Succession pattern and phylotype analysis of microphytobenthic communities in a simulated oil spill seagrass mesocosm experiment

Abstract

Microphytobenthic communities play a significant role in nutrient modulation, sediment stabilization, and primary production in seagrass beds, which provide various ecosystem services. We hypothesized that microphytobenthic communities in sediments of chronically oil-exposed seagrass beds will exhibit increased resiliency to stressors associated with oil exposure as opposed to seagrass beds never exposed to oil spills. We prepared 14-liter seawater mesocosms, each containing a submersed macrophyte Ruppia maritima collected from the Chandeleur Islands, Louisiana, and Estero Bay, Florida. Mesocosms were initially exposed to 50% water-accommodated oil fractions (WAF) and subsequently diluted by 50% with daily artificial seawater exchanges over 8 days to simulate tidal dilution. High-throughput amplicon sequencing based on 23S rRNA gene targeting cyanobacteria and chloroplasts of eukaryotic microphytobenthos was conducted to assess the impact of oiling on microphytobenthic communities with additional assessment via microscopy. High-throughput sequencing in combination with traditional microscopic analysis provided a robust examination in which both methods roughly complemented each other. Distinct succession patterns were detected in benthic algal communities of chronically oil-exposed (Louisiana) versus unexposed (Florida) seagrass bed sediments. The impact of oiling in microphytobenthos across all samples showed that benthic diatoms dominated all algal communities with sample percentages ranging from 42 to 97%, followed by cyanobacteria (2 to 50%). It is noteworthy that drastic changes in microphytobenthic community structure in terms of the larger taxonomic level were not observed, rather change occurred at the phylotype level. These results were also confirmed by microscopy. Similarity percentages (SIMPER) analysis identified seven phylotypes (Cyanobacteria, Bacillariophyceae, and Mediophyceae) in the Louisiana samples and one phylotype (Bacillariophyceae) in the Florida samples that increased in relative sequence abundance after oil exposure. The detailed phylotype analysis identifying sentinel microphytobenthic indicators provides a base for future research on benthic microalgae response to ecosystem disturbance.

Keywords: 23S rRNA; Amplicon sequencing; Benthic microalgae; Deepwater horizon; Disturbance ecology; Macondo oil spill; Ruppia maritima.