Summary
A deletion mutant of Rhizobium leguminosarum biovar viciae lacking the host-specific nodulation (nod) gene region (nodFEL nodMNT and nodO) but retaining the other nod genes (nodD nodABCIJ) was unable to nodulate peas or Vicia hirsuta, although it did induce root hair deformation. The mutant appeared to be blocked in its ability to induce infection threads and could be rescued for nodulation of V. hirsuta in mixed inoculation experiments with an exopolysaccharide deficient mutant (which is also Nod−). The nodulation deficiency of the deletion mutant strain could be partially restored by plasmids carrying the nodFE, nodFEL or nodFELMNT genes but not by nodLMN. Surprisingly, the mutant strain could also be complemented with a plasmid that did not carry any of the nodFELMNT genes but which did carry the nodO gene on a 30 kb cloned region of DNA. Using appropriate mutations it was established that nodO is essential for nodulation in the absence of nodFE. Thus, either of two independent nod gene regions can complement the deletion mutant for nodulation of V. hirsuta. Similar observations were made for pea nodulation except that nodL was required in addition to nodO for nodulation in the absence of the nodFE genes. These observations show that nodulation can occur via either of two pathways encoded by non-homologous genes.
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References
Beringer JE (1974) R-factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198
Beynon JL, Beringer JE, Johnston AWB (1980) Plasmids and host range in Rhizobium leguminosarum and Rhizobium phaseoli. J Gen Microbiol 120:413–420
Bibb MJ, Biro S, Motamedi H, Collins JF, Hutchinson CR (1989) Analysis of the nucleotide sequence of the Streptomyces glaucescens tcmI genes provides key information about the enzymology of polyketide antibiotic biosynthesis. EMBO J 8:2727–2736
Bhuvaneswari TV, Solheim B (1985) Root hair deformation in the white clover/Rhizobium trifolii symbiosis. Physiol Plant 63:25–34
Borthakur D, Barber CE, Lamb JW, Daniels MJ, Downie JA, Johnston AWB (1986) A mutation that blocks exopolysaccharide synthesis prevents nodulation of peas by Rhizobium leguminosarum but not of beans by R. phaseoli and is corrected by cloned DNA from Rhizobium or the phytopathogen Xanthomonas. Mol Gen Genet 203:320–323
de Maagd RA, Wijfjes AHM, Spaink HP, Ruiz-Sainz JE, Wijffelman CA, Okker RJH, Lugtenberg BJJ (1989) nodO, a new nod gene of the Rhizobium leguminosarum biovar var. viciae sym plasmid pRLIJI encodes a secreted protein. J Bacteriol 171:6764–6770
Djordjevic MA, Schofield PR, Rolfe BG (1985) Tn5 mutagenesis of Rhizobium trifolii host-specific nodulation genes result in mutants with altered host-range ability. Mol Gen Genet 200:463–471
Djordjevic MA, Innes RW, Wijffelman CA, Schofield PR, Rolfe BG (1986) Nodulation of specific legumes is controlled by several distinct loci in Rhizobium trifolii. Plant Mol Biol 6:389–402
Downie JA (1989) The nodL gene from Rhizobium leguminosarum is homologous to transacetylases encoded by lacA and cysE. Mol Microbiol 3:1649–1651
Downie JA, Johnston AWB (1988) Nodulation of legumes by Rhizobium. Plant Cell Env 11:403–412
Downie JA, Hombrecher G, Ma Q-S, Knight CD, Wells B, Johnston AWB (1983a) Cloned nodulation genes of Rhizobium leguminosarum determine host-range specificity. Mol Gen Genet 190:359–365
Downie JA, Ma QS, Knight CD, Hombrecher G, Johnston AWB (1983b) 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
Downie JA, Knight CD, Johnston AWB, Rossen L (1985) Identification of genes and gene products involved in nodulation of peas by Rhizobium leguminosarum. Mol. Gen Genet 198:255–262
Economou A, Hawkins FKL, Downie JA, Johnston AWB (1989) Transcription of rhiA, a gene on a Rhizobium leguminosarum bv. vicia Sym plasmid, requires rhiR and is repressed by flavanoids that induce nod genes.Mol Microbiol 3:87–93
