Eugenol Confers Cadmium Tolerance via Intensifying Endogenous Hydrogen Sulfide Signaling in Brassica rapa

Eugenol, a plant-derived small compound, shows great medicinal potential. However, whether and how eugenol regulates crop physiology remains elusive. Here we reported that eugenol induced Cd (cadmium) tolerance in the root of Brassica rapa. Roots were treated with eugenol and CdCl₂ simultaneously (eugenol + Cd) or pretreated with eugenol followed by CdCl₂ treatment (eugenol → Cd). Eugenol significantly attenuated Cd-induced growth inhibition, ROS accumulation, oxidative injury, and cell death, which were confirmed by in vivo histochemical analysis. Eugenol remarkably decreased free Cd²⁺ accumulation in root. Eugenol intensified GSH (glutathione) accumulation in roots upon CdCl₂ exposure, which explained the decrease in free Cd²⁺ and attenuation of oxidative injury. Eugenol stimulated endogenous H₂S (hydrogen sulfide) generation by upregulating the expression of BrLCD ( l-cysteine desulfhydrase) and BrDCD ( d-cysteine desulfhydrase) as well as their enzymatic activities in CdCl₂-treated root. Application of H₂S biosynthesis inhibitor or H₂S scavenger led to the decrease in endogenous H₂S level in Cd-treated root, which further compromised all the above effects of eugenol. These findings suggested that eugenol triggered H₂S → GSH signaling cassette in plants to combat Cd stress, which shed new light on eugenol-modulated plant physiology and the interaction between eugenol and H₂S.