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Isolation of adenylate cyclase mutants from Rhizobium meliloti deficient in nodulation

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Abstract

Six Rhizobium meliloti mutants were isolated after Tn5-mediated mutagenesis as resistant to inhibition by a mixture of amino acids (serine, methionine, glycine and leucine). All were defective in adenylate cyclase activity and failed to form nodules in infected roots of Medicago sativa. Furthermore, like other nodulation mutants, they showed altered motility and increased secretion of exopolysaccharides; addition of cAMP to the growth medium abolished some of these phenotypic defects. The possibility that adenylate cyclase participates in the transduction of signals inducing nodulation is discussed.

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References

  • Alper, M.D. & Ames, B.N. 1978 Transport of antibiotics and metabolite analogs by systems under cyclic AMP control: positive selection of Salmonella typhimurium cya and crp mutants. Journal of Bacteriology 133, 149–157.

    Google Scholar 

  • Beringer, J.E., Beynon, J.L., Buchanan-Wollaston, A.V. & Johnston, A.W.B. 1978 Transfer of the drug-resistance transposon Tn5 to Rhizobium. Nature 276, 633–634.

    Google Scholar 

  • Beuve, A., Boesten, B., Crasnier, M., Danchin, A. & O'Gara, F. 1990 Rhizobium meliloti adenylate cyclase is related to eukaryotic adenylate and guanylate cyclases. Journal of Bacteriology 172, 2614–2621.

    Google Scholar 

  • 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 R. phaseoli and is corrected by cloned DNA from Rhizobium or the phytopathogen Xanthomonas. Molecular and General Genetics 203, 320–323.

    Google Scholar 

  • Cangelosi, G.A., Lynn Hung, V., Puvanesarajah, G., Stacey, D.A., Ozga, D.A., Leigh, J.A. & Nester, E.W. 1987 Common loci for Agrobacterium tumefaciens and Rhizobium meliloti exopolysaccharide synthesis and their roles in plant interactions. Journal of Bacteriology 169, 2086–2091.

    Google Scholar 

  • Carricarte, V.C. 1988. Adenilato ciclasa en plantas superiores: caracterización y purificación de esta enzima en Rhizobium meliloti, nodulos radicalares y Medicago sativa. Thesis, Universidad de Buenos Aires.

  • Catanase, A.C., Emerich, D.W. & Zahler, W.L. 1989 Adenylate cyclase and cyclic AMP phosphodiesterase in Bradyrhizobium japonicum bacteroids. Journal of Bacteriology 171, 4531–4536.

    Google Scholar 

  • Daniel, J. & Danchin, A. 1979 Involvement of cyclic AMP and its receptor protein in the sensitivity of Escherichia coli K12 towards serine. Molecular and General Genetics 176, 343–350.

    Google Scholar 

  • Doherty, D., Leigh, J.A., Glazerbrook, J. & Walker, G.C. 1988 Rhizobium meliloti mutants that overproduce the R. meliloti acidic Calcofluor-binding exopolysaccharide. Journal of Bacteriology 170, 4249–4256.

    Google Scholar 

  • Downie, J.A. & Johnston, A.W.B. 1986 Nodulation of legumes by Rhizobium: the recognized root? Cell 47, 153–154.

    Google Scholar 

  • Dylon, T., Ielpi, L., Standfield, S., Koshyap, L., Douglas, C., Yanofsky, M., Nester, E., Helinski, D.R. & Ditta, G. 1986 Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens. Proceedings of the National Academy of Sciences of the United States of America 83, 4403–4407.

    Google Scholar 

  • Firmin, J.L., Wilson, K.E., Rossen, L. & Johnston, A.W.B. 1986 Flavonoid activation of nodulation genes in Rhizobium reversed by other compounds present in plants. Nature 324, 90–92.

    Google Scholar 

  • Flawia, M.M. & Torres, H.N. 1972 Adenylate cyclase activity in Neurospora crassa. Journal of Biological Chemistry 247, 6873–6879.

    Google Scholar 

  • Geremia, R.A., Cavaignac, S., Zorreguieta, A., Toro, N., Olivares, J. & Ugalde, R.A. 1987 A Rhizobium meliloti mutant that forms ineffective pseudonodules in alfalfa produces exopolysaccharide but fails to form β-(1–2)glucan. Journal of Bacteriology 169, 880–884.

