The SigD-regulated genes in Bacillus subtilis were systematically analyzed by comparing the pattern of transcripts derived from wild-type cells with those from sigD mutant cells using DNA microarray technology. One hundred and fifty-eight genes were found to be SigD-dependent candidates, 46 of which being known SigD-regulated genes. Northern blot analysis revealed that 18 of the remaining genes were SigD-dependent. The SigD consensus sequence was newly identified in the upstream regions of nine operons (11 genes): ybdO, yfmT-yfmS, hemAT, yjcP-yjcQ, yjfB, ylqB, yoaH, yscB and yxkC, and the other seven genes were assumed to be indirectly affected by a SigD mutation. Furthermore, yviE-yviF are likely to be SigD-dependent genes, because three independent sets of array data for yviE and yviF indicated they are SigD-dependent, and these genes are neighbors of flgL and hag transcribed by SigD RNA polymerase.