Banana black leaf streak, caused by Mycosphaerella fijiensis M. Morelet, is a primary phytosanitary concern in export of this fruit around the world. To develop improved cultivars resistant to this disease, an understanding of host response to infection is necessary. In this study, we obtained expression data on 14,872 genes by microarray analysis in the resistant genotype Musa acuminata subsp. burmannicoides 'Calcutta 4' after inoculation with Mycosphaerella fijiensis. From these data, 16 genes were analyzed as potential reference genes and 12 genes were identified as potential early indicators of the onset of the host defense response. Subsequently, these genes were analyzed by quantitative reverse-transcription polymerase chain reaction in susceptible 'Williams' and resistant Calcutta 4. The 18S and 26S ribosomal subunit genes in both cultivars showed the best characteristics as reference genes. In all, 5 of the 12 defense genes expressed shortly after infection (peroxidase, pathogenesis-related [PR]-4, PR-10, phenylalanine ammonia-liase, and disease resistance response 1) showed overexpression in Calcutta 4 between 6 and 24 h after inoculation as opposed to Williams, which did not show overexpression after 144 h. Early induction of defense-related genes could be a key component of the resistance of the Calcutta 4 genotype against M. fijiensis. In addition, these five genes could be used as indicators of the activation of defense responses in the interaction between banana and M. fijiensis.