Bivalve transcriptomics reveal pathogen sequences and a powerful immune response of the Mediterranean mussel (Mytilus galloprovincialis)
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Bivalves have colonized the interface between land and sea for the last 500 million years. Although bivalves lack an adaptive immune system, they are extraordinarily well adapted to adverse environmental conditions. Bivalves are valuable aquaculture resources worldwide and are used as sentinels for monitoring pollution. In this work, the immune transcriptomes of mussels (Mytilus galloprovincialis and edulis) and clam (Ruditapes decussatus) were sequenced. For comparative purposes, an already published transcriptome dataset of Ruditapes philippinarum was also included in the analyses. The 454 pyrosequencing of stimulated hemocytes resulted in more than 400,000 reads for each transcriptome. The percentage of annotated sequences ranges from 50% for mussels to 30–40% for clams. Considering the 28,061 non-redundant sequences from the four transcriptomes, the four species share 785 genes. Moreover, sequences related to different putative pathogens were found in the four bivalves. A high number of bivalve herpesvirus ORFs were found, which confirms the value of NGSs as tools to detect and quantify pathogen RNA. Based on an examination of the immune-enriched transcriptomes of these four species, we can conclude that bivalves present an immune system that differs from its conventional characterization as a simple innate immune response against invading pathogens. Enrichment analyses showed that species in the Mytilus genus, especially M. galloprovincialis, possesses a significantly higher number of sequences related to immune processes and killing molecules than species in the Ruditapes genus. This could be related to the broader ecological niche occupied by mussels and the scarcity of reported mussel mass mortalities compared to the high number of mass mortalities reported for clams.
This work has been funded by the EU Project REPROSEED (245119) and 10 PXIB 402 096 PR from Xunta de Galicia; and partially supported by Ministerio de Economía y Competitividad through Intramural 201640E024 and MYTIPEP (AGL2015-65705-R). We also acknowledge the support of Xunta de Galicia to our group (IN607B 2016/12). RM wishes to acknowledge the Spanish MICINN for her FPI Spanish research Grant (BES-2009-029765) and the EU H2020 funded Project VIVALDI (678589). We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). We would also like to thank Ainhoa Blanco for the supply of the blue mussel, Mytilus edulis.
BN, LB and AF conceived and designed the project. PB and MM performed the annotation step. RM and PB made the functional annotation analyses. GFC did the BUSCO analyses. BN and RM made the analysis of pathogen sequences. RM wrote the manuscript. All listed authors revised, edited, read and approved the manuscript.
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Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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