Fine mapping and functional characterization of SiLAC3 in regulating brown seed coat pigmentation, integrated...

The color of the seed coat in sesame is a critical breeding trait, with significant implications for seed quality and consumer preferences. However, the genetic foundation and regulatory mechanisms underlying seed coat colors remain largely unexplored. Earlier, we identified a quantitative trait locus (QTL), qBSCchr6, associated with brown-colored seed coats in sesame. Currently, we utilized a recombinant inbred line (RIL) G3010, which harbors qBSCchr6 and crossed it with the white-seeded cultivar (YZ8) to develop an F₂ population for QTL fine mapping. Through detailed recombinant analysis, qBSCchr6 was fine mapped to a narrow 83.4 kb genomic interval, co-segregating with the molecular marker BSC_SNV23. Within this interval, we identified SIN_1023239 for the brown-colored seed coat, which we have designated as SiLAC3. SiLAC3 encodes a laccase enzyme with three putative multicopper oxidase domains essential for its function. Sequence analysis revealed that a single nucleotide variant in the multicopper oxidase_3 domain is critical for the expression of brown seed coat phenotype. To further elucidate the role of SiLAC3, we employed a combination of chemical staining, transcriptomics, and metabolomics analyses. Our results indicate that the lignin biosynthesis pathway is crucial for brown seed coat pigmentation, with SiLAC3 facilitating oxidative polymerization of coniferyl alcohol in the seed coat. These findings advance our understanding of genetic and molecular mechanisms underlying seed coat color in sesame and offer a foundation to improve this trait in breeding programs.