1  

The following supplement accompanies the article

Plasticity and trade–offs in physiological traits of intertidal mussels when confronting freshwater–induced environmental variation

Laura Ramajo, Luis Prado, Alejandro B. Rodriguez–Navarro, Marco A. Lardies, Carlos M. Duarte, Nelson A. Lagos*

*Corresponding author: nlagoss@ust.cl

Marine Ecology Progress Series 535: 93–109 (2016)

Table S1. Perumytilus purpuratus. Mean (± Standard Error) in maximum shell size, wet and buoyant weight of mussels used in the reciprocal transplant experiment (n = 60 mussels per treatment).

Origin Destination Treatment Shell sizea (mm) Wet Weightb (g) Buoyant Weightc (g)

Marine Site Marine Site Auto-transplant 13.27 (0.54) 0.6277 (0.0890) 0.2288 (0.0329)

River-Infl. Site Transplant 13.29 (0.54) 0.6715 (0.1000) 0.2312 (0.0339)

River-Infl. Site River-Infl. Site Auto-transplant 13.41 (0.44) 0.5468 (0.0495) 0.2002 (0.0190)

Marine Site Transplant 13.28 (0.43) 0.5399 (0.0520) 0.1989 (0.0195) One-way ANOVA: aF1, 240 = 0.02, p = 0.895. bF1, 240 = 2.02, p = 0.156. cF1, 240 = 1.29, p = 0.258.

2  

Table S2. Effects on growth, calcification and metabolic rates in Perumytilus purpuratus individuals depending on origin (Las Cruces and San Antonio) and destination site (auto-transplanted and transplant treatment) after 56 days. In bold are showed significant p-values at α ≤ 0.05.

Biological responses Source DF MS F P-value

Growth Rate (mm d-1)

Origin Site (OS) 1 0.0002 0.13 0.724 Destination Site (DS) 1 0.0021 1.71 0.193 OS × DS 1 0.0293 24.22 <0.001 Error 164 0.0012 Total 167

Net Calcification Rate (mgCaCO3 g-1 d-1)

Origin Site (OS) 1 0.0002 0.52 0.471 Destination Site (DS) 1 0.0007 1.48 0.226 OS × DS 1 0.0113 22.7 <0.001 Error 169 0.0005 Total 172

3  

Figure S1. Image of a Perumytilus purpuratus individual labeled with a bee tag (A) and photography of the experimental cages used in the experiment (B).

(A)

(B)

4  

Figure S2. Perumytilus purpuratus. (A) Growth rate (mm d-1) and (B) net calcification rate (mgCaCO3g-1 d-1) of Perumytilus purpuratus cross-transplanted between Las Cruces and San Antonio localities at 56 d of beginning of the experiment. Data are means ± SE. Different letters beside each symbol indicate significant differences between experimental treatments evaluated using Tukey HSD test as post hoc comparison.

5  

Figure S3. SEM analyses of fractured shell samples (a) and periostracum (b) of Perumytilus purpuratus.

6  

Figure S4. Discriminant functions analysis generated using relative levels (absorbance values from FTIR) of carbonate signals (CO3) in combination with (A) proteins and (B) lipids recorded on the shell periostracum of Perumytilus purpuratus for each treatment after 154 days of cross-reciprocal experiment. Results of multivariate analysis of variance (MANOVA) are shown.