In vivo and in vitro differences in chloroplast functionality in the two north Atlantic sacoglossans (Gastropoda, Opisthobranchia) Placida dendritica and Elysia viridis

The photosynthetic functionality in chloroplasts in the two sacoglossan molluscs Placida dendritica and Elysia viridis from the Trondheim fjord in Norway was studied. P. dendritica and E. viridis with no functional chloroplasts in their digestive system were introduced to the green macroalgae Codium fragile. Our results showed that P. dendritica was not able to retain functional (photosynthetic) chloroplasts. Transmission electron microscopy (TEM) showed that chloroplasts were directly digested when phagocytosed into the digestive cells. Four stages of chloroplast degradation were observed. A corresponding operational quantum yield of chl a fluorescence (I broken vertical bar(PSII) similar to 0) indicated autofluorescence, and the presence of highly degraded chl a supported these observations. In contrast, E. viridis was able to retain functional chloroplasts. For this species it took only 1 week for the chloroplasts inside the digestive cells to acquire the same I broken vertical bar(PSII) and light utilisation coefficient (alpha) as C. fragile kept under the same light conditions. Data for 8 days showed a 2-6-fold increase in the maximum photosynthetic rate (P (max)) and light saturation index (E (k)) relative to C. fragile. This increase in available light was probably caused by a reduced package effect in the digestive gland of E. viridis relative to C. fragile, resulting in a partial photoacclimation response by reducing the turnover time of electrons (tau). Isolated pigments from C. fragile compared to E. viridis showed the same levels of photosynthetic pigments (chl a and b, neoxanthin, violaxanthin, siphonaxanthin, siphonein and beta,epsilon-carotene) relative to mu g chl a (w:w), indicating that the chloroplasts in E. viridis did not synthesise any new pigments. After 73 days of starvation, it was estimated that chloroplasts in E. viridis were able to stay photosynthetic 5-9 months relative to the size of the slugs, corresponding to an RFC of level 8 (a retention ability to retain functional chloroplasts (RFC) for more than 3 months). The reduction in I broken vertical bar(PSII), P (max) and alpha as a function of time was caused by a reduction in chloroplast health and number (chloroplast thylakoid membranes and PSII are degraded). These observations therefore conclude that chloroplasts from C. fragile cannot divide or synthesise new pigments when retained by E. viridis, but are able to partially photoacclimate by decreasing tau as a response to more light. This study also points to the importance of siphonaxanthin and siphonein as chemotaxonomic markers for the identification of algal sources of functional chloroplasts.