Comprehensive Analysis of Metabolome and Transcriptome Reveals Physiological Processes Related to Larval Development of Barnacles (<i>Megabalanus volcano</i>).

Abstract

<h4>Background</h4>Barnacles are important marine fouling organisms, and their complex life cycle involves key metamorphic nodes from nauplius to cyprid larvae and then to sessile adults. However, the molecular mechanisms underlying their larval development remain poorly understood. Metabolomics and transcriptomics are powerful tools for exploring biological development pathways and regulatory networks.<h4>Methods</h4>We employed non-targeted metabolomics and transcriptomics to analyze three key developmental stages of embryonic stage, nauplius stage, and cyprid stage. Differential metabolites were screened using fold change (FC), <i>p</i>-value, and variable importance in projection (VIP) values, while DEGs were identified with adjusted <i>p</i>-value and |log₂(fold change)| criteria. WGCNA was used to construct gene co-expression networks, and qRT-PCR validated RNA-seq results.<h4>Results</h4>A total of 3683 metabolites were identified, with the bile secretion pathway serving as a core regulatory pathway throughout early development. Transcriptomic analysis identified 7234 DEGs, which were clustered into four modules corresponding to different developmental stages. Key pathways such as chitin metabolism, and linoleic acid metabolism were significantly enriched, and qRT-PCR confirmed the reliability of RNA-seq data.<h4>Conclusions</h4>This study reveals the metabolic and molecular regulatory mechanisms underlying the early development of <i>M. volcano</i>, highlighting stage-specific metabolic characteristics and core gene modules. The findings provide a theoretical basis for understanding barnacle developmental adaptation strategies and offer potential targets for the development of novel antifouling agents.