Ancestral relationship among worldwide PST populations as inferred from the analyses of Approximate Bayesian Computations.

<p>Ancestral relationship among worldwide PST populations as inferred from the analyses of Approximate Bayesian Computations.</p

Discriminant analysis of principal components (DAPC) analysis of worldwide PST populations sampled from different geographical regions.

<p>The Eigen values of the analysis suggest that the first two components explained the maximum genetic structure of the dataset (A). The Bayesian information criteria (BIC) supported six distinct genetic groups (B). Scatter-plot of the worldwide distribution of PST isolates into six genetic groups (C).</p

Expected (He) and observed (Ho) heterozygosity for clone-corrected data based on 20 polymorphic microsatellite loci for PST isolates sampled from diverse geographical regions.

<p>Expected (He) and observed (Ho) heterozygosity for clone-corrected data based on 20 polymorphic microsatellite loci for PST isolates sampled from diverse geographical regions.</p

Estimates of F_ST (upper diagonal) and its significance (lower diagonal) based on 20 microsetillite loci for 386 PST isolates representing worldwide geographically spaced populations.

<p>The lower two lines shows F_ST and its p-value for isolates representing the post-2000 emerged strains. Non-significant <i>F_ST</i> values (>0.01) are shown in bold.</p

Clustering of 409 PST isolates representing worldwide geographical regions to genetic groups for the optimal K-value (K = 6) in the DAPC analysis.

<p>PstS1 and PstS2 refers to the two closely related aggressive strains, while PstS3 refers to the older aggressive isolates regularly reported in Southern Europe.</p

Sampling regions and number of isolates selected for the analysis of the worldwide population structure of wheat yellow rust pathogen <i>Puccinia striiformis</i> f.sp. <i>tritici</i>.

<p>Sampling regions and number of isolates selected for the analysis of the worldwide population structure of wheat yellow rust pathogen <i>Puccinia striiformis</i> f.sp. <i>tritici</i>.</p

<i>Botrytis cinerea</i> strains used in this study.

<p>Fitness characteristics were measured as indicated in Material in Methods.</p

Heterologous expression of <i>bcbik4</i> and <i>bcbik5</i> of the gray strain B05.10 in the respective <i>Fusarium fujikuroi</i> (IMI58289) deletion mutants.

<p>A. Expression of <i>ffbik2</i> and <i>ffbik4/bcbik4.</i> Wild type (IMI58289) and mutants were grown for three days in 10% liquid ICI (0.6 mM glutamine) medium at 28°C and 180 rpm. The northern blots were hybridized with probes for <i>ffbik2</i> or simultaneously hybridized with probes for <i>ffbik4</i> and <i>bcbik4</i>. rRNA is shown as loading control. B. Identification of bikaverin and norbikaverin production. Wild type (IMI58289) and mutants were grown in liquid ICI medium containing 6 mM glutamine for three days. Samples were taken and used for HPLC-DAD analyses (detected at 450 nm) as described in Material and Methods. Peaks for norbikaverin (1) and bikaverin (2) are labeled in the chromatograms.</p

Fungal strains used in this study for genetic modifications.

<p>Fungal strains used in this study for genetic modifications.</p

Role of the global regulator BcVEL1 in the pink strain 1750.

<p>A. Phenotypes caused by the deletion of <i>bcvel1</i> in two different genetic backgrounds, 1750 and B05.10. Virulence of strains was monitored on living plants of <i>Phaseolus vulgaris</i> (7 days after inoculation), sclerotial development by incubation on complete medium (CM) in continuous darkness (14 days), oxalic acid production by using the pH indicator bromothymol blue (color change from green to yellow indicate the acidification of the culture medium) (7 days), bikaverin formation by incubation in liquid or on solid CD-B medium in the dark (7 days), and melanin formation by incubation in liquid CD-M medium in light-dark conditions (7 days). B. Sequencing of <i>bcvel1</i> in the pink strains 1750 and 1787 revealed several SNPs. The point mutation of base pair 367 in 1750 results in a stop codon; a similar mutation leading to a truncated BcVEL1 was recently found in strain T4 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053729#pone.0053729-Schumacher1" target="_blank">[32]</a>. Both 1750 and 1787 comprise additional point mutations that are either silent or result in amino acid exchanges (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053729#pone.0053729.s002" target="_blank">Fig. S2</a>). C. Detection of <i>bcbik1-6 expression in pink (1750, 1787) and gray (B05.10) strains.</i> Strains were grown for 3 days on solid CD-B medium covered with cellophane overlays. Northern blots were hybridized with probes for <i>bcvel1</i>, <i>bcpks13</i> (encoding the polyketide synthase that catalyzes the first step in melanin biosynthesis), <i>bcbik1-6</i>, and <i>bcactA</i> (actin). rRNA is shown as loading control. D. Effect of fenhexamid on growth rates of 1750 and 1750: Δ<i>bcvel1</i> mutants. Strains were grown on complete medium (CM), CD-M (bikaverin-non-inducing conditions) or CD-B (bikaverin-inducing conditions) supplemented with the indicated concentrations of fenhexamid. Diameters of six colonies per strain and condition were measured after 7 days of incubation; pictures were taken after 13 days.</p