Long-Term Crop Rotation Improves Drought Resilience and Modulates Antioxidant Responses in Spring Wheat Leaves and Roots

Long-term crop rotation can improve soil function and crop performance, but it remains unclear whether rotation history can simultaneously alleviate drought-induced oxidative injury in spring wheat leaves and roots. To address this gap, we used a long-term field rotation experiment established in 2016 in the western foothills of the Greater Khingan Mountains. Four cropping systems were selected: spring wheat–potato rotation (R1), spring wheat–potato–rape rotation (R2), spring wheat–rape rotation (R3), and continuous spring wheat cropping (C1). In 2022, wheat occurred naturally in all rotation sequences, and all plots were planted with the same spring wheat cultivar (‘Longmai 36’) to enable comparison among different rotation histories. Drought stress was imposed from late jointing, and at anthesis, ROS-related levels, malondialdehyde (MDA), glutathione (GSH), and proline (Pro), as well as the activities of superoxide dismutase (SOD) and peroxidase (POD), were determined in the flag leaves and roots of spring wheat under normal-water (NC) and drought-stress (HC) conditions. Under drought stress, rotation significantly increased yield (R1 and R2 by 73.3% and 77.7% vs. C1, respectively, partly reflecting the low C1 baseline associated with continuous cropping) and reduced drought-induced accumulation of ROS-related levels and MDA, along with excessive antioxidant and osmotic responses. R1 better protected leaves, while R2 optimized root osmotic regulation. Two-way ANOVA revealed significant drought × rotation interactions for most leaf traits (p < 0.01) but not for root ROS-related levels, SOD, or POD, indicating organ-specific regulation. R1 and R2 show strong drought resilience potential, warranting multi-year, multi-site validation.