Summary of the range of exposure duration, acclimation time, time per trial, recovery period, and sample size for copper and blue rockfish used to test behavioral and physiological responses to elevated <i>p</i>CO₂.

<p>Note: Individual fish were used successively in the different trials to enable tracking of performance measures. Data from fish that did not behave normally in a particular trial were excluded (e.g., refusal to swim in the U_crit test). In addition, 2 of 12 blue rockfish individuals that were sequenced had low quality reads and were subsequently excluded from the differential gene expression analysis.</p

Mean carbonate chemistry conditions (± standard error) in the experimental system.

<p>Shown are mean values of dissolved organic carbon (DIC), total alkalinity, pH (total scale), <i>p</i>CO₂, and temperature.</p

Enzyme activity ratios for lactate dehydrogenase and citrate synthase (LDH:CS) in white muscle tissue of (A) copper and (B) blue rockfish following chronic exposure to extreme <i>p</i>CO₂ (~2800 μatms) or control (~550 μatms) treatments.

<p>Copper rockfish exhibited a significant increase in aerobic enzyme activity relative to anaerobic activity (i.e., lower LDH:CS ratio) from the control to extreme high <i>p</i>CO₂ treatments (two-way ANOVA; Species×Treatment: <i>F</i>_{<i>1</i>,<i>24</i>} = 4.46, <i>P</i> = 0.045), whereas blue rockfish did not differ in LDH:CS activity among treatments.</p

Gene expression profiles (A) and Gene Ontology (GO) functional categories (B) for copper and blue rockfish muscle tissue as a function of <i>p</i>CO₂ treatment.

<p>(A) Heatmaps display significant differential gene expression (DE) for copper (<i>n</i> = 147) and blue (<i>n</i> = 358) rockfish (FDR<0.001) among <i>p</i>CO₂ treatments; green = up-regulation, red = down-regulation. Each column represents an individual fish (<i>n</i> = 15 copper rockfish and <i>n</i> = 10 blue rockfish). Genes are ordered by similarity in gene expression profile and differ in both order and identity between the two species (only 14 DE genes were in common between the two species). Hierarchical (Euclidean) clustering was used to group similar gene expression profiles, labeled along the right side of each heatmap and listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169670#pone.0169670.s002" target="_blank">S2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169670#pone.0169670.s003" target="_blank">S3</a> Tables. (B) GO categories show relative differences between copper and blue rockfish in the percentage of annotated genes that were differentially expressed, as classified by GO molecular function or biological process. Broken out pie wedges highlight GO categories that were more expressed in one species than the other. Copper rockfish show significant up-regulation of genes involved in transcription and biological regulation at high <i>p</i>CO₂ and down-regulation at low <i>p</i>CO₂. In contrast, blue rockfish differentially express muscle structural genes across <i>p</i>CO₂ treatments.</p

Changes in behavioral lateralization, critical swimming speed, and aerobic scope of juvenile copper and blue rockfish as a function of <i>p</i>CO₂ treatment exposure history.

<p>(A, B) Behavioral lateralization is measured using the relative lateralization index (negative values = right turn bias in a detour test). (C,D) Critical swimming speed (U_crit) is the maximum sustained speed in body lengths per second. (E,F) Aerobic scope represents the difference between maximum and resting metabolic rates (measured as oxygen consumption) and is a proxy for the capacity for aerobic activity. Bars are mean values (± SE). Letters over bars represent results of Tukey HSD post-hoc tests; significantly different means do not share letters in common. Note: Due to logistical constraints all behavioral and physiological trials occurred in control seawater (<i>p</i>CO₂ ~550 μatms).</p

Species-Specific Responses of Juvenile Rockfish to Elevated <i>p</i>CO₂: From Behavior to Genomics

<div><p>In the California Current ecosystem, global climate change is predicted to trigger large-scale changes in ocean chemistry within this century. Ocean acidification—which occurs when increased levels of atmospheric CO₂ dissolve into the ocean—is one of the biggest potential threats to marine life. In a coastal upwelling system, we compared the effects of chronic exposure to low pH (elevated <i>p</i>CO₂) at four treatment levels (i.e., <i>p</i>CO₂ = ambient [500], moderate [750], high [1900], and extreme [2800 μatm]) on behavior, physiology, and patterns of gene expression in white muscle tissue of juvenile rockfish (genus <i>Sebastes</i>), integrating responses from the transcriptome to the whole organism level. Experiments were conducted simultaneously on two closely related species that both inhabit kelp forests, yet differ in early life history traits, to compare high-CO₂ tolerance among species. Our findings indicate that these congeners express different sensitivities to elevated CO₂ levels. Copper rockfish (<i>S</i>. <i>caurinus</i>) exhibited changes in behavioral lateralization, reduced critical swimming speed, depressed aerobic scope, changes in metabolic enzyme activity, and increases in the expression of transcription factors and regulatory genes at high <i>p</i>CO₂ exposure. Blue rockfish (<i>S</i>. <i>mystinus</i>), in contrast, showed no significant changes in behavior, swimming physiology, or aerobic capacity, but did exhibit significant changes in the expression of muscle structural genes as a function of <i>p</i>CO₂, indicating acclimatization potential. The capacity of long-lived, late to mature, commercially important fish to acclimatize and adapt to changing ocean chemistry over the next 50–100 years is likely dependent on species-specific physiological tolerances.</p></div