*Article* **The E3 Ubiquitin Ligase Gene** *Sl1* **Is Critical for Cadmium Tolerance in** *Solanum lycopersicum* **L.**

**Chen-Xu Liu 1, Ting Yang 1, Hui Zhou 1, Golam Jalal Ahammed 2,\*, Zhen-Yu Qi <sup>3</sup> and Jie Zhou 1,4,5,\***


**Abstract:** Heavy metal cadmium (Cd) at high concentrations severely disturbs plant growth and development. The E3 ubiquitin ligase involved in protein degradation is critical for plant tolerance to abiotic stress, but the role of E3 ubiquitin ligases in Cd tolerance is largely unknown in tomato. Here, we characterized an E3 ubiquitin ligase gene *Sl1*, which was highly expressed in roots under Cd stress in our previous study. The subcellular localization of Sl1 revealed that it was located in plasma membranes. In vitro ubiquitination assays confirmed that Sl1 had E3 ubiquitin ligase activity. Knockout of the *Sl1* gene by CRISPR/Cas9 genome editing technology reduced while its overexpression increased Cd tolerance as reflected by the changes in the actual quantum efficiency of PSII photochemistry (ΦPSII) and hydrogen peroxide (H2O2) accumulation. Cd-induced increased activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were compromised in *sl1* mutants but were enhanced in *Sl1* overexpressing lines. Furthermore, the content of Cd in both shoots and roots increased in *sl1* mutants while reduced in *Sl1* overexpressing plants. Gene expression assays revealed that Sl1 regulated the transcript levels of heavy metal transport-related genes to inhibit Cd accumulation. These findings demonstrate that Sl1 plays a critical role in regulating Cd tolerance by relieving oxidative stress and resisting heavy metal transportation in tomato. The study provides a new understanding of the mechanism of plant tolerance to heavy metal stress.

**Keywords:** antioxidant enzymes; heavy metal stress; ubiquitination; protein degradation; tomato
