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Volume 143, Issue 6

The symbiotic phenotypes of exopolysaccharide-defective mutants of sp. strain TAL1145 do not differ on determinate- and indeterminate-nodulating tree legumes Free

Abstract

Three classes of exopolysaccharide (EPS) defective mutants were isolated by Tn -insertion mutagenesis of sp. strain TAL1145, which nodulates tree legumes. The class I and class III mutants produced 10-22% of the EPS produced by TAL1145 and appeared partially mucoid while the class II mutants formed small, opaque and non-mucoid colonies. Size-fractionation of the soluble EPSs made by these mutants in the culture supernatant indicated that the class I and the class III mutants produced reduced levels of both highland low-molecular-mass EPSs while the class II mutants lacked both these EPSs but produced a small amount of a medium-molecular-mass anthrone-reactive EPS. The succinyl and acetyl substituents observed in the TAL1145 EPS were absent in the EPS of the class II mutants. When examined under UV, the class I and class III mutants grown on Calcofluor-containing YEM agar showed dim blue fluorescence, compared to the bright blue fluorescence of the wild-type strain, whereas the class II mutants did not fluoresce. While the dim blue fluorescence of the class III mutants changed to yellow-green after 10 d, the fluorescence of the class I mutants did not change after prolonged incubation. Unlike the EPS-defective mutants of other rhizobia, these mutants did not show different symbiotic phenotypes on determinate- and indeterminate-nodulating tree legumes. The class I and the class III mutants formed small ineffective nodules on both types of legumes whereas the class II mutants formed normal nitrogen-fixing nodules on both types. The genes disrupted in the class I and class III mutants form a single complementation group while those disrupted in the class II mutants constitute another. All the three classes of EPS-defective mutants were located within a 10.8 kb region and complemented by two overlapping cosmids.

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Keyword(s): exo genes , nitrogen fixation and nodulation
© Society Society for General Microbiology 1997
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1997-06-01
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References

