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The Mediterranean sacoglossan  Elysia timida  is one of the few sea slug species with the ability to sequester chloroplasts from its food algae and to subsequently store them in a functional state in the digestive gland cells for more than a month, during which time the plastids retain high photosynthetic activity (= long-term retention). Adult  E. timida  have been described to feed on the unicellular alga  Acetabularia acetabulum  in their natural environment. The suitability of  E. timida  as a laboratory model culture system including its food source was studied.    Results  In contrast to the literature reporting that juvenile  E. timida  feed on  Cladophora dalmatica  first, and later on switch to the adult diet  A. acetabulum , the juveniles in this study fed directly on  A. acetabulum  (young, non-calcified stalks); they did not feed on the various  Cladophora spp.  (collected from the sea or laboratory culture) offered. This could possibly hint to cryptic speciation with no clear morphological differences, but incipient ecological differentiation. Transmission electron microscopy of chloroplasts from  A. acetabulum  after initial intake by juvenile  E. timida  showed different states of degradation — in conglomerations or singularly — and fragments of phagosome membranes, but differed from kleptoplast images of  C. dalmatica  in juvenile  E. timida  from the literature. Based on the finding that the whole life cycle of  E. timida  can be completed with  A. acetabulum  as the sole food source, a laboratory culture system was established. An experiment with PAM-fluorometry showed that cultured  E. timida  are also able to store chloroplasts in long-term retention from  Acetabularia peniculus , which stems from the Indo-Pacific and is not abundant in the natural environment of  E. timida . Variations between three experiment groups indicated potential influences of temperature on photosynthetic capacities.    Conclusions   E. timida  is a viable laboratory model system to study photosynthesis in incorporated chloroplasts (kleptoplasts). Capacities of chloroplast incorporation in  E. timida  were investigated in a closed laboratory culture system with two different chloroplast donors and over extended time periods about threefold longer than previously reported.

frontiers in zoology

Chloroplast incorporation and long-term photosynthetic performance through the life cycle in laboratory cultures of Elysia timida (Sacoglossa, Heterobranchia)