Leaf rust (caused by Puccinia triticina Erikss., Pt) is a severe foliar disease of cultivated wheat worldwide. Severe development of the disease results in significant losses in seed yield and quality. Growing immune varieties is the most rational method for Pt control in terms of effectiveness and ecological safety. However, the gene pool of cultivated wheat is very narrow for seedling Pt effective resistance genes, which hampers breeding for this trait. One of the well-known methods to broaden genetic diversity for resistance is the introgression of highly effective genes from wild relatives into the genomes of cultivated wheat. The Aegilops L. species have been proven to be perfectly suited for this purpose. No gene for Pt resistance has been transferred to wheat from Aegilops biuncialis Vis. (Lorent’s goatgrass) up to now. Previously, we selected eight accessions of the species from the VIR (N.I. Vavilov All-Russian Institute of Plant Genetic Resources) genebank that showed a perfect level of resistance to leaf rust. In this research, we studied the genetic control of resistance using hybridological, phytopathological, and molecular analyses. According to the F1–F3 hybrid evaluation results, each accession possesses one dominant gene for Pt resistance, and genes in different accessions are allelic or very tightly linked. Phytopathological test clone analysis showed that this gene is not identical to Lr9, Lr19, Lr24, Lr39, and Lr47, which are effective against Pt populations in some areas of Russia. This conclusion was partially supported by the results of the identification of DNA markers specific to these genes in bread wheat. Thus, we identified one dominant gene (temporarily symbolized as LrBi1) for effective seedling Pt resistance; it is recommended for introgression to cultivated wheat via interspecific hybridization.