Article Figures & Data
Figures
- Fig 1
Effects of the HDAC inhibitor TSA on FUM gene expression in wild-type VP2, FR3, and GE1 strains of F. verticillioides. The strains were cultivated in the non-FB-inducing Czapek medium in the absence or in the presence of 1 μM TSA. After 7 days of growth, the amounts of transcripts of FUM1 (A), FUM21 (B), and FUM8 (C) were estimated by the absolute quantification method in RT-qPCR. For each gene, expression was normalized to TUB2 transcript abundance. Mean and standard error were calculated with data from three biological and three analytical replicates. Asterisks indicate a P value of <0.05.
- Fig 2
HPLC quantification of FB1 production in wild-type VP2, FR3, and GE1 strains of F. verticillioides. Values are the means of three independent culture batches for each strain, grown for 7 days in liquid Czapek medium (40 ml, inoculated with 105 spores). The role of HDACs in FB1 production was tested by adding the HDAC inhibitor TSA to the medium to a final concentration of 1 μM. Means and standard deviations were calculated with data from three biological replicates.
- Fig 3
Relative acetylation levels of the FUM1 (A), FUM21 (B), and FUM8 (C) gene promoters under different conditions. Strain VP2 was cultured for 7 days in Czapek medium, in Czapek medium containing 1 μM TSA, an HDAC inhibitor, or in fructose-enriched, FB-inducing medium. Acetylation levels were analyzed by ChIP with antibodies specific to hyperacetylated histone H4 (Penta). The abundance of target DNA was quantified by the comparative CT method (Applied Biosystems) with TUB2 as the endogenous reference for normalization. Error bars indicate the standard errors of two biological and three technical repetitions. Different letters indicate significant differences (P < 0.01). −IND, non-FB-inducing medium; +IND, FB-inducing medium.
- Fig 4
(A) Fluorescence micrographs of the F. verticillioides transformed strain Fv-PFUM1-1 grown in FB-inducing GYAM (pH 3) for 1 week. Conidia, conidiophores, and phialides are the most intensely labeled (white arrows). The lower panels are the corresponding bright-field images. (B) Roots of etiolated maize seedlings colonized by GFP-expressing hyphae; confocal images were taken under UV light 3 weeks after sowing of Fv-PFUM1-1-infected seeds. Blastospores budding from a growing hyphal tip in a xylem vessel are indicated by a white arrow. Plant cell walls fluoresce in red, mycelium in green (total depth, 235 μm; resolution, 0.36 × 0.36 × 1.5 μm).
- Fig 5
FUM1 and/or GFP transcript abundance in the Fv-PFUM1-1 and -3 transformants and in the Fv-PTOXA∷GFP strain grown for 1 week in Czapek medium (non-FB-inducing) (A) and grown for 20 days in fructose-enriched, FB-inducing medium (B). For each gene, transcript amounts were estimated by the absolute quantification method in RT-qPCR after normalization to TUB2 transcripts. Means and standard errors were calculated with data from three biological and three analytical replicates. Asterisks indicate a P value of <0.01.
Additional Files
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
HPLC quantification of FB1 and FB2 production by the F. verticillioides wild-type isolate VP2 and the Fv-PFUM1-1 transformant strain grown on cracked maize kernels for 2 weeks under high humidity.
PDF file, 64K. - Supplemental file 2
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Map of pCAM-PFUM1::GFP.
TIFF file, 6.7MB. - Supplemental file 3
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HPLC quantification of FB1 production in wild-type VP2, FR3, and GE1 strains of F. verticillioides.
Zipped PDF file, 183K. - Supplemental file 4
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Southern blot analysis for the estimation of transgene copy number in transformant F. verticillioides strains Fv-PFUM1-1 and Fv-PFUM1-3.
Zipped PDF file, 1.2MB. - Supplemental file 5
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Supplemental legends.
Zipped PDF file, 11K.
- Supplemental file 1 -
HPLC quantification of FB1 and FB2 production by the F. verticillioides wild-type isolate VP2 and the Fv-PFUM1-1 transformant strain grown on cracked maize kernels for 2 weeks under high humidity.
