Genome-wide association analysis provides molecular insights into the natural variation of watermelon seed size.

Seed-consumption watermelon tend to have larger-sized seeds, while flesh-consumed watermelon often require relatively smaller seed. Therefore, the seed size of watermelon has received extensive attention from consumers and breeders. However, the study on the natural variation and genetic mechanism of watermelon seed size is not clear enough. In the present study, 100 seed weight, seed hilum length, seed length, seed width, and seed thickness in 197 watermelon accessions were examined. Furthermore, association analysis was conducted between seed size traits and high-quality SNP data. The results revealed that there was a strong correlation between the five seed traits. And seed enlargement was an important feature during watermelon seed size domestication. Meanwhile, the seed consumption biological species C. mucosospermu and C. lanatus edible seed watermelon had a significantly bigger seed size than other species's. Eleven non-repeating significant SNPs above the threshold line were obtained by GWAS analysis. Four of them on chromosome 5 were considered to be closely associated with seed size traits, i.e. S5: 32250307, S5: 32250454, S5: 32256177, S5: 32260870, which could be used as potential molecular markers for the breeding of watermelon cultivars with target seed size. In addition, combined with gene annotation information and previous reports, five genes near the four significant SNPs may regulate seed size. And qRT-PCR analysis showed that two genes Cla97C05G104360 and Cla97C05G104380, which may be involved in abscisic acid metabolism, may play an important role in regulating the seed size of watermelon. Our findings provide molecular insights into natural variation in watermelon seed size, and gives valuable information of molecular marker-assisted breeding.