Nanochitin whisker (NC) with a cationic nature could enhance plant photosynthesis, grain yield, and quality of wheat, but have not been systematically studied. This study was designed to investigate the stimulatory effects of NC on dry matter (DM) and nitrogen (N) accumulation and translocation, and on the metabolism of carbon (C) and N in later growth stages of winter wheat to reveal the enhancement mechanism of grain yield and crude protein concentration. Different parts of NC-treated plants from pot grown experiments were collected at the pre- and post-anthesis stages. The accumulation, translocation, and contributions of DM and N from pre-anthesis vegetation organs to grains, as well as key metabolic enzyme activities, including sucrose phosphate synthase (SPS) and phosphoenolpyruvate carboxylase (PEPC), were examined. The results showed that, at an application rate of 6 mg·kg−1 of NC in the soil, the accumulation of DM and N were significantly enhanced by 16.2% and 38.8% in pre-anthesis, and by 15.4% and 30.0% in post-anthesis, respectively. Translocation of N and DM in the post-anthesis periods were enhanced by 38.4% and 50.9%, respectively. NC could also stimulate enzyme activities, and increased 39.8% and 57.1% in flag leaves, and by 36.0% and 58.8% in spikes, respectively, at anthesis. SPS and PEPC increased by 28.2% and 45.1% in flag leaves, and by 42.2% and 56.5% in spikes, respectively, at 15 days after anthesis. The results indicated that the NC promoted N metabolism more than C metabolism, and resulted in the enhancement of grain yield by 27.56% and of crude protein concentration in grain by 13.26%, respectively.

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