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Wheat provides a substantial portion of globally consumed calories, yet faces growing challenges to meet yield needs. Traits affecting floral architecture and grain number hold immense promise for increasing yield potential. Multi-ovary wheat is a unique mutant line that holds potential in improving yields, characterized by the formation of 2-3 grains per floret in a given wheat spike. Described is a multi-approach mapping effort that refines the chromosomal position of the MOV1 locus at high resolution. A radiation hybrid map, linkage map, and refined physical map are reported. Further mutation analysis recovered missense mutations within a candidate gene, displaying a quantitatively reduced phenotype. Expression profiling of the candidate gene reveals up-regulation at crucial developmental stages. Structural investigation of the gene and surrounding locus identified two insertion/deletion events that suggest a pathway for transcriptional regulation. The results presented implicate the candidate gene in the manifestation of the multi-ovary phenotype.