TY - JOUR T1 - A Genome-Wide Association Analysis Reveals Epistatic Cancellation of Additive Genetic Variance for Root Length in <em>Arabidopsis thaliana</em> JF - bioRxiv DO - 10.1101/008789 SP - 008789 AU - Jennifer Lachowiec AU - Xia Shen AU - Christine Queitsch AU - Örjan Carlborg Y1 - 2015/01/01 UR - http://biorxiv.org/content/early/2015/06/10/008789.abstract N2 - Efforts to identify loci underlying complex traits generally assume that most genetic variance is additive. Here, we examined the genetics of Arabidopsis thaliana root length and found that the narrow-sense heritability for this trait was statistically zero. This low additive genetic variance likely explains why no associations to root length could be found using standard additive-model-based genome-wide association (GWA) approaches. However, the broad-sense heritability for root length was significantly larger, and we therefore also performed an epistatic GWA analysis to map loci contributing to the epistatic genetic variance. This analysis revealed four interacting pairs involving seven chromosomal loci that passed a standard multiple-testing corrected significance threshold. Explorations of the genotype-phenotype maps for these pairs revealed that the detected epistasis cancelled out the additive genetic variance, explaining why these loci were not detected in the additive GWA analysis. Small population sizes, such as in our experiment, increase the risk of identifying false epistatic interactions due to testing for associations with very large numbers of multi-marker genotypes in few phenotyped individuals. Therefore, we estimated the false-positive risk using a new statistical approach that suggested half of the associated pairs to be true positive associations. Our experimental evaluation of candidate genes within the seven associated loci suggests that this estimate is conservative; we identified functional candidate genes that affected root development in four loci that were part of three of the pairs. In summary, statistical epistatic analyses were found to be indispensable for confirming known, and identifying several new, functional candidate genes for root length using a population of wild-collected A. thaliana accessions. We also illustrated how epistatic cancellation of the additive genetic variance resulted in an insignificant narrow-sense, but significant broad-sense heritability that could be dissected into the contributions of several individual loci using a combination of careful statistical epistatic analyses and functional genetic experiments.Author summary Complex traits, such as many human diseases or climate adaptation and production traits in crops, arise through the action and interaction of many genes and environmental factors. Classic approaches to identify contributing genes generally assume that these factors contribute mainly additive genetic variance. Recent methods, such as genome-wide association studies, often adhere to this additive genetics paradigm. However, additive models of complex traits do not reflect that genes can also contribute with non-additive genetic variance. In this study, we use Arabidopsis thaliana to determine the additive and non-additive genetic contributions to the phenotypic variation in root length. Surprisingly, much of the observed phenotypic variation in root length across genetically divergent strains was explained by epistasis. We mapped seven loci contributing to the epistatic genetic variance and validated four genes in these loci with mutant analysis. For three of these genes, this is their first implication in root development. Together, our results emphasize the importance of considering both non-additive and additive genetic variance when dissecting complex trait variation, in order not to lose sensitivity in genetic analyses. ER -