HM was carried out using options available in SVS v.8.8.3 (Golden Helix Inc.). Clusters of ROHs, defined as genomic regions of at least 100 Kb whose loci occur in ROHs of at least ten cultivars characterized at the phenotypic level, were identified. Repeated binary spectral clustering62 (link) was used to trim boundaries of clusters of ROHs, in order to define homozygous regions highly overlapping among cultivars. Finally, a linear regression model was fit between clusters of ROHs and BLUPs, using the top five principal components as covariates to correct for population structure. The FDR correction was used to account for multiple testing and suggest an association for P < 0.1. BLUP means of cultivars either contributing or not contributing to clusters of ROHs associated with phenotypic traits were computed and compared using a two-tail Student’s t test. Genes included in clusters of ROHs identified by HM were retrieved by the Prunus dulcis (cv. Lauranne) v1.0 genome annotation available at the genomic database of Rosaceae28 (link),63 (link).
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