Summary
Symbiotic and auxotrophic mutants of Rhizobium japonicum strain USDA191 were isolated using Tn5 mutagenesis and techniques that cause plasmid deletions and plasmid curing. Characterization of several mutants that are unable to nodulate (Nod-) or unable to fix nitrogen (Fix_) showed that nod and nif genes are located within one regions of a 200 MD plasmid (pSym191). Blot hybridization analysis of plasmids in other fast-growing R. japonicum strains showed that nod as well as nif sequences are located on plasmids in eight strains but are apparently carried in the chromosome in two strains.
Similar content being viewed by others
References
Adams TH, McClung CR, Chelm BK (1984) Physical organization of the Bradyhizobium japonicum nitrogenase gene region. J Bacteriol 159:857–862
Appelbaum E, Kramer R (1980) Restriction mapping of deletions in the nif region of the Klebsiella pneumoniae chromosome. Mol Gen Genet 179:349–354
Appelbaum E, Johansen E, Chartrain N (1984) Identification of plasmids carrying symbiotic genes in fast-growing R. japonicum using DNA hybridization and Tn5 mutagenesis. In: Veeger C, Newton WE (eds) Advances in nitrogen fixation research. Martinus Nijhoff, The Hague, p. 670
Appelbaum E, McLoughlin T, O'Connell M. Chartrain N (1985) Expression of symbiotic genes of Rhizobium japonicum USDA 191 in other rhizobia. J Bacteriol 163:385–388
Beringer JE, Beynon JL, Buchanan-Wollaston AV, Johnston AWB (1978) Transfer of the drug-resistance transposon Tn5 to Rhizobium. Nature 276:633–634
Bishop PE, Guevara JG, Engelke JA, Evans HJ (1976) Relation between glutamine synthetase and nitrogenase activities in the symbiotic association between Rhizobium japonicum and Glycine max. Plant Physiol 57:542–546
Boyer HB, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Brewen NJ, Beringer JB, Johnston AWB (1980) Plasmid-mediated transfer of host-range specificity between two strains of Rhizobium leguminosarum. J Gen Microbiol 120:413–420
Broughton WJ, Heycke N, Meyer H, Pankhurst CE (1984) Plasmid-linked nif and “nod” genes in fast-growing rhizobia that nodulate Glycine max, Psophocarpus tetragonolobus, and Vigna unguiculata. Proc Natl Acad Sci USA 81:3093–3097
Burkardt B, Burkardt H-J (1984) Visualization and exact molecular weight determination of a Rhizobium meliloti megaplasmid. J Mol Biol 175:213–218
Chua KY, Pankhurst CE, MacDonald PE, Hupcroft DH, Jarvis BDW, Scott DB (1985) Isolation and characterization of transposon Tn5-induced symbiotic mutants of Rhizobium loti. J Bacteriol 162:335–343
Cutting JA, Schulman HM (1969) The site of heme synthesis in root nodules. Biochim Biophys Acta 192:486–493
Denarie J, Boistard P, Casse-Delbart F, Atherly AG, Berry JO, Russel P (1981) Indigenous plasmids of Rhizobium. In: Giles KG, Atherly AG (eds) Biology of the Rhizobiaceae. Academic Press, New York, pp 225–264
Downie JA, Ma Q-S, Knight CD, Hombrecher G, Johnston AWB (1983) Cloning of the symbiotic region of Rhizobium leguminosarum: The nodulation genes are between the nitrogenase genes and a nifA-like gene. EMBO J 2:947–952
Eckhardt T (1978) A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid 1:584–588
Egelhoff TT, Fisher RF, Jacobs TW, Mulligan JT, Long SR (1985) Nucleotide sequence of Rhizobium meliloti 1021 nodulation genes: nodD is read divergently from nodABC. DNA 4:241–248
Fuhrmann M, Fischer H-M, Hennecke H (1985) Mapping of Rhizobium japonicum nifB-, fixBC-, and fixA-like genes and identification of the fixA promoter. Mol Gen Genet 199:315–322
Hardy RWF, Holsten RD, Jackson EK, Burns RC (1968) The acetylene-ethylene assay for N2 fixation: Laboratory and field evaluation. Plant Physiol 43:1185–1207
Hattori J, Johnson DA (1984) Fast-growing Rhizobium japonicum that effectively nodulates several commercial Glycine max L. Merrill cultivars. Appl Environ Microbiol 48:234–235
Heron DS, Pueppke SG (1984) Mode of infection,nodulation specificity, and indigenous plasmids of 11 fast-growing Rhizobium japonicum strains. J Bacteriol 160:1061–1066
