Host selection and nutrient status jointly drive algal and bacterial interactions in epiphytic biofilms of submerged macrophytes: Structural and functional insights

Epiphytic biofilms play a crucial role in aquatic biogeochemical cycles but are simultaneously influenced by host selection and eutrophication. However, the compositional structure and interaction mechanisms of these factors on algal and bacterial communities remain poorly understood. In this study, we employed Confocal Laser Scanning Microscopy (CLSM), Scanning Electron Microscopy (SEM), and high-throughput sequencing to investigate the physicochemical properties, algal and bacterial diversity, and community structure of epiphytic biofilms on two submerged macrophytes - Vallisneria natans (VN) and Hydrilla verticillata (HV) - across three urban shallow lakes with varying trophic levels in the Yangtze River Basin. Our results revealed distinct algal and bacterial communities influenced by both host plants and lake nutrient conditions, with unique core species identified in VN, HV, and the surrounding water. Host-environment effects index (HEEI = 1.79) indicated that bacterial communities were predominantly shaped by host selection, exhibiting lower diversity in HV (1.66 ± 0.92) and VN (2.31 ± 1.12) biofilms compared to surrounding waters (3.80 ± 0.47). In contrast, algal communities were primarily regulated by environmental factors (HEEI = 0.43), with higher diversity in less eutrophic lakes. Algal-bacterial co-occurrence network analysis revealed greater network complexity in VN biofilms than that in HV, with predominantly synergistic interactions facilitating carbon and nitrogen cycling. Eutrophication increased biofilm thickness, nutrient content, and extracellular polymeric substance (EPS) production but reduced microbial diversity and altered community interaction patterns. This study advances our understanding of epiphytic biofilms and offers insights into optimizing host-microbe interactions for improving lake restoration strategies.