Abstract
An epilithic algal matrix (EAM) exhibits rapid expansion, recovery capacity, and high adaptability, leading to widespread distribution in degraded coral reef habitats. However, limited research on the dynamic processes of succession hinders a comprehensive understanding of EAM formation. To examine the influence of succession processes and environmental factors on the composition of EAM microbial communities, a three-factor (time × depth × attached substrate type) crossover experiment was conducted in the Luhuitou Reef Area, Sanya, China. Microbial community compositions were analyzed through 16S rRNA gene amplicon sequencing. The community was predominantly composed of proteobacteria (61.10-92.75%), cyanobacteria (2.47-23.54%), bacteroidetes (0.86-8.49%), and firmicutes (0.14-7.76%). Successional processes were found to significantly shape the EAM-associated microbial communities in the Luhuitou Reef Area. Proteobacteria played a crucial role in biofilm formation during this process, while cyanobacteria contributed to the structural complexity of microhabitats within the EAM. A chaotic aggregation stage of approximately one month was observed before transitioning into an expansion stage, eventually stabilizing into a low-diversity community. Although the relatively smooth substrate supported high biodiversity, microorganisms displayed no preference for the three different substrates. While no significant differences in community composition were observed at small-scale depths, cyanobacteria and bacteroidetes showed positive correlations with light and temperature, respectively. The EAM-associated microbial community exhibited higher complexity in the shallower regions under increased light intensity and temperature. Given the characteristics of the microbial community succession process, continuous monitoring of changes in microbial community structure and key taxa (such as proteobacteria and cyanobacteria) during EAM formation is recommended.