An NGS approach for the identification of precise homoeologous recombination sites between A and C genomes in Brassica genus.

The introgression of heterologous genomes through interspecific hybridization offers a great opportunity to expand the gene pool of crops, thereby broadening the traits that can be targeted for improvement. The introgression of C genomic regions carrying desirable traits from Brassica napus (AACC) into the diploid B. rapa (AA) via homoeologous recombination (HR) has been commonly used. However, the precise identification of HR sites remains a significant challenge, limiting the practical application of genome introgression via HR in breeding programs. Here, we developed an indicator named 'Dosage-score' from the coverage depth of next-generation sequencing reads. Then, Dosage-score analysis applied to both in BC₁F₁ individuals obtained by backcrossing B. rapa to F₁ progeny (B. rapa × B. napus) and in the parental lines, and successfully identified the precise HR sites resulting from F₁ meiosis as well as those that were native in the parental B. napus genome. Additionally, we introgressed the C6 segment from HR identified by Dosage-score analysis into B. rapa genome background, revealing gene expression on the added segment without noticeable phenotypic change. The identification of HR by Dosage-score analysis will contribute to the expansion of the gene pool for breeding by introgression of heterologous genomes in Brassica crops.