Heavy metal pollution and soil salinization harm human health and the environment. Phytoremediation is a widely accepted soil decontamination method, with woody plants being particularly effective due to their large biomass and extensive root systems. In this study, we identified and cloned PsnMLP328 from Populus simonii x P. nigra and demonstrated its role in mitigating salt and cadmium stress. PsnMLP328 expression was up-regulated under both stress conditions, and its overexpression in tobacco enhanced resistance to these stresses, albeit through distinct mechanisms. Transgenic plants exhibited increased Cd2+ uptake and a higher biomass, alleviating Cd2+-induced growth inhibition. Additionally, PsnMLP328 boosted proline content, chlorophyll levels, and antioxidative enzyme activities (POD, SOD) under Cd2+ stress, likely by protecting cells from oxidative damage. Expression analysis revealed that PsnMLP328 down-regulated the cadmium transporter Nramp2 while up-regulating YSL2 (another cadmium transporter) and potassium channels (AKT1 and AKT2/3), suggesting its role in modulating K+ and Cd2+ homeostasis. These findings indicate that PsnMLP328 enhances tobacco resistance to salt and cadmium stress, particularly the latter. This study is the first to elucidate the function of poplar MLP family genes under salt and cadmium stress, advancing our understanding of MLP gene roles in heavy metal stress and offering new insights for remediating salinized and heavy metal-contaminated soils.