Ultraviolet screening by slug tissue and tight packing of plastids protect photosynthetic sea slugs from photoinhibition.

One of the main mysteries regarding photosynthetic sea slugs is how the slug plastids handle photoinhibition, the constant light-induced damage to Photosystem II of photosynthesis. Recovery from photoinhibition involves proteins encoded by both the nuclear and plastid genomes, and slugs with plastids isolated from the algal nucleus are therefore expected to be incapable of constantly repairing the damage as the plastids inside the slugs grow old. We studied photoinhibition-related properties of the sea slug Elysia timida that ingests its plastids from the green alga Acetabularia acetabulum. Spectral analysis of both the slugs and the algae revealed that there are two ways the slugs use to avoid major photoinhibition of their plastids. Firstly, highly photoinhibitory UV radiation is screened by the slug tissue or mucus before it reaches the plastids. Secondly, the slugs pack the plastids tightly in their thick bodies, and therefore plastids in the outer layers protect the inner ones from photoinhibition. Both properties are expected to greatly improve the longevity of the plastids inside the slugs, as the plastids do not need to repair excessive amounts of damage.

Genetic autonomy and low singlet oxygen yield support kleptoplast functionality in photosynthetic sea slugs.

The kleptoplastic sea slug Elysia chlorotica consumes Vaucheria litorea, stealing its plastids, which then photosynthesize inside the animal cells for months. We investigated the properties of V. litorea plastids to understand how they withstand the rigors of photosynthesis in isolation. Transcription of specific genes in laboratory-isolated V. litorea plastids was monitored for 7 days. The involvement of plastid-encoded FtsH, a key plastid maintenance protease, in recovery from photoinhibition in V. litorea was estimated in cycloheximide-treated cells. In vitro comparison of V. litorea and spinach thylakoids was applied to investigate reactive oxygen species formation in V. litorea. In comparison to other tested genes, the transcripts of ftsH and translation elongation factor EF-Tu (tufA) decreased slowly in isolated V. litorea plastids. Higher levels of FtsH were also evident in cycloheximide-treated cells during recovery from photoinhibition. Charge recombination in PSII of V. litorea was found to be fine-tuned to produce only small quantities of singlet oxygen, and the plastids also contained reactive oxygen species-protective compounds. Our results support the view that the genetic characteristics of the plastids are crucial in creating a photosynthetic sea slug. The plastid's autonomous repair machinery is likely enhanced by low singlet oxygen production and elevated expression of FtsH.

On the art of stealing chloroplasts.

Sea slugs increase the longevity of the chloroplasts they steal from algae by limiting the harmful side-effects of photosynthesis.

Photosynthetic sea slugs induce protective changes to the light reactions of the chloroplasts they steal from algae.

Sacoglossan sea slugs are able to maintain functional chloroplasts inside their own cells, and mechanisms that allow preservation of the chloroplasts are unknown. We found that the slug Elysia timida induces changes to the photosynthetic light reactions of the chloroplasts it steals from the alga Acetabularia acetabulum. Working with a large continuous laboratory culture of both the slugs (>500 individuals) and their prey algae, we show that the plastoquinone pool of slug chloroplasts remains oxidized, which can suppress reactive oxygen species formation. Slug chloroplasts also rapidly build up a strong proton-motive force upon a dark-to-light transition, which helps them to rapidly switch on photoprotective non-photochemical quenching of excitation energy. Finally, our results suggest that chloroplasts inside E. timida rely on oxygen-dependent electron sinks during rapid changes in light intensity. These photoprotective mechanisms are expected to contribute to the long-term functionality of the chloroplasts inside the slugs.

Plants, algae and a few other organisms rely on a process known as photosynthesis to fuel themselves, as they can harness cellular structures called chloroplasts to convert light into usable energy. Animals typically lack chloroplasts, making them unable to use photosynthesis to power themselves. The sea slug Elysia timida, however, can steal whole chloroplasts from the cells of the algae it consumes: the stolen structures then become part of the cells in the gut of the slug, allowing the animal to gain energy from sunlight. Once they are in the digestive system of the slug, the chloroplasts survive and keep working for longer than expected. Indeed, these structures are often harmed as a side effect of photosynthesis, but the sea slug does not have the right genes to help repair this damage. In addition, conditions inside animal cells are widely different to the ones found inside algae and plants. It is not clear then how the sea slug extends the lifespan of its chloroplasts by preventing damage caused by sunlight. To investigate this question, Havurinne and Tyystjärvi compared photosynthesis in sea slugs and the algae they eat. A range of methods, including measuring fluorescence from the chloroplasts, was used: this revealed that the slug changes the inside of the stolen chloroplasts, making them more resistant to damage. First, when exposed to light the stolen chloroplasts can quickly switch on a mechanism that dissipates light energy to heat, which is less damaging. Second, a molecule that serves as an intermediate during photosynthesis is kept in a 'safe' state which prevents it from creating harmful compounds. And finally, additional safeguard molecules 'deactivate' compounds that could otherwise mediate damaging reactions. Overall, these measures may reduce the efficiency of the chloroplasts but allow them to keep working for much longer. Early chloroplasts were probably independent bacteria that were captured and 'domesticated' by other cells for their ability to extract energy from the sun. Photosynthesizing sea slugs therefore provide an interesting way to understand some of the challenges of early life. The work by Havurinne and Tyystjärvi may also reveal new ways to harness biological processes such as photosynthesis for energy production in other contexts.

Why It Is Time to Look Beyond Algal Genes in Photosynthetic Slugs.

Eukaryotic organelles depend on nuclear genes to perpetuate their biochemical integrity. This is true for mitochondria in all eukaryotes and plastids in plants and algae. Then how do kleptoplasts, plastids that are sequestered by some sacoglossan sea slugs, survive in the animals' digestive gland cells in the absence of the algal nucleus encoding the vast majority of organellar proteins? For almost two decades, lateral gene transfer (LGT) from algae to slugs appeared to offer a solution, but RNA-seq analysis, later supported by genome sequencing of slug DNA, failed to find any evidence for such LGT events. Yet, isolated reports continue to be published and are readily discussed by the popular press and social media, making the data on LGT and its support for kleptoplast longevity appear controversial. However, when we take a sober look at the methods used, we realize that caution is warranted in how the results are interpreted. There is no evidence that the evolution of kleptoplasty in sea slugs involves LGT events. Based on what we know about photosystem maintenance in embryophyte plastids, we assume kleptoplasts depend on nuclear genes. However, studies have shown that some isolated algal plastids are, by nature, more robust than those of land plants. The evolution of kleptoplasty in green sea slugs involves many promising and unexplored phenomena, but there is no evidence that any of these require the expression of slug genes of algal origin.