Generation of <i>F</i>. <i>oxysporum</i> strains overexpressing <i>ctr3</i> and/or <i>fre9</i> in a <i>mac1</i>Δ background.

<p><b>(A)</b> Physical maps of the DNA constructs used for over-expression of <i>ctr3</i> and <i>fre9</i> in <i>mac1</i>Δ. Relative positions of PCR primers are indicated. <b>(B, C)</b> Agarose gel electrophoresis of PCR products obtained using primer pairs Gpda4 and Ctr3-FOXG_07770-R <b>(B)</b>, and Gpda4 and Fre9-3’-Rn <b>(C)</b> using genomic DNA extracted from the indicated nourseothricin-resistant transformants or from <i>mac1</i>Δ as a negative control. M, molecular size markers. <b>(D, E)</b> Real-time RT-qPCR analysis was performed in the indicated strains grown in the absence (-Cu) or presence (+Cu) of 100 μM CuSO<sub>4</sub>. Transcript levels of the indicated genes are expressed relative to those of the wt strain in -Cu conditions. Bars represent standard deviations (<i>n</i> = 3, biological replicates). <i>p</i>-values: ns>0.05, *<0.05, **<0.01 versus <i>mac1</i>Δ, under the same condition, according to two-tailed unpaired Student’s <i>t</i> test.</p> <p>(PDF)</p>