Abstract
The dinoflagellate Symbiodiniaceae family plays a central role in the health of the coral reef ecosystem via the symbiosis that establishes with its inhabiting cnidarians and supports the host metabolism. In the last few decades, coral reefs have been threatened by pollution and rising temperatures which have led to coral loss. These events have raised interest in studying Symbiodiniaceae and their hosts; however, progress in understanding their metabolism, signal transduction pathways, and physiology in general, has been slow because dinoflagellates present peculiar characteristics. We took advantage of one of these peculiarities; namely, the post-transcriptional addition of a Dino Spliced Leader (Dino-SL) to the 5′ end of the nuclear mRNAs, and used it to generate cDNA libraries from Symbiodinium microadriaticum. We compared sequences from two Yeast-Two Hybrid System cDNA Libraries, one based on the Dino-SL sequence, and the other based on the SMART technology (Switching Mechanism at 5′ end of RNA Transcript) which exploits the template switching function of the reverse transcriptase. Upon comparison of the performance of both libraries, we obtained a significantly higher yield, number and length of sequences, number of transcripts, and better 5′ representation from the Dino-SL based library than from the SMART library. In addition, we confirmed that the cDNAs from the Dino-SL library were adequately expressed in the yeast cells used for the Yeast-Two Hybrid System which resulted in successful screening for putative SmicRACK1 ligands, which yielded a putative hemerythrin-like protein.
Highlights
Symbiodiniaceae are highly diverse dinoflagellate algae that establish symbiosis with cnidarians and marine organisms of diverse phyla including corals
Coral reefs have suffered large areal losses due to coral death caused by diseases and coral bleaching, the latter being a cause of the symbiosis breakdown; the mechanisms underlying the metabolism, symbiont selection and establishment, maintenance, and disruption of the symbiosis, are still poorly understood
Because the cDNA content quality could affect the performance of the libraries, we determined and compared the yield of each library to confirm differences in their complexity
Summary
Symbiodiniaceae are highly diverse dinoflagellate algae that establish symbiosis with cnidarians and marine organisms of diverse phyla including corals. Coral reefs shelter a highly biodiverse ecosystem, which depends on the functional symbiosis with dinoflagellate members of such a family [1,2]. The dinoflagellates reside in symbiosomal membranes in the host gastrodermal cells, where they carry out the photosynthetic process that fuels the productivity and diversity of the coral reef ecosystems. Microorganisms 2021, 9, 791 cnidarian–dinoflagellate symbiosis is a consequence of the scarcely available molecular and functional genomic tools applied to corals and Symbiodiniaceae. It has not been possible to obtain silenced, mutated, or knockout stable lines of Symbiodiniaceae to carry out integral functional genomic studies that may shed light on these pathways [3,4]
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