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Background  5-Methylcytosine (5mC) is an important epigenetic mark in eukaryotes. Little information about its role exists for invertebrates. To investigate the contribution of 5mC to phenotypic variation in invertebrates, alteration of methylation patterns needs to be produced. Here, we apply new non-nucleoside DNA methyltransferase inhibitors (DNMTi) to introduce aleatory changes into the methylome of mollusk species.    Results  Flavanone inhibitor Flv1 was efficient in reducing 5mC in the freshwater snails  Biomphalaria glabrata  and  Physa acuta,  and to a lesser degree, probably due to lower stability in sea water, in the oyster  Crassostrea gigas . Flv1 has no toxic effects and significantly decreased the 5mC level in the treated  B. glabrata  and in its offspring. Drug treatment triggers significant variation in the shell height in both generations. A reduced representation bisulfite-sequencing method called epiGBS corroborates hypomethylation effect of Flv1 in both  B. glabrata  generations and identifies seven Differential Methylated Regions (DMR) out of 32 found both in Flv1-exposed snails and its progeny, from which 5 were hypomethylated, demonstrating a multigenerational effect. By targeted bisulfite sequencing, we confirmed hypomethylation in a  locus  and show that it is associated with reduced gene expression.    Conclusions  Flv1 is a new and efficient DNMTi that can be used to induce transient and heritable modifications of the epigenetic landscape and phenotypic traits in mollusks, a phylum of the invertebrates in which epigenetics is understudied.    Supplementary Information  The online version contains supplementary material available at 10.1186/s13072-021-00422-7.

epigenetics and chromatin

The methylome of Biomphalaria glabrata and other mollusks: enduring modification of epigenetic landscape and phenotypic traits by a new DNA methylation inhibitor