Fine mapping and identification of the leaf shape gene BoFL in ornamental kale

BoFL, a candidate gene conferring the feathered-leaved trait in ornamental kale, is located in a 374.532-kb region on chromosome C9. Leaf shape is an important economic trait in ornamental kale (Brassica oleracea var. acephala). Identifying the genes that influence leaf shape would provide insight into the mechanism underlying leaf development. In this study, we constructed F₁, F₂, BC₁P₁, BC₁P₂, and F_2:3 populations from a cross between a feathered-leaved inbred line, F0819, and a smooth-leaved inbred line, S0835, of ornamental kale. Genetic analysis showed that the feathered-leaved trait was controlled by a semi-dominant gene, BoFL. Using bulked segregant analysis sequencing, we mapped the BoFL gene to a 4.17-Mb interval on chromosome C9. Next, we narrowed down the candidate region to 374.532-kb by fine-scale mapping between the two flanking markers INDEL940 and INDEL5443. We identified 38 genes in the candidate region, among which only Bo9g184610 showed significant variation in expression level between the two parental lines. Sequencing of the gene in the parental lines identified three single-nucleotide polymorphism (SNP) differences, containing two nonsynonymous and one synonymous SNPs, which resulted in coding variations of an aspartate and a phenylalanine residue in F0819, but an alanine and a leucine residue in S0835. A cleaved amplified polymorphic sequence (CAPS) marker, CAPS4610, corresponding to the first nonsynonymous SNP co-segregated with the leaf shape trait. We thus speculated that the gene conferring the feathered-leaved trait is BoALG10, a homolog of ALG10, which encodes an alpha-1,2-glucosyltransferase in Arabidopsis thaliana. This work will be useful for understanding the mechanism of leaf development and provides important information for the breeding of kale with novel leaf shapes.