Two closely linked novel loci, qKRN2-1 and qKRN2-2, associated with kernel row number were fine-mapped on chromosome 2, and a key candidate gene for qKRN2-1 was identified through expression analysis. Kernel row number (KRN) is a crucial factor influencing maize yield and serves as a significant target for maize breeding. The use of wild progenitor species can aid in identifying the essential traits for domestication and breeding. In this study, teosinte (MT1) served as the donor parent, the inbred maize line of Mo17 was used as the recurrent parent, we identified a major quantitative trait locus (QTL) for KRN, designated qKRN2, into two closely linked loci, qKRN2-1 and qKRN2-2. Here, fine mapping was performed to investigate two QTLs, qKRN2-1 and qKRN2-2, within a genomic range of 272kb and 775kb, respectively. This was achieved using a progeny test strategy in an advanced backcross population, with the two QTLs explaining 33.49% and 35.30% of the phenotypic variance. Molecular marker-assisted selection resulted in the development of two nearly isogenic lines (NILs), qKRN2-1 and qKRN2-2, which differed only in the segment containing the QTL. Notably, the maize (Mo17) alleles increased the KRN relative to teosinte by approximately 1.4 and 1.2 rows for qKRN2-1 and qKRN2-2, respectively. Zm00001d002989 encodes a cytokinin oxidase/dehydrogenase and its expression in the immature ears exhibited significant differences among the qKRN2-1 NILs. In situ hybridization localized Zm00001d002989 to the primordia of the inflorescence meristem and spikelet pair meristems, is predicted to be the causal gene of qKRN2-1. The findings of this study deepen our understanding of the genetic basis of KRN and hold significant potential for improving maize grain yields.
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