The r-X1 deficiency in maize produces high frequencies of aneuploid progeny by inducing nondisjunction during embryo sac development. The purpose of this investigation was to determine the embryo sac division at which the nondisjunctive event takes place. Monosomic-6 individuals were generated by crossing r-X1 containing plants by those carrying the y-pastel-8549 allele of the Y locus on chromosome 6. A strict dosage relationship exists between the number of dominant alleles of the Y locus and the level of β-carotene in the endosperm. We found that the level of β-carotene in the endosperm of kernels containing monosomic-6 embryos was comparable with that found in endosperms with one Y allele. This indicated that they contained only one maternally contributed chromosome 6. Only nondisjunction at the second postmeiotic division could have produced such an endosperm; thus, the nondisjunctive event occurs at this division. Another endosperm type expected from nondisjunction at this same division, with three dominant Y alleles, was not found. The absence of this endosperm type suggests that the egg nucleus and the polar nucleus originating at the micropylar pole are sister nuclei that arose from the same cell at the third embryo sac mitotic division. Key words: aneuploidy, embryo sac, nondisjunction, r-X1 deficiency, Zea.