Abstract:Kernel-related traits, including kernel length, kernel width, and hundred kernel weight, are pivotal factors in determining maize yield. In this study, a recombinant inbred line (RIL) population derived from the maize inbred lines B73 and CML277 was utilized as the experimental material. Based on the high-precision genetic maps constructed through genotyping-by-sequencing (GBS) technology, nine kernel-related quantitative trait loci (QTL) were identified. The major locus qKW2.04 was precisely located on chromosome 2 using the inclusive composite interval mapping (ICIM) method, explaining 20.34% and 15.84% of the phenotypic variation on grain width and 100-grain weight, respectively. In addition, an F2 segregating population was developed using the recurrent parent B73 and its introgression line (NIL-1041A) to further delimit qKW2.04. This locus was found with contribution due to two tightly linked kernel width major QTL, designated as qKW2.04-1 and qKW2.04-2, which were located in the marker intervals InDel23.32~umc1555 and InDel47.09~InDel57.06, explaining 22.45% and 12.22% of the phenotypic variation, respectively. The favorable allelic variations for both QTLs were contributed by CML277. Through the selection of recombinant individuals within the target region, qKW2.04-1 was finely positioned between molecular markers InDel26.76~InDel27.86 within a 1.1 Mb interval. This study provides new clues for elucidating the genetic basis of kernel-related traits and gene resources for high-yield molecular breeding in maize.