    Google Scholar 

  • Gottfert, M., Horvath, B., Kondorosi, E., Putnoky, P., Rodriguez-Quinones & Kondorosi, A. 1986 At least two nodD genes are necessary for efficient nodulation on alfalfa by Rhizobium meliloti. Journal of Molecular Microbiology 128, 789–798.

    Google Scholar 

  • Gyorgypal, A., Iyer, N. & Kondorosi, A. 1988 Three regulatory nodD alleles of diverged flavonoid specificity are involved in host-dependent nodulation by Rhizobium meliloti. Molecular and General Genetics 212, 85–92.

    Google Scholar 

  • Keller, M., Muller, P., Simon, R. & Puhler, A. 1988 Rhizobium meliloti genes for exopolysaccharide synthesis and nodule infection located on megaplasmid are actively transcribed during symbiosis. Molecular Plant-Microbe Interactions 1, 267–274.

    Google Scholar 

  • Kiely, B. & O'Gara, F. 1983. Cyclic 3,5,adenosine monophosphate synthesis in Rhizobium: identification of a cloned sequence from Rhizobium meliloti coding for adenyl cyclase. Molecular and General Genetics 192, 230–239.

    Google Scholar 

  • Leigh, J.A., Reed, J.W., Hanks, J.F., Hirschand, 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 

  • Lim, S.T. & Shanmugan, K.T. 1979 Regulation of hydrogen utilization in Rhizobium japonicum by cyclic AMP. Biochimica et Biophysica Acta 584, 479–492.

    Google Scholar 

  • McGetrick, A.M., Goulding, C.F., Sundaram, S.M. & O'Gara, F. 1985 Catabolite repression and role of cyclic AMP in CO2 fixation and H2 metabolism in Rhizobium spp. Journal of Bacteriology 163, 1282–1284.

    Google Scholar 

  • Miller, J.H. 1972 In Experiments in Molecular Genetics, pp. 431–432. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

    Google Scholar 

  • Muller, P., Hynes, M., Kapp, D., Niehaus, K. & Puhler, A. 1988 The two classes of Rhizobium meliloti infection mutants differ in exopolysaccharide production and in co-inoculation properties with nodulation mutants. Molecular and General Genetics 211, 17–26.

    Google Scholar 

  • O'Regan, M., Kiely, B. & O'Gara, F. 1989 Expression of the adenyl cyclase-encoding gene (cya) of Rhizobium meliloti F43: existence of two cya genes? Gene 83, 243–249.

    Google Scholar 

  • Paau, A.S., Leps, W.T. & Brill, W.J. 1981 Agglutinin from alfalfa necessary for binding and nodulation by Rhizobium meliloti. Science 213, 1513–1524.

    Google Scholar 

  • Peters, N.K., Frost, J.W. & Long, S.R. 1986 A plant flavonone, luteolin, induces expression of Rhizobium meliloti nodulation genes. Science 233, 977–980.

    Google Scholar 

  • Saier, M.H.Jr, Schmidt, M.R. & Leibowitz, M. 1978 Cyclic AMP-dependent synthesis of fimbriae in Salmonella typhimurum: effects of cya and pts mutations. Journal of Bacteriology 145, 489–493.

    Google Scholar 

  • Salomon, Y., Londos, C. & Rodbell, M. 1974 A highly sensitive adenylate cyclase assay. Annals of Biochemistry 58, 541–548.

    Google Scholar 

  • Selvaraj, G. & Iyer, V.N. 1984 Transposon Tn5 specifies streptomycin resistance in Rhizobium spp. Journal of Bacteriology 158, 580–589.

    Google Scholar 

  • Upchurch, R.G. & Elkan, G.H. 1978 The role of ammonia, L-glutamate, and cyclic adenosine 3′,5′-monophosphate in the regulation of ammonia assimilation in Rhizobium japonicum. Biochimica et Biophysica Acta 538, 244–248.

    Google Scholar 

  • Vincent, J.M. 1975 In Manual for the Practical Study of the Root Nodule Bacteria, IBP Handbook 15, pp. 75–76. Oxford: Blackwell Scientific.

    Google Scholar 

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Bianchini, G.M., Carricarte, V.C., Flawia, M.M. et al. Isolation of adenylate cyclase mutants from Rhizobium meliloti deficient in nodulation. World J Microbiol Biotechnol 9, 168–173 (1993). https://doi.org/10.1007/BF00327829

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  • DOI: https://doi.org/10.1007/BF00327829

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