Heterosis or hybrid vigor describes a phenomenon that superior phenotypes compared to the two parents are observed in the heterozygous F1-hybrid plants. Identification and characterization of heterosis-related genes (HRGs) will facilitate hybrid breeding in crops. To identify HRGs in Brassica rapa, we analyzed transcriptome profiling using a Br300K microarray in non-heading Chinese cabbage at three developmental stages. A large number of genes were differentially expressed in F1 hybrids and non-additive expression was prominent. Genes that are expressed specifically for F1 hybrid at all three stages were Brassica-specific uncharacterized genes and several defense-related genes. Expression of several photosynthesis- and stress-related genes were also F1 hybrid-specific. Thirteen NBS-LRR class genes showed high and specific expression in F1 hybrid Shulu: some of them were characterized as defense genes in Arabidopsis, but most have not been. Further characterization of these defense-related genes in Brassica species and its application will be helpful for understanding the role of defense responses in heterosis. In addition, results obtained in this study will be valuable to develop molecular markers for heterosis and disease resistance in B. rapa.