| Abstract: | Beijerinckia indica subsp. indica is an aerobic, acidophilic, exopolysaccharide-producing, N2-fixing soil bacterium. It is a generalist chemoorganotroph that is phylogenetically closely related to facultative and obligate methanotrophs of the genera Methylocella and Methylocapsa. Here we report the full genome sequence of this bacterium.Beijerinckia indica subsp. indica ATCC 9039 is the type strain of the genus Beijerinckia (6), a member of the Rhizobiales order of the Alphaproteobacteria. Beijerinckia spp. are commonly found as free-living bacteria in acidic soils and also in plant rhizosphere and phyllosphere environments (4). Research on Beijerinckia has suffered from chronic taxonomic confusion, with some strains of Sphingomonas and Azotobacter being misidentified in the literature: e.g., a “Beijerinckia” reported to degrade PAH has been reclassified (3). However, some Beijerinckia spp. have received research attention due to their plant growth-promoting properties (7) and for their abundant production of exoheteropolysaccharide with potential biotechnological uses (5).Genomic DNA from Beijerinckia indica subsp. indica was used to create 3-kb, 8-kb, and 40-kb DNA libraries. Sequencing, assembly, and automated annotation were performed at the Joint Genome Institute using standard procedures (U.S. Department of Energy; http://www.jgi.doe.gov/sequencing/strategy.html). The total number of paired-end shotgun Sanger reads in the assembly was 33,870. In addition, Roche 454 sequence data were included into the final assembly. Large Newbler contigs of 454 reads were chopped into 4,975 overlapping fragments of 1,000 bp and entered into the assembly as pseudoreads.The genome of B. indica subsp. indica was 4,170,153 bp. In addition, two plasmids of 181,736 and 66,727 bp were present. There are a total of 3,982 open reading frames (ORFs) predicted using Glimmer, of which 3,784 are predicted protein-coding genes and 2,695 (70%) have been assigned a predicted function. There are 134 pseudogenes, 52 tRNA genes, and three operons each containing 16S, 23S, and 5S rRNA genes. The G+C content is 57.0% (56% and 54% in the plasmids).The bacterium lacks phosphofructokinase, the key enzyme of the Embden-Meyerhof pathway. Instead, it uses the Entner-Doudoroff or pentose phosphate pathway to catabolize sugars, which is typical of free-living Rhizobiales. The majority of the genes involved in N2 fixation are clustered in two genomic islands (10 kb and 51 kb), with the notable exception of the nifS gene encoding cysteine desulfurase.Beijerinckia indica is a metabolically versatile bacterium capable of growth on a variety of organic acids, sugars, and alcohols (4). In contrast, its close phylogenetic cousins Methylocella and Methylocapsa are highly specialized methanotrophs capable of growth on very few substrates (2). However, the genome size of Beijerinckia indica compared to that of Methylocella silvestris (4.17 versus 4.30 Mbp) and the numbers of predicted protein-encoding genes (3,788 versus 3,917) are remarkably similar. A BLAST analysis indicated that the 57% of the genes in the genome of B. indica have homologues in M. silvestris (stringency threshold expectation value [E] of 1e−50). Some key pathways of one-carbon metabolism (such as the tetrahydromethanopterin and serine pathways of formaldehyde metabolism) that are present in M. silvestris appear to be absent or incomplete in B. indica, which confirms previous experiments showing that the organism is incapable of methylotrophic growth (1). However, an operon encoding a putative propane monooxygenase homologous to soluble propane/methane monooxygenases of Methylocella silvestris BL2 was identified. More in-depth comparison of these genomes will help elucidate what defines their distinct lifestyles. |