Hirsch PR (1978) Ph. D. Thesis. University of East Anglia
Holliday R (1956) A new method for the identification of biochemical mutants of microorganisms. Nature 178:987
Hontelez JGJ, Mol P, van Dun C, Schetgens R, van Kammen A, van den Bos RC (1984) Expression of sym-plasmid genes in bacteroids of Rhizobium leguminosarum. In: Veeger C, Newton WE (eds) Advanced in nitrogen fixation research. Nijhoff/Junk Publishers, The Hague p 686
Jorgensen RA, Rothstein SJ, Reznikoff WS (1979) A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet 177:65–72
Keyser HH, Bohlool BB, Hu TS, Weber DF (1982) Fast-growing rhizobia isolated from root nodules of soybean. Science 215:1631–1632
Long SR, Buikema WJ, Ausubel FM (1982) Cloning of Rhizobium meliloti nodulation genes by direct complementation of Nodmutants. Nature 298:485–488
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Masterson RV, Russell PR, Atherly AG (1982) Nitrogen fixation (nif) genes and large plasmids of Rhizobium japonicum. J Bacteriol 152:928–931
Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Morrison NA, Hau CY, Trinick MJ, Shine J, Rolfe BG (1983) Heat curing of a sym plasmid in a fast-growing Rhizobium sp. that is able to nodulate legumes and the nonlegume Parasponia sp. J Bacteriol 153:527–53
Norel F, Kush A, Denefle P, Charpin N, Elmerich C (1984) Nitrogen fixation in a tropical Rhizobium associated with Sesbania rostrata In: Veeger C, Newton WE (eds) Advances in nitrogen fixation research. Nijhoff/Junk Publishers, The Hague, p 694
Prakash RK, Atherly AG (1984) Reiteration of genes involved in symbiotic nitrogen fixation by fast-growing Rhizobium japonicum. J Bacteriol 160:785–787
Riedel GE, Ausubel FM, Cannon FC (1979) Physical map of chromosomal nitrogen fixation (nif) genes of Klebsiella pneumoniae. Proc Natl Acad Sci USA 76:2866–2870
Rosenberg C, Casse-Delbart F, Dusha I, David M, Boucher C (1982) Megaplasmids in the plant-associated bacteria Rhizobium meliloti and Pseudomonas solanacearum. J Bacteriol 150:402–406
Rostas K, Sista P, Stanley J, Verma DP (1984) Transposon mutagenesis of Rhizobium japonicum. Mol Gen Genet 197:230–235
Rossen L, Ma Q-S, Mudd EA, Johnston AWB, Downie JA (1984) Identification and DNA sequence of fixZ, a nifB-like gene from Rhizobium leguminosarum. Nucl Acids Res 12:7123–7134
Ruvkun GB, Ausubel FM (1980) Interspecies homology of nitrogenase genes. Proc Natl Acad Sci USA 77:191–195
Sadowsky MJ, Bohlool BB (1983) Possible involvement of a megaplasmid in nodulation of soybeans by fast-growing rhizobia from China. Appl Environ Microbiol 46:906–911
Scott KF, Rolfe BG, Shine J (1981) Biological nitrogen fixation: primary structure of the Klebsiella pneumoniae nifH and nifD genes. J Mol Appl Genet 1:71–81
Scott DB, Court CB, Ronson CW, Scott KF, Watson JM, Schofield PR, Shine J (1984) Organization of nodulation and nitrogen fixation genes on a Rhizobium trifolii symbiotic plasmid. Arch Microbiol 139:151–157
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 range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Biotechnology 1:784–791
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Szeto WW, Zimmerman JL, Sundaresan V, Ausubel FM (1984) A Rhizobium meliloti symbiotic regulatory gene. Cell 36:1035–1043
Veeger C, Newton WE (1984) Advances in nitrogen fixation research. Martinus Nijhoff, The Hague
Vincent JM (1970) A manual for the practical study of root-nodule bacterial. Blackwell Scientific Publications. Oxford
Yelton MM, Yang SS, Eadie SA, Lim ST (1983) Characterization of an effective salt-tolerant, fast-growing strain of Rhizobium japonicum. J Gen Microbiol 129:1537–1547
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Appelbaum, E.R., Johansen, E. & Chartrain, N. Symbiotic mutants of USDA191, a fast-growing Rhizobium that nodulates soybeans. Molec Gen Genet 201, 454–461 (1985). https://doi.org/10.1007/BF00331339
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331339