The marine slug Elysia chlorotica (Gould) forms an intracellular symbiosis with photosynthetically active chloroplasts from the chromophytic alga Vaucheria litorea (C. Agardh). This symbiotic association was characterized over a period of 8 months during which E. chlorotica was deprived of V. litorea but provided with light and CO2. The fine structure of the symbiotic chloroplasts remained intact in E. chlorotica even after 8 months of starvation as revealed by electron microscopy. Southern blot analysis of total DNA from E. chlorotica indicated that algal genes, i.e., rbcL, rbcS, psaB, psbA, and 16S rRNA are present in the animal. These genes are typically localized to the plastid genome in higher plants and algae except rbcS, which is nuclear-encoded in higher plants and green (chlorophyll a/b) algae. Our analysis suggests, however, that similar to the few other chromophytes (chlorophyll a/c) examined, rbcS is chloroplast encoded in V. litorea. Levels of psbA transcripts remained constant in E. chlorotica starved for 2 and 3 months and then gradually declined over the next 5 months corresponding with senescence of the animal in culture and in nature. The RNA synthesis inhibitor 6-methylpurine reduced the accumulation of psbA transcripts confirming active transcription. In contrast to psbA, levels of 16S rRNA transcripts remained constant throughout the starvation period. The levels of the photosystem II proteins, Dl and CP43, were high at 2 and 4 months of starvation and remained constant at a lower steady-state level after 6 months. In contrast, D2 protein levels, although high at 2 and 4 months, were very low at all other periods of starvation. At 8 months, de novo synthesis of several thylakoid membrane-enriched proteins, including Dl, still occurred. To our knowledge, these results represent the first molecular evidence for active transcription and translation of algal chloroplast genes in an animal host and are discussed in relation to the endosymbiotic theory of eukaryote origins.The ascoglossan sea slug Elysia chlorotica (Gould) forms an intracellular symbiosis with chloroplasts of the filamentous chromophytic alga Vaucheria litorea (C. Agardh) (1, 2). The animal resembles a dark green leaf and is capable of photoautotrophic CO2 fixation as a result of the high density of chloroplasts dispersed throughout its extensive digestive system (1, 3, 4). The photosynthetic sea slug survives in laboratory aquaria for 8-9 months when provided with only light and CO2 (starved); a time period similar to its life cycle in nature. The symbiotic association is not inherited since the plastids are not transmitted in the eggs (5). Instead, chloroplast symbiosis is reestablished with each new generation of sea slugs (1, 6, 7). Acquisition of chloroplast symbionts begins immediately following metamorphosis from the veliger stage when juvenile sea slugs begin to feed on V litorea cells (1, 2). Once ingested, the chloroplasts are phagocytically incorporated into the cytoplasm of one of two morphologically distinct...