Geobacter sulfurreducens is capable of reducing Pd(II) to Pd(0) using acetate as electron donor; however, the biochemical and genetic mechanisms involved in this process have not been described. In this work, we carried out transcriptome profiling analysis to identify the genes involved in Pd(II) reduction in this bacterium. Our results showed that 252 genes were upregulated while 141 were downregulated during Pd(II) reduction. Among the upregulated genes, 12 were related to energy metabolism and electron transport, 50 were classified as involved in protein synthesis, 42 were associated to regulatory functions and transcription, and 47 have no homologs with known function. RT-qPCR data confirmed upregulation of genes encoding PilA, the structural protein for electrically conductive pili, as well as c-type cytochromes GSU1062, GSU2513, GSU2808, GSU2934, GSU3107, OmcH, OmcM, PpcA, and PpcD under Pd(II)-reducing conditions. Delta pilA and Delta pilR mutant strains showed 20% and 40% decrease in the Pd(II)-reducing capacity, respectively, as compared to the wild type strain, indicating the central role of pili in this process. RT-qPCR data collected during Pd(II) reduction also confirmed downregulation of omcB, omcC, omcZ, and omcS genes, which have been shown to be involved in the reduction of Fe(III) and electrodes. The present study contributes to elucidate the mechanisms involved in Pd(II) reduction by G. sulfurreducens. Graphical