Oxidative stress reshapes diatom-microbiome interactions by shifting benefits from mutualistic to opportunistic bacteria

Abstract The roles of reactive oxygen species (ROS) as signaling molecules and inhibitors of phytoplankton growth are well documented. While phytoplankton physiological mechanisms for ROS detoxification are well characterized, the role of heterotrophic bacterial partners in ROS alleviation and outcomes for these bacteria remain poorly understood. Here, we examined how extracellular hydrogen peroxide (H 2 O 2 ) shapes nutrient exchange between the diatom Phaeodactylum tricornutum and two phycosphere bacteria. From an initial screen of 20 bacteria, we identified a “helper” ( Muricauda sp.) that enabled P. tricornutum to survive acute H 2 O 2 stress and a “non-helper” ( Algoriphagus sp.) that did not. Using nanoscale secondary ion mass spectrometry (nanoSIMS), we tracked diatom-derived carbon and nitrogen ( 13 C and 15 N) transfer to each bacterial partner under ROS stress. Oxidative stress disrupted diatom metabolism and altered nutrient transfer: diatom-derived carbon and nitrogen incorporation was significantly reduced in the helper but increased in the non-helper under H 2 O 2 stress. Growth assays revealed that the helper preferentially utilized exudates from healthy, intact hosts, whereas the non-helper did not grow on exudates but thrived on lysates from damaged or lysed cells. Together, these findings indicate the helper was better adapted to accessing resources from living hosts, while the non-helper relied on nutrients released through ROS-induced host damage. Our results highlight oxidative stress as a key driver of algal-bacterial interactions and suggest that bacterial resource-acquisition strategy underlies host protection: bacteria utilizing healthy-host exudates are more likely to protect hosts from oxidative stress, while those benefiting from host damage are not, despite retaining ROS detoxification capacity.