Fig. 4

aRNA silencing of sidA gene via A. tumefaciens-mediated transformation (ATMT) in P. brasiliensis. a Schematic representation of the T-DNA cassette. The cassette is as following: sidA antisense fragment was placed under control of the calcium binding protein (p-cbp1) promoter of H. capsulatum and the terminator (t-catB) of A. fumigatus. The selection marker, was hygromycin B phosphotransferase (HPH), under control of glyceraldehyde 3-phosphate dehydrogenase of A. nidulans (p-gpdA) with the terminator (t-trpC) of A. nidulans.b Relative quantification performed by qRT-PCR to confirm the gene silencing level in clones transformed with PbSidA-aRNA. The transcript level of PbWT transformed with the empty vector (EV) was also quantified and used as control. The actin gene (XP_010761942) was used as the endogenous control. The Student’s t-test was used for statistical comparisons. **** p < 0.0005. c Growth and viability of P. brasiliensis WT, EV and AsSidA strains. Yeasts strains were cultivated in McMM supplemented with 30 μM FeSO₄ at 37 °C for 192 h. Growth curve profiles were determined by optical density, at a wavelength of 660 nm. Viability was accessed by staining of yeast cells with propidium iodide at 1 μg/ml on the last day of the growth curve. The images were obtained using an Axioscope A1 microscope (Carl Zeiss AG, Germany) and photographed at 493/636 nm