Supplemental data for Satterlee et al, 2019
Creators: name: Satterlee, Tim
name: Entwistle, Sarah
name: Yanbin Yin
name: Cary, Jeffrey W.
name: Lebar, Matthew
name: Losada, Liliana
name: Calvo, Ana M.
Publisher: name: figshare
Year: 2019
Subjects: subject: Genetics
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Microbiology

description: Supplemental data for article titled "<i>rmtA</i>-Dependent Transcriptome and its Role in Secondary Metabolism, Environmental Stress, and Virulence in <i>Aspergillus flavus".</i><br>Figure S1- GO Term Enrichment of Differentially Expressed Genes. GO terms associated with DEGs in DEL/WT (left) and in OE/WT (right) comparisons. The count of DEGs in each term is proportional to the heights of the bars. GO terms associated with biological processes are in green, cellular components are in blue, and molecular functions are in red. Down regulated genes are to the left of the origin, up regulated to the right. The top 10 terms for each broad category are presented in the figure for each condition. <br>Figure S2 – Construction of <i>gliP</i> mutant strainRepresentation of the strategy used to create an <i>gliP</i> knockout strain via homologous recombination, replacing <i>gliP</i> with the pyrG selection marker. Primers AFLA_gliP_P1-P8 (P1-P8) were used in the creation of the deletion cassette by fusion PCR. Primers P1 and Afum_pyrG_R (R), PCR amplified a 3.1 kb DNA fragment from the genome of the <i>ΔgliP</i> strain, indicating successful gene replacement.<br>Figure S3 – Evaluation of the effect of <i>rmtA</i> on the resistance to osmotic stress in <i>A. flavus </i>To induce osmotic stress, 1 M sucrose, 1.2 M sorbitol, 0.6 M KCl, and 0.7 M NaCl were added to PDA in each case. Wild type (WT), <i>ΔrmtA</i>, complementation (com), and over-expression rmtA (OE) were point-inoculated and grown at 30 °C in the dark. A) Photographs of the cultures after 48 h of incubation. B) Diameter of fungal colonies growing on PDA plus 0.7 M NaCl . <br>Figure S4 – Assessment of the effect of <i>rmtA</i> on the resistance to different temperatures in <i>A. flavu</i>s Wild type (WT), <i>Δrmt</i>A, complementation (com), and over-expression <i>rmtA</i> (OE) were point-inoculated on PDA and grown at 25 °C, 28 °C, 30 °C, 37 °C, 40 °C and 42 °C in the dark. Photographs of the cultures after 48 h of incubation are shown. <br>Table S1- Differentially expressed gene analysis of <i>rmtA</i> transcriptome. Analysis of differentially expressed genes reported as log fold change.<br>Table S2- Listing of transcription factors that are <i>rmtA-</i>dependent. A list of transcription factor genes obtained from the Fungal Transcription Factor Database and compared to the list of DEGs looking for those that are <i>rmtA</i>-dependent. <br>Table S3- Genes related to enviormental stress that are regulated by <i>rmtA. </i>A list of genes related to enviormental stress in fungi was obtained from Miskei et al.(2009) and compared to the list of <i>rmtA </i>DEGs.<br>Table S4- <i>rmtA</i>-dependent genes that are active during virulence of maize. Differentially expressed <i>rmtA </i>genes were compared to the dataset of genes differentially expressed during maize infection, from Dolezal et al. (2013). Genes were categorized in different groups: those virulence-related genes showing upregulation were compared to <i>rmtA</i> DEGs (both up or down regulated), and virulence-related genes that showed downregulation were also compared to <i>rmtA </i>DEGs (both up or down regulated).<br><br>