Abstract
Elysia crispata is a Sacoglossan sea slug which sequesters, maintains, and utilizes chloroplasts from a wide range of Ulvophycean algae. While many sacoglossan species feed on a small number of algal species, E. crispata is capable of utilizing at least 30 ulvophycean algal species throughout the Caribbean, although individual populations vary tremendously in actual algal chloroplast sources. Most studies have relied on PCR based DNA barcoding techniques combined with plasmid cloning and screening to separate and sequence chloroplast encoded genes (usually rbcL or tufA). However, PCR/cloning-based studies are time consuming, expensive, and may not amplify the targeted gene from all algal species evenly. Recent cost declines in high throughput genome sequencing provides a cost-effective mechanism to examine sequestered chloroplast identities on a much larger scale with more comprehensive coverage. In this note we analyze total genomic DNA from a single E. crispata specimen which was sequenced on the Illumina Hi-Seq platform. Sequences were BLAST searched, filtered, and sorted using OmicsBox v2.1.10 software. Chloroplasts from four algal species could be identified, but over 95% of the rbcL sequences were from just one algal species, Penicillus lamourouxii. The results suggest that the animal either preferred this algal species or that it was the most readily available for consumption.
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Acknowledgements
Sequencing of the slug genome was funded by a University of Tampa RISE grant awarded to Middlebrooks and Mahadevan. Additional support was provided to Curtis by the Dr. Paula Ines Castagnet Endowed Chair of Biological Sciences Award and Ave Maria University. Specimens were collected under pemit SAL-20-0616-SR issued to Middlebrooks by the State of Florida Fish and Wildlife Conservation Commission.
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Curtis, N.E., Middlebrooks, M.M., Mahadevan, P. et al. A methodological note on using next generation sequencing technology to identify the algal sources of stolen chloroplasts in a single sea slug specimen (Elysia crispata) to provide a comprehensive view of the animal’s kleptoplast population. Symbiosis 89, 251–258 (2023). https://doi.org/10.1007/s13199-023-00895-y
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DOI: https://doi.org/10.1007/s13199-023-00895-y