Rhizobia are symbiotic nitrogen-fixing soil bacteria that are associated with host legumes. The establishment of rhizobial symbiosis requires signal exchanges between partners in microaerobic environments that result in mutualism for the two partners. We developed a macroarray for Mesorhizobium loti MAFF303099, a microsymbiont of the model legume Lotus japonicus, and monitored the transcriptional dynamics of the bacterium during symbiosis, microaerobiosis, and starvation. Global transcriptional profiling demonstrated that the clusters of genes within the symbiosis island (611 kb), a transmissible region distinct from other chromosomal regions, are collectively expressed during symbiosis, whereas genes outside the island are downregulated. This finding implies that the huge symbiosis island functions as clustered expression islands to support symbiotic nitrogen fixation. Interestingly, most transposase genes on the symbiosis island were highly upregulated in bacteroids, as were nif, fix, fdx, and rpoN. The genome region containing the fixNOPQ genes outside the symbiosis island was markedly upregulated as another expression island under both microaerobic and symbiotic conditions. The symbiosis profiling data suggested that there was activation of amino acid metabolism, as well as nif-fix gene expression. In contrast, genes for cell wall synthesis, cell division, DNA replication, and flagella were strongly repressed in differentiated bacteroids. A highly upregulated gene in bacteroids, mlr5932 (encoding 1-aminocyclopropane-1-carboxylate deaminase), was disrupted and was confirmed to be involved in nodulation enhancement, indicating that disruption of highly expressed genes is a useful strategy for exploring novel gene functions in symbiosis.Through the symbiotic nitrogen fixation process, bacteria belonging to the family Rhizobiaceae convert atmospheric dinitrogen (N 2 ) to ammonia (NH 3 ), which can be effectively used by host legume plants. The establishment of a rhizobiumlegume symbiosis requires induction of new developmental programs in the partners. The symbiotic interaction begins with signal exchanges of flavonoids and Nod factors (lipochitooligosaccharides) between the two partners (6). In legume nodules, microaerobic environments trigger the rhizobial expression of nitrogen-fixing genes, such as nif and fix, via an oxygen-sensing system (13). However, the establishment of nitrogen-fixing symbiosis probably requires more complex steps triggered by reciprocal signal exchanges that lead to the organogenesis of nodules, differentiation of microsymbionts, and efficacy of nodulation (27). In addition to this symbiotic lifestyle, rhizobia survive in soils with many environment stresses, such as nutrient starvation.Lotus japonicus is a promising model legume for studying molecular interactions between symbiosis partners (20). Schauser et al. (40) first identified the plant regulatory gene nin, which is responsible for the nodule organogenesis program, in this legume. Recently, the receptor-like kinase genes have...