Figure_6.tif (753.22 kB)

Mrr1 regulates MDR1 phenotypes via modulation of atrB expression.

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posted on 2009-12-18, 00:45 authored by Matthias Kretschmer, Michaela Leroch, Andreas Mosbach, Anne-Sophie Walker, Sabine Fillinger, Dennis Mernke, Henk-Jan Schoonbeek, Jean-Marc Pradier, Pierre Leroux, Maarten A. De Waard, Matthias Hahn

(A) Expression of ABC transporter genes in strains with different levels of mrr1 expression. Mean values are shown from two sensitive strains (B05.10, B05.-Hyg3), two MDR1 strains (D06.5-16, D06.7-27), two MDR1 mrr1 k.o. transformants (D06.5-16(Δmrr1)-5, -7), and two B05.Hyg-3 transformants expressing mrr1^V575M (B05.Hyg-3(+mrr1^V575M)-5, -6). (B) Fludioxonil (¹⁴C-labeled) accumulation by wild type strains and strains with mutations in atrB and mrr1, after 10 min (shaded) and 60 min (white bars). Significant differences between the values of the mutants and those of their parent strains are indicated separately for 10 min and 60 min values (n.s.: not significant; **: p<0.01; ***: p<0.001). (C) Drug sensitivities of atrB and mrr1 mutants. For each mutant, mean values of two or three transformants were used to calculate resistance factors relative to sensitive strain B05.10 or (in case of B05.Hyg-3(+mrr1^V575M) to strain B05.Hyg-3. ^#MDR1 strain D06.7-27 showed higher resistance to cyprodinil (standard deviation = 12.0) and lower resistance to tolnaftate, compared to other MDR1 strains. Significantly different resistance values of the transformants relative to their parent strains are indicated (^O Not significant; * p<0.05; ** p<0.01; *** p<0.001). ^§Due to limited solubility of tolnaftate, no accurate values above 25-fold could be determined. (D) Fungicide sensitivity phenotypes on agar plates. 1: B05.10 (sensitive); 2: B05.10(ΔatrB)-4; 3: B05.10(ΔatrB)-5; 4: B05.10(Δmrr1)-8; 5: B05.10(Δmrr1)-18; 6: D06.5-16 (MDR1); 7: D06.5-16(ΔatrB)-1; 8: D06.5-16(ΔatrB)-2; 9: D06.5-16(Δmrr1)-5; 10: D06.5-16(Δmrr1)-7; 11: B05.Hyg-3 (sensitive); 12: B05.Hyg-3(+mrr1^V575M)-5; 13: B05.Hyg-3(+mrr1^V575M)-6; 14: B05.Hyg-3(+mrr1^V575M)-10. Top: HA, 2.5 d.p.i.; middle: HA, 0.03 mg/l fludioxonil, 4 d.p.i.; bottom: GB5 (glucose), 0.01 mg/l cyprodinil, 4 d.p.i..