# Coexpression among eastern oyster host and microbiome genes suggests coordinated regulation of calcifying fluid chemistry

## TL;DR

Oysters regulate their shell-forming fluid chemistry, and their microbes may help by influencing pH and carbonate levels, suggesting a host-microbe partnership in calcification.

## Contribution

Reveals coordinated gene expression between oysters and their microbiome in regulating calcifying fluid chemistry.

## Key findings

- Oyster calcifying fluid pH remains stable despite tidal fluctuations, with increased ion transport and acid-base regulation gene expression.
- Microbial nitrogen and sulfur cycling genes correlate with higher dissolved inorganic carbon in calcifying fluid.
- Coexpression networks link oyster immune and neural pathways to microbial redox processes, suggesting host-microbe interaction.

## Abstract

Oysters and many marine animals build shells by controlling the chemistry of extracellular fluids where minerals form, yet whether microbes in these fluids influence calcification remains unclear. We show that oysters maintain favorable conditions for mineral formation by regulating the carbonate chemistry of the shell-forming fluid, and that resident microbes respond to these changes by expressing nitrogen- and sulfur-cycling genes capable of altering pH, alkalinity, and carbonate availability. Many of these microbial transcripts were tightly correlated with oyster immune and signaling genes, suggesting that host and microbiome processes may be linked within the calcifying environment. These findings point to a host–microbiome interaction in the regulation of calcifying-fluid chemistry that directly links microbial activity to the carbonate chemistry underlying biomineralization.

Marine animals that build shells, such as oysters, carefully regulate the chemistry of their internal calcifying fluids, but the molecular mechanisms behind this control, as well as whether microbes play a role in calcification, are poorly understood. To better understand oysters’ molecular mechanisms and the role of their calcifying-fluid microbes, we conducted experiments that simulated a tidal cycle, measured calcifying fluid pH and total dissolved inorganic carbon, and characterized host and microbial gene expression via transcriptomics. These experiments showed that calcifying fluid pH remained relatively stable throughout tidal pH fluctuations, with corresponding increases in oyster transcripts for ion transport and acid–base regulation. These data provide direct evidence that tidal fluctuations drive rapid changes in oyster calcifying fluid chemistry. Most surprisingly, increases in microbial transcripts related to nitrogen and sulfur cycling correlated to higher calcifying fluid DIC, and coexpression network analysis revealed patterns of gene expression that linked oyster immune and neural pathways to microbial redox processes, providing molecular evidence of potential host modulation of microbial metabolism. Together, these results reveal that oysters actively regulate their calcifying fluid pH over short timescales, and the endemic microbiome metabolic responses can yield metabolites that influence calcifying fluid pH, alkalinity, and ultimately calcification. These data offer a perspective on oyster physiological capacity and, most importantly, the potential role of microbes in oyster calcification. In light of ongoing changes in ocean pH and temperature, oysters provide a model for studying animal–microbial responses to environmental acidification and how their interactions may shape biomineralization.