Increasing zinc (Zn) concentration in wheat grain is an important global challenge due to high incidence of Zn deficiency in human populations. In this study, a two-year field experiment was conducted to investigate the effects of foliar ZnSO₄ combined with various biostimulants (fulvic acid (FA), seaweed extract (SE), amino acids (AA), and microbial incubates (MI)) on Zn concentration and bioavailability in wheat grain under different soil nitrogen (N) levels (0, 120, and 240 kg N/ha). Grain Zn concentration and bioavailability were significantly enhanced by foliar Zn plus various biostimulants and soil N supply. Compared to foliar Zn alone, foliar Zn + FA resulted in 16% increase in grain Zn, mainly from insoluble Zn increases, while foliar Zn + AA caused 11% increase in grain Zn, mainly from soluble (at N0) and insoluble Zn increases (at N120). Foliar Zn + FA and Zn + AA generally resulted in higher Zn bioavailability than foliar Zn alone. Additionally, N concentration and Fe concentration and bioavailability in grain were enhanced with foliar Zn + AA and soil N application. Thus, foliar ZnSO₄ plus FA and AA under optimal soil N rate (120 kg N/ha) can be an effective and economically friendly approach for achieving agronomic biofortification.