We analysed the products of male meiosis in microspore-derived progeny from a Brassica napus (AACⁿCⁿ) × Brassica carinata (BBC^cC^c) interspecific hybrid (ABCⁿC^c). Genotyping at 102 microsatellite marker loci and nuclear DNA contents provided strong evidence that 26 of the 28 progeny (93%) were derived from unreduced (2n) gametes. The high level of CⁿC^c marker heterozygosity, and parallel spindles at Anaphase II in the ABCⁿC^c hybrid, indicated that unreduced gametes were formed by first division restitution. The frequency of dyads at the tetrad stage of pollen development (2.6%) suggested that unreduced gametes were preferentially selected in microspore culture. Segregation of marker alleles in the microspore-derived progeny was consistent with homologous recombination between Cⁿ and C^c chromosomes and homoeologous recombination involving A-, B- and C-genome chromosomes during meiosis in the ABCⁿC^c hybrid. We discuss the potential for using microspore culture of unreduced gametes in interspecific hybrids to map Brassica centromeres through half-tetrad analysis.