Foxtail millet, common millet and Japanese barnyard millet have traditionally been important food sources in East Asian countries. Although waxy types of foxtail and common millet have been identified, a waxy mutant of the allohexaploid crop Japanese barnyard millet has not yet been reported. However, several Japanese landraces have been identified that have approximately half the level of amylose found in other varieties. We employed one of the low amylose landraces, 'Noge-Hie', to produce waxy Japanese barnyard millet using a c-radiation treatment. The seeds from a single M 2 plant stained red-brown with iodine solution, indicating the starch in the seeds lacked amylose. Colorimetric tests indicated that amylose was not present in endosperm tissue of the mutant, and analysis of starch granule bound proteins showed that waxy (Wx) protein was absent from the starch granules. The waxy trait was stably inherited in subsequent generations. Additionally, a PCR-based analysis demonstrated the presence of three separate waxy genes in the millet, and indicated that the low amylose landraces carry a deletion in one of these three genes.