X-ray absorption spectroscopy is a well-established method for probing local structural and electronic atomic environments in a variety of systems. We used X-ray absorption near-edge structure (XANES) spectroscopy for monitoring in real-time conditions selenium reduction in situ in live cultures of Shewanella oneidensis MR-1 under high hydrostatic pressure. High-quality XANES data show that Shewanella oneidensis MR-1 reduces selenite Se(IV) to red elemental selenium Se(0) up to 150 MPa without any intermediate redox state. MR-1 reduces all selenite provided (5-10 mm) between 0.1 and 60 MPa. Above 60 MPa the selenite reduction yield decreases linearly with pressure and the activity is calculated to stop at 155 +/- 5 MPa. The analysis of cultures recovered after in situ measurements showed that the decrease in activity is linked to a decrease in viability. This study emphasizes the promising potential of XANES spectroscopy for real-time probing in situ microbial redox transformations of a broad range of metal and metalloid elements in live samples, including under high hydrostatic pressure.