Judicious application of nitrogen (N) fertilizers in crop production is critical for reducing the nitrate pollution of groundwater and greenhouse gas emissions. It is, thus, important to improve the nitrogen use efficiency under the reduced application of nitrogen. A genotypic variation in N-uptake and N-use efficiency particularly under low N-input conditions exists across crops that can be deciphered and exploited for environmentally sustainable farming without any significant penalty of yield and quality. The present research conducted under the nutrient solution culture aimed to explore the inherent variability in the growth response of ten genetically diverse wheat varieties to low fertilizer N-application (N-, 10 μM N) in comparison to N sufficient control (N+, 8.5 mM N) viz., a viz., the activity of various key N-assimilating enzymes and to delineate the indirect effect of low N on uptake and partitioning of other major macronutrients viz., P, K, S, which may indirectly regulate the N-use efficiency. A notable increase in sulfur, potassium, and phosphorus content was observed under nitrogen-deficient conditions. Varieties such as Carnamah and HD 2824 exhibit a significant increase in shoot phosphorus content, emphasizing their potential to optimize phosphorus acquisition and utilization efficiency under nutrient-limited conditions. The findings highlight the complex interplay between nutrient availability and plant responses, showcasing varietal-specific adaptations to nitrogen limitations.