in kernels. The “multi-stage pulse labeling” technique proved to be a robust phenotyping strategy to differentiate reproductive-stage N uptake/allocation patterns to plant organs and maize efficiencies with newly available fertilizer N.

Applicability of a “Multi-Stage Pulse Labeling” 15N Approach to Phenotype N Dynamics in Maize Plant Components during the Growing Season