Mercury (Hg) contamination of aquatic systems has been recognized as a global, serious problem affecting both wildlife and humans. High levels of Hg, in particular methylmercury (MeHg), were detected in surface sediments of Tagus Estuary. MeHg is neurotoxic and its concentration in aquatic systems is dependent upon the relative efficiency of reduction, methylation, and demethylation processes, which are mediated predominantly by the microbial community, in particular mercury-resistant (HgR) bacteria. Plants in contaminated ecosystems are known to take up Hg via plant roots. Therefore, the aims of this study were to (1) isolate and characterize HgR bacteria from a salt marsh of Tagus Estuary (Rosário) and (2) determine HgR bacteria levels in the rhizosphere and, consequently, their influence in metal cycling. To accomplish this objective, sediments samples were collected during the spring season in an area colonized by Sacocornia fruticosa and Spartina maritima and compared with sediments without plants. From these samples, 13 aerobic HgR bacteria were isolated and characterized morphologically, biochemically, and genetically, and susceptibility to Hg compounds, Hg2+, and MeHg was assessed by determination of minimal inhibitory concentration (MIC). Genetically, the mer operon was searched by polymerase chain reaction (PCR) and 16S rRNA sequencing was used for bacterial identification. Results showed that the isolates were capable of growing in the presence of high Hg concentration with MIC values for HgCl2 and MeHgCl in the ranges of 1.7–4.2 μg/ml and 0.1–0.9 μg/ml, respectively. The isolates from sediments colonized with Sacocornia fruticosa displayed higher resistance levels compared to ones colonized with Spartina maritima. Bacteria isolates showed different capacity of Hg accumulation but all displayed Hg volatilization capabilities (20–50%). Mer operon was found in two isolates, which genetically confirmed their capability to convert Hg compounds by reducing them to Hg0. Thus, these results are the first evidence of the relevance of interaction between bacteria and plants in Hg cycling in Tagus Estuary.