Summary
Rhizobium sp. NGR234 in a fast-growing Rhizobium strain with a broad host range. The location and role of chromosomal genes involved in cellular metabolism or in the legume symbioses is unknown. We isolated a series of auxotrophic and antibiotic resistant mutants of NGR234 and utilized a chromosome mobilization system based on Tn5-Mob and pJB3JI; Tn5-Mob donor strains behaved like Hfr strains, transferring the chromosome polarly at high frequency from a fixed point of insertion. The use of four different strains with Tn5-Mob located at different nutritional loci in crosses with double auxotrophic recipients, allowed us to build up a circular linkage map of NGR234 based on relative recombination frequencies. Also, symbiotically important genes identified by site-directed mutagenesis, such as hemA and ntrA, could be located and mapped on the chromosome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Tc:
-
tetracycline
- Sp:
-
spectinomycin
- Rif:
-
rifampicin
- Km:
-
kanamycin
References
Beringer JE (1974) R-factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198
Beringer JE, Hoggan SA, Johnston AWB (1978) Linkage mapping in Rhizobium leguminosarum by means of R plasmid-mediated recombination. J Gen Microbiol 104:201–207
Brewin NJ, Beringer JE, Johnston AWB (1980) Plasmid-mediated transfer of host-range specificity between two strains of Rhizobium leguminosarum. J Gen Microbiol 120:413–420
Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 207–209
Dowling DN, Samrey U, Stanley J, Broughton WJ (1987) Cloning of Rhizobium leguminosarum genes for competitive nodulation blocking on peas. J Bacteriol 169:1345–1348
Finan TM, Oresnik I, Bottacin A (1988) Mutants of Rhizobium meliloti defective in succinate metabolism. J Bacteriol 170:3396–3403
Forrai T, Vincze E, Banfalvi Z, Kiss GB, Randhawa GS, Kondorosi A (1983) Localization of symbiotic mutations in Rhizobium meliloti. J Bacteriol 153:635–643
Holliday R (1956) A new method for the identification of biochemical mutants of micro-organisms. Nature 178:987
Hooykaas PJJ, Peerbolte R, Regensburg-Tuink AJG, de Vries P, Schilperoort RA (1982) A chromosomal linkage map of Agrobacterium tumefaciens and a comparison with the maps of Rhizobium spp. Mol Gen Genet 188:12–17
Kondorosi A, Kiss GB, Forrai T, Vincze E, Benfalvi Z (1977) Circular linkage map of Rhizobium meliloti chromosome. Nature 268:525–527
Kondorosi A, Vincze E, Johnston AWB, Beringer JE (1980) A comparison of three Rhizobium linkage maps. Mol Gen Genet 178:403–408
Leong SA, Ditta GS, Helinski DR (1982) Heme biosynthesis in Rhizobium. J Biol Chem 257:8724–8730
Lewin A, Rosenberg C, Meyer ZAH, Wong CH, Nelson L, Marten J-F, Stanley J, Dowling DN, Denarie J, Broughton WJ (1987) Multiple host-specificity loci of the broad host-range Rhizobium sp. NGR234 selected using the widely compatible legume Vigna unguiculata. Plant Mol Biol 8:447–459
Low KB (1987) Mapping techniques and determination of chromosome size. In: Niedhardt FC (ed) Escherichia coli and Salmonella typhimurium. Cellular and Molecular Biology. American Society for Microbiology, Washington DC, pp 1184–1189
Meade HM, Signer ER (1977) Genetic mapping of Rhizobium meliloti. Proc Natl Acad Sci USA 74:2076–2078
Megias M, Caviedes MA, Palomares AJ, Perez-Silva J (1982) Use of plasmid R68.45 for constructing a circular linkage map of the Rhizobium trifolii chromosome. J Bacteriol 149:59–64
Miller IS, Fox D, Saeed N, Borland PA, Miles CA, Sastry GRK (1986) Enlarged map of Agrobacterium tumefaciens C58 and the location of the chromosomal regions which affect tumorigenicity. Mol Gen Genet 205:153–159
Simon R (1984) High frequency mobilization of Gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet 196:413–420
Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology 1:784–791
Simon R, O'Connell M, Labes M, Pühler A (1986) Plasmid vectors for the genetic analysis and manipulation of Rhizobia and other Gram-negative bacteria. Methods Enzymol 118:640–659
Stanley J, Dowling DN, Broughton WJ (1988) Cloning of hemA of Rhizobium sp. NGR234 and symbiotic phenotype of a gene-directed mutant in diverse legume genera. Mol Gen Genet 215:32–37
Stanley J, van Slooten J, Dowling DN, Finan T, Broughton WJ (1989) Cloning, site-directed mutagenesis and phenotypes of the ntrA gene of the groad host-range Rhizobium sp. NGR234. Mol Gen Genet 217:528–532
Trinick MJ (1980) Relationship amongst the fast growing rhizobia of Lablab purpureus, Leucaena leucocephala, Mimosa sp., Acacia farnesiana and Sesbania grandiflora and their affinities with other rhizobial groups. J Appl Bacteriol 49:39–53
Vincent JM (1970) A handbook for the practical study of rootnodule bacteria. IBP handbook 15. Blackwell, Oxford
Yakobson EA, Guiney DG (1984) Conjugal transfer of bacterial chromosomes mediated by the RK2 plasmid transfer origin cloned into transposon Tn5. J Bacteriol 160:451–453
Author information
Authors and Affiliations
Additional information
Communicated by J. Lengeler
Rights and permissions
About this article
Cite this article
Østerås, M., Stanley, J., Broughton, W.J. et al. A chromosomal genetic map of Rhizobium sp. NGR234 generated with Tn5-Mob. Molec. Gen. Genet. 220, 157–160 (1989). https://doi.org/10.1007/BF00260871
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00260871