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Genetics and Molecular Biology

Phase Variation in Xenorhabdus nematophilus

Antonia Volgyi, Andras Fodor, Attila Szentirmai, Steven Forst
Antonia Volgyi
Department of Biological Sciences, University of Wisconsin, Milwaukee, Wisconsin 53201,and
Department of Genetics, Eotvos Lorand University, Budapest, and
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Andras Fodor
Department of Genetics, Eotvos Lorand University, Budapest, and
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Attila Szentirmai
Department of Microbiology, Kossuth Lajos University, Debrecen, Hungary
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Steven Forst
Department of Biological Sciences, University of Wisconsin, Milwaukee, Wisconsin 53201,and
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DOI: 10.1128/AEM.64.4.1188-1193.1998
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  • Fig. 1.
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    Fig. 1.

    Production of crystal proteins in X. nematophilus. Crystal proteins were isolated and separated by SDS-PAGE as described in Materials and Methods. Bacteria were grown to either the mid-logarithmic phase (lanes 1, 3, 5, 7, 9, 11, 13, and 15) or the stationary phase (lanes 2, 4, 6, 8, 10, 12, 14, and 16). The inclusion body proteins derived from 109 phase I cells are shown in lanes 1, 2, 5, 6, 9, 10, 13, and 14. The proteins derived from 109 phase II cells are shown in lanes 3, 4, 7, 8, 11, and 12. The crystal proteins obtained from N2-4/Iv are shown in lanes 15 and 16.

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    Fig. 2.

    Motility on 0.5% agar. Agar plates were dried, and 1 μl of each bacterial culture was placed on the agar surface. (A) Phase I cells. Colony 1, AN6; colony 2, F1; colony 3, 19061. (B) Phase II cells. Colony 1, AN6; colony 2, F1; colony 3, 19061. (C) Strain N2-4. Colony 1, AN6/II; colony 2, N2-4/Iv; colony 3, N2-4/I.

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    Fig. 3.

    Electron micrographs of AN6/I (A), AN6/II (B), and AN6/II grown in the absence of added NaCl (C). (A) Bar = 2 μm. (B and C) Bar = 1 μm.

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    Fig. 4.

    Outer membrane proteins separated on a urea-SDS-polyacrylamide gel. The proteins produced during the mid-logarithmic phase are shown in lanes 3, 5, 7, 9, 11, and 13. The proteins produced during the stationary phase are shown in lanes 1, 2, 4, 6, 8, 12, and 14. The phase I cell products are shown in lanes 1, 3, 4, 7, 8, 11, and 12. The outer membrane proteins derived from phase II cells are shown in lanes 2, 5, 6, 8, 9, and the outer membrane proteins derived from N2-4/Iv are shown in lanes 13 and 14. The various outer membrane proteins are represented by A, B, S, T, N, and P.

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    Fig. 5.

    Cumulative production of infective juvenile nematodes grown on X. nematophilus strains. Data points represent average values from triplicate samples. The experiment was repeated twice with very similar results.

Tables

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  • Table 1.

    Phenotypic characteristics of phase I, phase II, and variant forms ofX. nematophilus strainsa

    StrainPhaseDye bindingCrystal proteinAntibioticsMotilityOpnBVirulenceNematode production
    Soft agarLB mediumLB medium without NaCl
    AN6IB++++++++
    IIR−−−−+−−+
    F1IB++++++++
    IIR−−/+−−−−++
    19061IB++++++++
    IIR−−−++−++
    N2-4IB++++++++
    IvR++++++−+
    • ↵a B, blue; R, red; +, characteristic apparent; −, characteristic not apparent; −/+, characteristic weakly present. The assays used to determine individual characteristics are described in Materials and Methods.

  • Table 2.

    Pathogenicity of X. nematophilus strains: injection of bacteria into the hemocoels of fifth-instar Manduca sexta larvae

    StrainPhaseNo. of bacteria injectedAvg wt of injected Manduca sexta larvae (g)LT50(h)MortalityaTime of death of last larvae (h)
    AN6I201.85296/644
    II101.591/5
    II1001.731/5
    II4001.77853/6
    F1I2502.00304/430
    II2802.09294/430
    19061I2602.15344/446
    II2701.95294/432
    N2-4I2501.87295/634
    Iv3201.351/6
    • ↵a Number of larvae that died/number of larvae injected.

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Phase Variation in Xenorhabdus nematophilus
Antonia Volgyi, Andras Fodor, Attila Szentirmai, Steven Forst
Applied and Environmental Microbiology Apr 1998, 64 (4) 1188-1193; DOI: 10.1128/AEM.64.4.1188-1193.1998

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Phase Variation in Xenorhabdus nematophilus
Antonia Volgyi, Andras Fodor, Attila Szentirmai, Steven Forst
Applied and Environmental Microbiology Apr 1998, 64 (4) 1188-1193; DOI: 10.1128/AEM.64.4.1188-1193.1998
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