Recombination within the Cepaea nemoralis supergene is confounded by incomplete penetrance and epistasis

Although the land snail Cepaea nemoralis is one of the most thoroughly investigated colour polymorphic species, there have been few recent studies on the inheritance of the shell traits. Previously, it has been shown that the shell polymorphism is controlled by a series of nine or more loci, of which five make a single ‘supergene’ containing tightly linked colour and banding loci and more loosely linked pigmentation, spread band and punctate loci. However, one limitation of earlier work was that putative instances of recombination between loci within the supergene were not easily verified. We therefore generated a new set of C. nemoralis crosses that segregate for colour, banding and pigmentation, and several other unlinked shell phenotype loci. The snails were genotyped using a set of RAD-seq loci that flank the supergene, and instances of recombination tested by comparing inferred supergene genotype against RAD-marker genotype. We found no evidence that suspected ‘recombinant’ individuals are recombinant within the supergene - point estimates of recombination between both colour/banding, and colour/pigmentation loci are zero, with upper limits of 0.8 and 1.8%. Incomplete penetrance and epistasis are a better explanation for the apparent ‘recombinant’ phenotypes. Overall, this work therefore provides a resource for fine mapping of the supergene and other major shell phenotype loci. It also shows that the architecture of the supergene may not be as previously supposed.