During meiosis in flowering plants, degradation of the callose wall in tetrads releases newly produced microspores, which develop into mature pollen grains. In this study, we identified zbs1, a male-sterile mutant of naked oat (Avena nuda L.) that displayed complete spikelet sterility due to inviable mature pollen. The abnormal pollen grains originated from microspores with a defective callose wall and cell plate during meiosis. The defective callose wall and cell plate of the zbs1 mutant were detected by the labeling of cell wall epitopes (β-1,3-glucan) with immunogold during meiosis, and an abnormal chromosome configuration was observed by propidium iodide staining. The mature pollen grains of the zbs1 mutant were irregular in shape, and abnormal germination was observed by scanning electron microscopy. Together, our results indicate that the cause of male sterility in zbs1 is abnormal meiosis.