Species-Specific Effects of Humic Substances and Mycorrhiza on Antioxidant Defense and Metal Stress Tolerance in Cannabis sativa, Sorghum sudanense × bicolor, and Miscanthus × giganteus Under Field Conditions

Abstract

Abiotic stresses, including heavy metal contamination, can severely impair plant growth and antioxidative defense. However, their adverse effects may be mitigated through sustainable strategies such as biostimulant application. This study investigated the effects of humic substances (HSs), alone or combined with mycorrhizal inoculation (M), on oxidative stress and antioxidative responses in Cannabis sativa, Sorghum sudanense & times; bicolor, and Miscanthus & times; giganteus grown under field conditions on metal-contaminated agricultural soil exceeding regulatory thresholds for Zn, Pb, and Cd. Plant growth, lipid peroxidation, stress-related metabolites (proline, sugars), antioxidative enzyme activities (catalase, CAT; ascorbate peroxidase, APX; guaiacol peroxidase, GOPX; glutathione reductase, GR, and superoxide dismutase, SOD), and leaf metal concentrations were analyzed. Biostimulants increased proline and sugars in Sorghum (by up to 55% and 80%, respectively), accompanied by reduced oxidative stress indicators and improved biomass (by 26%). In Cannabis, higher Cd and Pb concentrations following biostimulant treatments were associated with increased SOD, APX, and GR activities (by 33-267%), without affecting growth. In Miscanthus, increased lipid peroxidation (by 37-60%) occurred alongside enhanced GR and APX activities. These results indicate strong species-specific responses and absence of consistent synergistic effects of HSs and M, highlighting distinct physiological strategies of stress adaptation and antioxidative defense on metal-contaminated soils. Future research should address physiological and molecular mechanisms underlying these responses.