Peer Review #3 of "Optimized nitrogen fertilizer application strategies under supplementary irrigation improved winter wheat (Triticum aestivum L.) yield and grain protein yield (v0.1)"

Background.Exploring suitable split nitrogen management is essential for winter wheat production in the Huang-Huai-Hai Plain of China (HPC) under water-saving irrigation conditions, which can increase grain and protein yields by improving nitrogen translocation, metabolic enzyme activity and grain nitrogen accumulation.Methods.Therefore, a 2-year field experiment was conducted to investigate these effects in HPC.Nitrogen fertilizer was applied at a constant total rate (240 kg/ha), split between the sowing and at winter wheat jointing growth stage in varying ratios, N1 (0% basal and 100% dressing fertilizer), N2 (30% basal and 70% dressing fertilizer), N3 (50% basal and 50% dressing fertilizer), N4 (70% basal and 30% dressing fertilizer), and N5 (100% basal and 0% dressing fertilizer).Results.We found that the N3 treatment significantly increased nitrogen accumulation post-anthesis and nitrogen translocation to grains.In addition, this treatment significantly increased flag leaf free amino acid levels, and nitrate reductase and glutamine synthetase activities, as well as the accumulation rate, active accumulation period, and accumulation of 1000-grain nitrogen.These factors all contributed to high grain nitrogen accumulation.Finally, grain yield increase due to N3 ranging from 5.3% to 15.4% and protein yield from 13.7% to 31.6%.The grain and protein yields were significantly and positively correlated with nitrogen transport parameters, nitrogen metabolic enzyme activity levels, grain nitrogen filling parameters.Conclusions.Therefore, the use of split nitrogen fertilizer application at a ratio of 50%:50% basaltopdressing is recommended for supporting high grain protein levels and strong nitrogen translocation, in pursuit of high-quality grain yield.