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Microbial communities play a fundamental role in biogeochemical cycling within salt and brackish marsh ecosystems, yet fungal‐prokaryotic interactions in these environments remain poorly understood. This study employed metabarcoding of the 16S and 28S rRNA genes to investigate prokaryotic and fungal communities across four locations in sediments and surface waters of the North Inlet salt marsh and Winyah Bay brackish marsh (South Carolina, USA) over four time points from 2020 to 2021. Co‐occurrence network analyses were used to identify potential microbial interactions and their ecological implications. Distinct fungal and prokaryotic communities were observed between the two marsh types. From the 16S prokaryotic primer set, Proteobacteria, Bacteroidota, and Cyanobacteriota dominated both marshes. Early diverging fungi and Actinomycetota (bacteria) were prevalent in the brackish marsh, whereas salt marsh communities were primarily composed of Dikarya fungi (Ascomycota and Basidiomycota) and Desulfobacteria. Network analyses revealed contrasting interactions between surface water and sediment. In brackish marsh sediments, fungi and prokaryotes exhibited nearly exclusively negative connections, suggesting strong resource competition. In contrast, Dikarya fungi in brackish marsh surface water displayed numerous positive connections with bacteria, suggesting potential cross‐feeding interactions. In the salt marsh, fungi and prokaryotes exhibited potential cooperative and competitive/antagonistic interactions. Ascomycota were positively connected with Desulfobacteria, suggesting a role in complex organic matter degradation. Conversely, negative connections between Chytridiomycota (early diverging fungi) and Cyanobacteriota (bacteria) implied parasitic interactions. These findings highlight the dynamic nature of fungal‐prokaryotic interactions in coastal ecosystems. By analyzing potential microbial relationships in salt and brackish marshes, this study provides new insights into the ecological roles of fungi in estuarine environments, particularly their contributions to nutrient cycling and organic matter decomposition. Understanding these interactions is crucial for generating hypotheses and predicting microbial responses to environmental changes—such as shifts in salinity and nutrient availability—and their potential impacts on marsh ecosystem functioning.
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Beyond dikarya: 28S metabarcoding uncovers cryptic fungal lineages across a tidal estuary
Fungi are key drivers of biogeochemical processes, yet marine fungi remain understudied and under-characterized due to primer biases and database gaps. In this study, we conducted a metabarcoding survey targeting the small and large subunit rRNA genes and the internal transcribed spacer region of fungi (18S, 28S, and ITS2) in the sediment and surface water of salt and brackish marshes in the North Inlet—Winyah Bay estuarine system (Georgetown, South Carolina, USA). The universal 18S/16S primer set (515F-Y and 926R) identified few fungal taxa. The ITS2 primer set (ITS3mix and ITS4) revealed high diversity among Dikarya but failed to capture the full extent of early diverging fungi (EDF). In contrast, the 28S primer set (LR0R and LF402) excelled at identifying EDF lineages, including Chytridiomycota, Mucoromycota, Zoopagomycota, and Blastocladiomycota, many of which dominated the brackish marsh sampling site but were less prevalent in the salt marsh sampling sites. Over half of the fungal OTUs identified by the 28S primer set were from EDF lineages. Copy-normalized 28S qPCR showed that EDF were more abundant in brackish sediments than in the salt marsh. Several putative denitrifying fungi, primarily species from Trichoderma and Purpureocillium, were also detected, suggesting overlooked functional guilds that may contribute to estuarine nitrogen cycling. A FUNGuild analysis found that most lineages were saprotrophic. Overall, our findings show that EDF are key contributors to community differences across salinity gradients and may play more important functional roles in coastal biogeochemistry than is currently understood. The 28S primer set is ideal for marine fungal metabarcoding because it provides comprehensive taxonomic coverage and enables phylogenetic analysis.
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- Award ID(s):
- 2303089
- PAR ID:
- 10644125
- Publisher / Repository:
- Springer Nature
- Date Published:
- Journal Name:
- Environmental Microbiome
- Volume:
- 20
- Issue:
- 1
- ISSN:
- 2524-6372
- Page Range / eLocation ID:
- 129
- Subject(s) / Keyword(s):
- Marine fungi Metabarcoding Salt marsh Brackish marsh Early diverging fungi Dikarya
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript
- Journal Article:
- https://doi.org/10.1186/s40793-025-00786-3
