Cryptic diversity in the estuarine copepod Acartia tonsa: reproductive isolation and lineage-specific divergence in transcriptomic response to salinity stress

Discovery of the near ubiquity of cryptic species among planktonic taxa marks a major advance in marine science over the last two decades. However, little is known about the potential drivers of divergence among species, and particularly how cryptic taxa may respond differentially to environmental stress as climate change and ocean acidification proceed. Here we report results from experimental studies characterizing reproduction and transcriptomic response to stress in Acartia tonsa, a dominant estuarine copepod that exhibits three sympatric (but morphologically cryptic) lineages that sort by salinity along the US Atlantic coast (lineage F - fresh, S - saline, and X - intermediate). Pair crosses between F and S individuals collected from Chesapeake Bay at a common ambient salinity (13ppt) revealed that while inter-lineage (FxS) crosses produced eggs, only eggs from intra-lineage (FxF or SxS) crosses successfully hatched. Analysis of whether 'hybrid' eggs were fertilized and failed to develop or did not fertilize, which could indicate pre vs post-zygotic isolation, is ongoing. Exposing replicate groups of 2^nd gen. lab-cultured F and S individuals to ambient and low salinity (7 ppt) stress produced very different patterns of gene expression measured with RNAseq. First, the dynamic range of expression was much greater in the S lineage overall (both salinities), with more differentially expressed genes (DEG’s) in comparisons of low vs control salinity (232 vs 77 DEGs at 0.01 FDR level for S, and F respectively). Gene ontology analysis of DEG function revealed a more targeted response in F to ion stress (up and down regulation of genes involved in ion transport, catalytic activity, and binding) while the S lineage showed a more varied and general response (up-regulation of metabolism, nucleic acid synthesis, and general stress response). Overall, these experiments confirm reproductive isolation among two cryptic A. tonsa ineages and identify key differences in thier physiological response to salinity stress. Future work will examine lineage-specific responses to other significant stressors such as hypoxia.