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Pyrosequencing-based transcriptomic resources in the pond snail Lymnaea stagnalis, with a focus on genes involved in molecular response to diquat-induced stress

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Abstract

Due to their ability to explore whole genome response to drugs and stressors, omics-based approaches are widely used in toxicology and ecotoxicology, and identified as powerful tools for future ecological risk assessment and environmental monitoring programs. Understanding the long-term effects of contaminants may indeed benefit from the coupling of genomics and eco-evolutionary hypotheses. Next-generation sequencing provides a new way to investigate pollutants impact, by targeting early responses, screening chemicals, and directly quantifying gene expression, even in organisms without reference genome. Lymnaea stagnalis is a freshwater mollusk in which access to genomic resources is critical for many scientific issues, especially in ecotoxicology. We used 454-pyrosequencing to obtain new transcriptomic resources in L. stagnalis and to preliminarily explore gene expression response to a redox-cycling pesticide, diquat. We obtained 151,967 and 128,945 high-quality reads from control and diquat-exposed individuals, respectively. Sequence assembly provided 141,999 contigs, of which 124,387 were singletons. BlastX search revealed significant match for 34.6 % of the contigs (21.2 % protein hits). KEGG annotation showed a predominance of hits with genes involved in energy metabolism and circulatory system, and revealed more than 400 putative genes involved in oxidative stress, cellular/molecular stress and signaling pathways, apoptosis, and metabolism of xenobiotics. Results also suggest that diquat may have a great diversity of molecular effects. Moreover, new genetic markers (putative SNPs) were discovered. We also created a Ensembl-like web-tool for data-mining (http://genotoul-contigbrowser.toulouse.inra.fr:9095/Lymnaea_stagnalis/index.html). This resource is expected to be relevant for any genomic approach aimed at understanding the molecular basis of physiological and evolutionary responses to environmental stress in L. stagnalis.

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Acknowledgments

The project has been carried out with INRA financial support (EFPA Projet Innovant 2010; AIP Bioressources 2010, project EGELYS). AB is granted by the INRA (PhD, Contrat Jeune Scientifique). The authors thank Christophe Plomion for advices and insightful comments on the project and for critical reading of the manuscript, and Marc Collinet and the INRA U3E staff for technical assistance.

Ethical standards

The authors declare that the experiments were performed in compliance with current french laws.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. INRA, UMR0985 INRA-Agrocampus Ouest ESE, Equipe Ecotoxicologie et Qualité des Milieux Aquatiques, 65 rue de Saint-Brieuc, 35042, Rennes cedex, France

    Anthony Bouétard, Anne-Laure Besnard, Damien Choisne, Laurent Lagadic & Marie-Agnès Coutellec

  2. Genotoul, Plateforme Bioinformatique, INRA Toulouse-Auzeville, Chemin de Borderouge, 31326, Castanet Tolosan, France

    Céline Noirot & Christophe Klopp

  3. Genotoul, Plateforme Génomique, INRA UMR0444 LGC Toulouse-Auzeville, Chemin de Borderouge, 31326, Castanet Tolosan, France

    Olivier Bouchez & Eugénie Robe

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  1. Anthony Bouétard
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Correspondence to Marie-Agnès Coutellec.

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Anthony Bouétard and Céline Noirot contributed equally to this work.

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Bouétard, A., Noirot, C., Besnard, AL. et al. Pyrosequencing-based transcriptomic resources in the pond snail Lymnaea stagnalis, with a focus on genes involved in molecular response to diquat-induced stress. Ecotoxicology 21, 2222–2234 (2012). https://doi.org/10.1007/s10646-012-0977-1

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