  1. Borthakur D., Barber C.E., Lamb J.W., Daniels M.J., Downie J.A., Johnston A. W. B. 1986; A mutation that blocks exopolysaccharide synthesis prevents nodulation of peas by Rhizobium leguminosarum but not of beans by Rhizobium phaseol i and is corrected by cloned DNA from Rhizobium or the phytopathogen Xanthomonas.. Mol Gen Genet 203:320–323
     [Google Scholar]
  2. Borthakur D., Barber C.E., Latchford J.W., Rossen L., Johnston A. W. B. 1988; Analysis of pss genes of Rhizobium leguminosarum required for exopolysaccharide synthesis and nodulation of peas : their primary structure and their interaction with psi and other nodulation genes.. Mol Gen Genet 213:155–162
     [Google Scholar]
  3. Chen H., Batley M., Redmond J., Rolfe B.G. 1988; Alteration of the effective nodulation properties of a fast-growing broad host range Rhizobium due to changes in exopolysaccharide synthesis. J Plant Physiol 120:331–349
     [Google Scholar]
  4. Corzo H., Perez-Galdona R., Leon-Barrios M., Gutierrez- Navarro, A.M. 1988; Alcian blue fixation allows staining of the isolated polysaccharide component of bacterial lipopolysaccharides in polyacrylamide gels. Electrophoresis 12:439–441
     [Google Scholar]
  5. Diebold R., Noel K.D. 1989; Rhizobium kguminosarum exopolysaccharide mutants : biochemical and genetic analyses and symbiotic behavior on three hosts.. J Bacteriol 171:4821–4830
     [Google Scholar]
  6. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X.-W., Finlay D.R., Guiney D., Helinski D.R. 1985; Plasmids related to the broad host-range vector, pRK290, useful for gene cloning and monitoring gene expression.. Plasmid 13:149–153
     [Google Scholar]
  7. Figurski D.H., Helinski D.R. 1979; Replication of an origincontaining derivative of plasmid RK2 dependent on a plasmid function provided in trans.. Proc Natl Acad Sci U S A 76:1648–1652
     [Google Scholar]
  8. George M. L. C., Young J. P. W., Borthakur D. 1994; Genetic characterization of Rhizobium sp. strain TALll45 that nodulates tree legumes.. Can J Microbiol 40:208–215
     [Google Scholar]
  9. Glazebrook J., Walker G.C. 1989; A novel polysaccharide can function in place of the Calcofluor-binding exopolysaccharide in nodulation of alfalfa by Rhizobium meliloti .. Cell 56:661–672
     [Google Scholar]
  10. Gray J.X., Zhan H., Levery S.B., Battisti L., Rolfe B.G., Leigh, J.A. 1991; Heterologous exopolysaccharide production in Rhizobium sp . strain NGR234 and consequences for nodule development.. J Bacteriol 173:3066–3077
     [Google Scholar]
  11. Hollingsworth R.I., Dazzo F.B. 1988; The complete structure of the trifoliin : a lectin-binding capsular polysaccharide of Rhizobium trifolii 843. . Carbohydr Res 172:97–112
     [Google Scholar]
  12. Hotter G.S., Scott D.B. 1991; Exopolysaccharide mutants of Rhizobium loti are fully effective on a determinate nodulating host but are ineffective on an indeterminate nodulating host.. J Bacteriol 173:851–859
     [Google Scholar]
  13. Jefferson R.A., Kavanagh T.A., Bevan M.W. 1987; GUS fusions : β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.. EMBO J 6:3901–3907
     [Google Scholar]
  14. Kim C.-H., Tully R.E., Keister D.L. 1989; Exopolysaccharidedeficient mutants of Rhizobium fredii HH303 which are symbiotically effective.. Appl Environ Microbiol 55:1852–1854
     [Google Scholar]
  15. KO Y.-H., Gadya R. 1990; Nodule formation in soybeans by exopolysaccharide mutants of Rhizobium fredii USDA191.. J Gen Microbiol 136:105–113
     [Google Scholar]
  16. Leigh J.A., Coplin D.L. 1992; Exopolysaccharides in plantbacterial interactions.. Annu Rev Microbiol 46:305–346
     [Google Scholar]
  17. Leigh J.A., Lee D.L. 1988; Characterization of polysaccharides of Rhizobium meliloti exo mutants that form ineffective nodules.. J Bacteriol 170:3327–3332
     [Google Scholar]
  18. Leigh J.A., Walker G.C. 1994; Exopolysaccharides of Rhizobium : synthesis, regulation and symbiotic function.. Trends Genet 10:63–67
     [Google Scholar]
  19. Leigh J.A., Signer E.R., Walker G.C. 1985; Exopolysaccharide- deficient mutants of Rhizobium meliloti that form ineffective nodules.. Proc Natl Acad Sci USA 82:6231–6234
     [Google Scholar]
  20. Leigh J.A., Reed J.W.., Hanke J.F., Hirsch A.M., Walker G.C. 1987; Rhizobium meliloti mutants that fail to succinylate their calcofluor-binding exopolysaccharide are defective in nodule invasion. . Cell 51:579–587
     [Google Scholar]
  21. Long S., Reed J.W., Himawan J., Walker G.C. 1987; Genetic analysis of a cluster of genes required for the synthesis of the calcofluor-binding exopolysaccharide of Rhizobium mefiloti .. J Bacteriol 170:4239–4248
     [Google Scholar]
  22. Lopez-Lara I.M., Orgambide G., Dazzo F.B., Olivarez J., Toro N. 1993; Characterization and symbiotic importance of acidic extracellular polysaccharides of Rhizobium sp. strain GRH2 isolated from Acacia nodules.. J Bacteriol 10:2826–2832
     [Google Scholar]
  23. McNeil M., Darvill J., Darvill A.G., Albersheim P. 1986; The discernible structural features of the acidic polysaccharides secreted by different Rhizobium species are the same.. Carbohydr Res 146:307–326
     [Google Scholar]
  24. Parniske M., Schmidt P.E., Kosch K., Muller P. 1994; Plant defense responses of host plants with determinate nodules induced by EPS-defective exoB mutants of Bradyrhizobium japonicum .. Mol Plant-Microbe Interact 7:631–638
     [Google Scholar]
  25. Parveen N., Borthakur D. 1994; Construction of a singletransposon- insertion mutant in Rhizobium sp. strain TALl145 from a double-insertion mutant.. Lett Appl Microbiol 19:142–145
     [Google Scholar]
  26. Parveen N., Webb D.T., Borthakur D. 1996; Leucaena leucocephala nodules formed by a surface polysaccharide defective mutant of Rhizobium sp. strain TAL1145 are delayed in bacteroid development and nitrogen fixation.. Mol Plant-Microbe Interact 9:364–372
     [Google Scholar]
  27. Pooyan S., George M.L.C., Borthakur D. 1996; Isolation and characterization of a gene for nodule development linked to the ndvA and ndvB genes in Rhizobium sp. strain TAL1145.. Symbiosis 17:201–215
     [Google Scholar]
  28. Reuhs B.L., Williams M.N.V., Kim J.S., Carlson R. W., Cote, F. 1995; Suppression of the Fix- phenotype of Rhizobium meliloti exoB mutants by lpsZ is correlated to a modified expression of K polysaccharide.. J Bacteriol 177:489–4296
     [Google Scholar]
  29. Ruvkun G.B., Ausubel F.M. 1981; A general method for sitedirected mutagenesis in prokaryotes.. Nature 289:85–88
     [Google Scholar]
  30. Soedarjo M., Hemscheidt T.K., Borthakur D. 1994; Mimosime, a toxin present in leguminous trees ( Leucaena spp.) induces a mimosine-degrading enzyme activity in some Rhizobium strains. Appl Environ Microbiol 60:4268–4272
     [Google Scholar]
  31. Stachel S.E., An G., Flores C., Nester E.W. 1985; A Tn 3lacZ transposon for the random generation of β -galactosidase gene fusions: application t o the analysis of gene expression in Agrobacterium . EMBO J 4:891–898
     [Google Scholar]
  32. Vincent J.M. 1970; A Manual for the Practical Study of Root- Nodule Bacteria (IBP Handbook no. 15). Oxford:. Blackwell Scientific Publications.164
     [Google Scholar]
  33. Zhan H., Gray J.X., Levery S.B., Rolfe B.G., Leigh J.A. 1990; Functional and evolutionary relatedness of genes for exopolysaccharide synthesis in Rhizobium meliloti and Rhizobium sp. strain NGR234. J Bacteriol 172:5245–5253
     [Google Scholar]
  34. Zevenhuizen L. P. T. M., van Neerve A. R. W. 1983; (1→2) β - D-glucan and acidic oligosaccharides produced by Rhizobium meliloti .. Carbohydr Res 118:127–134
     [Google Scholar]
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The symbiotic phenotypes of exopolysaccharide-defective mutants of Rhizobium sp. strain TAL1145 do not differ on determinate- and indeterminate-nodulating tree legumes
Microbiology 143, 1959 (1997); https://doi.org/10.1099/00221287-143-6-1959
/content/journal/micro/10.1099/00221287-143-6-1959
/content/journal/micro/10.1099/00221287-143-6-1959
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