Unveiling protist diversity associated with the Pacific oyster Crassostrea gigas using blocking and excluding primers

Background

Microbiome of macroorganisms might directly or indirectly influence host development and homeostasis. Many studies focused on the diversity and distribution of prokaryotes within these assemblages, but the eukaryotic microbial compartment remains underexplored so far.

Results

To tackle this issue, we compared blocking and excluding primers to analyze microeukaryotic communities associated with Crassostrea gigas oysters. High-throughput sequencing of 18S rRNA genes variable loops revealed that excluding primers performed better by not amplifying oyster DNA, whereas the blocking primer did not totally prevent host contaminations. However, blocking and excluding primers showed similar pattern of alpha and beta diversities when protist communities were sequenced using metabarcoding. Alveolata, Stramenopiles and Archaeplastida were the main protist phyla associated with oysters. In particular, Codonellopsis, Cyclotella, Gymnodinium, Polarella, Trichodina, and Woloszynskia were the dominant genera. The potential pathogen Alexandrium was also found in high abundances within some samples.

Conclusions

Our study revealed the main protist taxa within oysters as well as the occurrence of potential oyster pathogens. These new primer sets are promising tools to better understand oyster homeostasis and disease development, such as the Pacific Oyster Mortality Syndrome (POMS) targeting juveniles.

Keyword(s)

Holobiont, Metabarcoding, Microbiome, Ostreoida, Crassostrea

Full Text

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Publisher's official version
132 Mo
Additional file 1: Table S1. Microbiota samples
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Additional file 2: Figure S1. Sequences of high taxonomic ranks (oysters, Embryophyceae, protists) within oyster sample
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Additional file 3: Table S2. Number of sequences. Values correspond to 18SV1V2EX/18SV4BP.
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Additional file 4: Figure S2. Rarefaction analyses.
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Additional file 5: Table S3. OTU annotations and abundances for 18SV1V2EX samples.
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Additional file 6: Table S4. OTU annotations and abundances for 18SV4BP samples.
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Additional file 7: Figure S3. Comparison of amplicon sizes between 18SV1V2EX and 18SV4BP
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Additional file 8: Table S5. Metadata and alpha diversity indices. Alpha diversity values correspond to 18SV1V2EX/18SV4BP. NA: not analysed (protist sequences< 5160).
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How to cite
Clerissi Camille, Guillou Laure, Escoubas Jean Michel, Toulza Eve (2020). Unveiling protist diversity associated with the Pacific oyster Crassostrea gigas using blocking and excluding primers. Bmc Microbiology. 20 (1). 193 (13p.). https://doi.org/10.1186/s12866-020-01860-1, https://archimer.ifremer.fr/doc/00636/74799/

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