Rapid Mining of Candidate Genes for the Branchless Phenotype in Watermelon by Bulked Segregant Analysis Using Whole-genome Resequencing

Lateral branching is an important agronomic trait in horticulture plants. The aim of this project was to reveal the genetic mechanism, map the gene localization, and predict candidate genes of watermelon lateral branching. An F2 segregating population was derived from a cross between the multibranched maternal inbred line M6 and the branchless paternal inbred line N7. Two DNA pools were constructed using 20 multibranched plants and 20 branchless plants from the F2 population. Whole-genome resequencing was performed for the DNA pools (25×) and the parents (30×) to identify the genomic region associated with lateral branching. Candidate genes were predicted based on the gene annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses; then, quantitative validation of these genes was performed. The results showed that the clean reads of four samples yielded 64,295,076 to 81,658,958 bp, with sufficient genome coverage and high quality. Based on single-nucleotide polymorphism and insertions/deletions association analyses, the candidate genes were mapped to a 2.01-Mb region on chromosome 4 (22,958,925–24,971,894 bp) containing 182 annotated genes. During the KEGG and GO enrichment analyses, these genes were annotated to 10 cellular components, 10 molecular functions, and 12 biological processes. Eight candidate genes responsible for the branchless phenotype in watermelon were identified: Cla97C04G076340, Cla97C04G075820, Cla97C04G076060, Cla97C04G076250, Cla97C04G076280, Cla97C04G076380, Cla97C04G076830, and Cla97C04G075950. These genes were involved in amino acid biosynthesis and catabolism, TCP transcription factor activity, and regulation of flower development. This study offers valuable insights into the molecular mechanisms governing the branchless phenotype in watermelon. These candidate genes serve as potential targets for gene cloning and marker-assisted selection of watermelon cultivars without lateral branches.