Economou A, Hamilton WDO, Johnston AWB, Downie JA (1990) The Rhizobium nodulation gene nodO encodes a Ca2+-binding protein that is exported without N-terminal cleavage and is homologous to haemolysin and related proteins. EMBO J 9:349–354
Evans IJ, Downie JA (1986) The nodI gene product of Rhizobium leguminosarum is closely related to the ATP-binding bacterial transport proteins: nucleotide sequence analysis of the nodI and nodJ genes. Gene 43:95–102
Fisher RF, Tu JK, Long SR (1985) Conserved nodulation genes in Rhizobium meliloti and Rhizobium trifolii. Appl Environ Microbiol 49:1439–1435
Honma MA, Ausubel FM (1987) Rhizobium meliloti has three functional copies of the nodD symbiotic regulatory gene. Proc Nail Acad Sci USA 84:8558–8562
Knight CD, Rossen L, Robertson JG, Wells B, Downie JA (1986) Nodulation inhibition by Rhizobium leguminosarum multicopy nodABC genes and analysis of early stages of plant infection. J Bacteriol 166:552–558
Long SR (1989) Rhizobium-legume nodulation: life together in the underground. Cell 56:203–214
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Philip-Hollingsworth S, Hollingsworth RI, Dazzo F (1989) Hostrange related structural features of extracellular polysaccharides of Rhizobium trifolii and Rhizobium leguminosarum. J Biol Chem 264:1461–1466
Rolfe B, Gresshoff P (1988) Genetic analysis of legume nodule initiation. Annu Rev Plant Physiol Plant Mol Biol 39:297–319
Rossen L, Johnston AWB, Downie JA (1984) DNA sequence of the Rhizobium leguminosarum nodulation genes nodAB and C required for root hair curling. Nucleic Acid Res 12:9497–9508
Rostas K, Kondorosi E, Horvath B, Simoncsits A, Kondorosi A (1986) Conservation of extended promoter regions of nodulation genes in Rhizobium. Proc Natl Acad Sci USA 83:1757–1761
Shearman CA, Rossen L, Johnston AWB, Downie JA (1986) The Rhizobium leguminosarum nodulation gene nodF encodes a polypeptide similar to acyl-carrier protein and is regulated by nodD plus a factor in pea root exudate. EMBO J 5:647–652
Sherman DH, Malpartida F, Bibb MJ, Kieser H, Bibb MJ, Hopwood DA (1989) Structure and deduced function of the granaticin-producing cluster of Streptomyces violaceoruber Tü22. EMBO J 8:2717–2725
Spaink HP, Okker RJH, Wijffelman CA, Pees E, Lugtenberg BJJ (1987) Promoters in the nodulation region of the Rhizobium leguminosarum sym plasmid pRl1JI. Plant Mol Biol 9:29–37
Spaink HP, Weinman J, Djordjevic MA, Wijffelman CA, Okker RJH, Lugtenberg BJJ (1989) Genetic analysis and cellular localization of the Rhizobium host-specificity-determining NodE protein. EMBO J 8:2811–2818
Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43
Surin BP, Downie JA (1988) Characterization of the Rhizobium leguminosarum genes nodLMN involved in efficient host specific nodulation. Microbiology 2:173–183
Surin BP, Downie JA (1989) Rhizobium leguminosarum genes required for expression and transfer of host specific nodulation. Plant Mol Biol 12:19–29
Surin BP, Watson JM, Hamilton WDO, Economou A, Downie JA (1990) Molecular characterisation of the nodulation gene nodT, from two biovars of Rhizobium leguminosarum. Mol MicrobioL 4:245–252
Wijffelman CA, Pees E, Van Brussel AAN, Okker RJH, Lugtenberg BJJ (1985) Genetic and functional analysis of the nodulation region of the Rhizobium leguminosarum Sym plasmid pRLIJI. Arch Microbiol 143:225–232
Zaat SAJ, Van Brussel AAN, Tak T, Pees E, Lugtenberg BJ (1987) Flavonoids induce Rhizobium leguminosarum to produce nod-DABC gene-related factors that cause thick, short roots and root hair responses on common vetch. J Bacteriol 169:3388–3391
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Communicated by H. Hennecke
Dedicated to the memory of the late Dr. David Goodchild
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Downie, J.A., Surin, B.P. Either of two nod gene loci can complement the nodulation defect of a nod deletion mutant of Rhizobium leguminosarum bv viciae . Molec. Gen. Genet. 222, 81–86 (1990). https://doi.org/10.1007/BF00283027
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DOI: https://doi.org/10.1007/BF00283027