Stable isotope probing and oligotyping reveal the impact of organophosphorus pesticides on the carbon fixation related bacterioplankton lineage

Freshwater bacterioplankton communities play a pivotal role in global carbon fixation and energy exchange. However, establishing direct linkages between environmental stressors like organophosphorus pesticides (OPPs) and the ecological functions, such as carbon-fixing related microorganisms (CFMs), remains challenging. This study investigated the effects of four OPPs — two phosphates (dichlorvos, monocrotophos) and two phosphorothioates (omethoate, parathion) — on bacterioplankton communities using stable isotope probing, high-throughput sequencing and oligotyping analysis. Seven CFMs were identified. All OPPs significantly reduced total biomass (from 7.87 ×10⁴ to 2.30–4.11 ×10⁴ cells/mL) but stimulated CFMs proliferation. Notably, phosphorothioates induced a greater increase in CFMs abundance (36.84 %–57.18 %, up from 21.1 %) compared to phosphates (23.85 %–37.10 %; p < 0.05). Principal coordinate analysis (PCoA) revealed that phosphorothioates exerted stronger effects on microbial community and CFMs oligotypes structure compared to phosphates (p < 0.05). Variance partitioning analysis (VPA) identified pesticide type as the dominant driver of community structure. PICRUSt2 prediction demonstrated that OPPs suppressed oxidoreductase pathways linked to energy metabolism while activating transferase pathways associated with microbial stress resistance. Phosphorothioates depleted 64 pathways and enhanced 208 pathways, far exceeding phosphate impacts (2 depleted, 22 enhanced), indicating the phosphorothioates played a more important role on bacterioplankton communities than phosphate. Additionally, OPPs exposure reduced functional redundancy and destabilized community stability in bacterioplankton, potentially granting CFMs a long-term competitive advantage and elevating algal bloom risks. These findings provide insights into active CFMs in aquatic systems and their responses to diverse OPPs, offering new perspectives for managing organophosphorus pesticide contamination.