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Figure 4 | BMC Microbiology

Figure 4

From: Rhizobium leguminosarum bv. trifolii rosR is required for interaction with clover, biofilm formation and adaptation to the environment

Figure 4

Sensitivity to antibiotics and profiles of membrane and extracellular proteins of R. leguminosarum bv. trifolii rosR mutants. Relative sensitivity of the R. leguminosarum bv. trifolii rosR mutants to antibiotics, determined by measuring the diameter of growth-inhibition zones (A). The values for the Rt24.2 wild type which were used for result normalization were as follows: chloramphenicol 16.9 ± 1.5 mm, erythromycin 24.0 ± 1.5 mm, gentamicin 22.8 ± 1.8 mm, streptomycin 23.5 ± 2.0 mm, tetracycline 45.2 ± 2.2 mm, polymyxin B 5.5 ± 1.0 mm, ampicillin 9.0 ± 1.0 mm, carbenicillin 24.5 ± 2.5 mm, penicillin G 3.5 ± 0.5 mm, bacitracin 14 ± 2.0 mm. Data shown are means of three replicates. (B) Profiles of membrane and extracellular proteins of the Rt24.2 wild type and Rt2472 rosR mutant grown in TY medium. The migration positions of molecular mass markers are shown. Lanes: 1, 2, 3 - Rt2472 membrane protein fraction: 3 μg, 6 μg, and 9 μg, respectively. Lanes: 4, 5, 6 - Rt24.2 wild type membrane protein fraction: 3 μg, 6 μg, and 9 μg, respectively. Lanes: 7, 8 - Rt2472 extracellular protein fraction isolated from 10 ml and 15 ml culture supernatants, respectively. Lanes: 9, 10 - Rt24.2 extracellular protein fraction isolated from 10 ml and 15 culture supernatants, respectively. The symbols indicate prominent proteins which vary apparently in the amount between the rosR mutant and the wild type: white triangles - proteins up-regulated in Rt2472 mutant, black triangles - proteins of increased amounts in Rt24.2 wild type, arrow - a protein unique to Rt2472 extracellular protein fraction. (C) Membrane and extracellular protein profiles of the wild type and the rosR mutant grown in TY and M1 medium with or without 5 μM exudates. Lane: 1- membrane proteins of Rt2472 grown in TY; 2- membrane proteins of Rt24.2 grown in TY; 3- membrane proteins of Rt24.2 grown in M1; 4 - membrane proteins of Rt24.2 grown in M1 with 5 μM exudates; 5- membrane proteins of Rt2472 grown in M1; 6 - membrane proteins of Rt2472 grown in M1 with 5 μM exudates. In the case of lanes 1 to 6, 5 μg of proteins were used. Lanes 7 and 8 - extracellular proteins isolated from TY supernatant of Rt2472 and Rt24.2 cultures, respectively; Lanes 9 and 10 - Rt24.2 extracellular proteins isolated from M1 and M1 with 5 μM exudates supernatants, respectively; Lanes 11 and 12 - Rt2472 extracellular proteins isolated from M1 and M1 with 5 μM exudates supernatants, respectively. In the case of lines 7 to 12, proteins from 10 ml culture supernatant were used. The asterisks indicate prominent proteins which vary apparently in the amount between TY and M1 media for the wild type and the rosR mutant: red asterisks - proteins unique to Rt24.2 and Rt2472 strains growing in TY medium, yellow asterisk - a protein unique to the extracellular protein fraction of Rt24.2 isolated from TY supernatant, green asterisk - a protein uniquely present in extracellular protein fractions of Rt24.2 and Rt2472 isolated from M1 supernatants, black asterisks - proteins present exclusively in the extracellular protein fraction of Rt24.2 isolated from M1 supernatant.

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