Smc03964 is predicted to possess a twin-arginine export signal [64], and to encode a member of the metallophosphatase superfamily (cl13995), a group of phosphatases with diverse functions [52]. ORFs SMc01424, SMc01423, and SMc01422 appear to be part of a single operon and they encode, respectively,
a predicted nitrile hydratase alpha subunit protein, a nitrile hydratase beta subunit protein, and a nitrile hydratase activator protein [53, 54]. Nitrile hydratases function in the degradation of xenobiotic compounds, but they are also involved in tryptophan metabolism, specifically in the #selleck screening library randurls[1|1|,|CHEM1|]# conversion of 3-indoleacetonitrile to indole-3-acetamide, which is a precursor of the plant hormone auxin [65, 66]. SMa0044 has an unusual expression pattern in that it is expressed at a very low level in approximately half of the nodules tested (Table 3; Figure 4), but is expressed quite strongly by free-living S. meliloti on LBMC medium ( Additional file 5). SMa0044 is predicted to encode a member of the DUF2277 superfamily, which is has no known function [52]. Conclusions The goal 4-Hydroxytamoxifen cell line of this study was to identify S. meliloti 1021 ORFs involved in host plant nodulation and nitrogen fixation. The comparative genomics method
we employed was able to rediscover 19 ORFs that have previously been shown to be important for nodulation and/or nitrogen fixation. The earlier studies that identified these genes, in most cases, employed the classical bacterial genetic techniques of transposon mutagenesis, followed by strain isolation and phenotypic screening [11, 67][68]. Our study identified 9 additional S. meliloti ORFs (out of the 13 we analyzed) that we have shown are expressed primarily in host plant nodules. Thiamine-diphosphate kinase However none of these newly identified ORFs were required for development of a functional symbiosis under the conditions we tested. Our results suggest that the accumulated transposon screens
for essential S. meliloti nodulation/nitrogen fixation genes may be nearing saturation. However, the comparative genomics method described above might be very effective for identifying factors involved in the production of a phenotype common to a group of bacterial species that have not yet been studied by classical transposon mutagenesis screens. Acknowledgments The authors wish to thank Sharon Long, Melanie Barnett, and Jeanne Harris for plasmid pJH104; Graham Walker for plasmid pK19mobsac; and Michiko E. Taga, Penny J. Beuning and George W. Bates for critical reading of the manuscript. This work was funded by start-up funds provided to KMJ by Florida State University. Electronic supplementary material Additional file 1 : Table S1. Joint Genome Institute, Integrated Microbial Genomes Phylogenetic Profile search data on single genes. (XLS 102 KB) Additional file 2 : Table S2. Primers used to amplify S. meliloti 1021 fragments for construction of insertion mutants and deletion mutants. (XLS 54 KB) Additional file 3 : Table S3.