In flowering plants, the tapetum is proposed to play a vital role in the early stages of pollen development. Disruptions to tapetum development and degeneration typically result in male sterility. The present study characterised a maize (Zea mays L.) anther-specific gene, Zm401, which only contains short open reading frames (sORFs). The longest ORF of the Zm401 gene encodes a small protein designated Zm401p10 that accumulates in the nucleus. Overexpression of Zm401p10 in maize retarded tapetal degeneration and caused microspore abnormalities. A microarray analysis identified 278 downregulated and 150 upregulated genes in anthers overexpressing Zm401p10. These results indicate that the Zm401 gene is one of the major components of the molecular network regulating maize anther development and male fertility, and that Zm401p10 is expressed from the longest ORF of the gene.