<p>Bioerosion by boring sponges, particularly those from the Clionaidae family, plays a crucial role in the breakdown of calcium carbonate (CaCO<sub>3</sub>) substrates in coastal ecosystems such as coral and oyster reefs. The molecular mechanisms driving this process, particularly the expression of carbonic anhydrase, a key enzyme hypothesized to aid in CaCO<sub>3</sub> dissolution, remain poorly understood. Here we have developed a new genetic method for detecting carbonic anhydrase expression in <i>Cliona celata</i> to evaluate bioerosion at the molecular level. A pair of PCR primers targeting carbonic anhydrase genes was designed and tested on <i>C. celata</i> specimens under predator simulation and control conditions. Another novel primer pair was designed to target heat shock protein 70, which does not change in expression during bioerosion, to function as a control. RNA was extracted and analyzed using PCR and qPCR to assess gene expression levels. The results confirmed successful amplification of the carbonic anhydrase gene, demonstrating the efficacy of the primers. While no significant difference in gene expression was observed between treatments, the study establishes a foundational tool for further molecular exploration of sponge bioerosion. The developed primers have the potential for broader application across the Clionaidae family, offering a valuable method for advancing our understanding of bioerosion mechanisms in sponges. Future research should focus on validating the primer across different sponge species and exploring its utility in studies correlating gene expression with phenotypic bioerosion activity.</p>

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A genetic approach to bioerosion: developing a novel primer pair targeting carbonic anhydrase genes in a Clionaid boring sponge

  • Indiana Olson,
  • Melissa DeBiasse,
  • Amber D. Stubler,
  • Morgan Kelly

摘要

Bioerosion by boring sponges, particularly those from the Clionaidae family, plays a crucial role in the breakdown of calcium carbonate (CaCO3) substrates in coastal ecosystems such as coral and oyster reefs. The molecular mechanisms driving this process, particularly the expression of carbonic anhydrase, a key enzyme hypothesized to aid in CaCO3 dissolution, remain poorly understood. Here we have developed a new genetic method for detecting carbonic anhydrase expression in Cliona celata to evaluate bioerosion at the molecular level. A pair of PCR primers targeting carbonic anhydrase genes was designed and tested on C. celata specimens under predator simulation and control conditions. Another novel primer pair was designed to target heat shock protein 70, which does not change in expression during bioerosion, to function as a control. RNA was extracted and analyzed using PCR and qPCR to assess gene expression levels. The results confirmed successful amplification of the carbonic anhydrase gene, demonstrating the efficacy of the primers. While no significant difference in gene expression was observed between treatments, the study establishes a foundational tool for further molecular exploration of sponge bioerosion. The developed primers have the potential for broader application across the Clionaidae family, offering a valuable method for advancing our understanding of bioerosion mechanisms in sponges. Future research should focus on validating the primer across different sponge species and exploring its utility in studies correlating gene expression with phenotypic bioerosion activity.