Oligonucleotides used in this work.

<p>Oligonucleotides used in this work.</p

Photoactivation of light-inducible genes in the wild type and mutants.

<p>A. Quantitative RT-PCR experiments were performed to measure the relative accumulation of mRNAs in mycelia grown at 22°C for 48 h in the dark (D) or under continuous light (11 W m^-2), or exposed to 30 minutes of light after 48 h of growth in the dark. The plots show the average and standard error of the mean of the relative mRNA accumulation in four independent experiments. The results from each PCR for each gene were normalized to the corresponding PCR for <i>tub-2</i> to correct for sampling errors and normalized to those obtained with the wild type after exposure to 30 min of light. B. A replot of the mRNA accumulation obtained in mycelia grown in the dark using a different scale.</p

RCO-1 and RCM-1 regulate light-dependent gene transcription and photoadaptation.

<p>Quantitative RT-PCR experiments were performed to measure the relative accumulation of mRNA in mycelia of the wild type, a strain with a deletion of <i>rco-1</i> or a strain with the <i>rcm-1</i>^RIP allele grown at 22°C for 48 h in the dark and exposed to white light (1 W/m² blue light) for different times. The plots show the average and standard error of the mean of the relative mRNA accumulation in three independent experiments. The results from each PCR for each gene were normalized to the corresponding PCR for <i>tub-2</i> to correct for sampling errors and normalized to those obtained with the wild type after exposure to 30 min of light.</p

The absence of RCO-1 modifies the kinetics of WCC binding to the promoters of light-regulated genes.

<p>A. WCC binding sites in the promoters of several light-regulated genes. The position of the WCC binding sites in the promoters of <i>frq</i> (proximal site, <i>frq-p</i>; distal site, <i>frq-d</i>), <i>wc-1</i>, <i>vvd</i>, <i>al-1</i> and <i>al-3</i> are shown relative to the initiator ATG. B. Kinetics of WCC binding to the promoters measured by chromatin immunoprecipitation with an antibody against WC-2. Mycelia were grown for 48 hours at 30°C in the dark and then exposed for different times to light. Nuclei were extracted prior to each ChIP experiment. Quantitative PCR were performed to measure the relative accumulation of each DNA segment in immunoprecipitated samples and inputs. Each plot shows the average and standard error of the mean of DNA accumulation in three independent experiments. Results from each PCR for each gene were normalized to the corresponding PCR for 28S DNA to correct for sampling errors, and plotted relative to the amount obtained in the dark.</p

The RCO-1/RCM-1 complex is required for the accumulation of VVD after exposure to light.

<p>(A) The RCO-1/RCM-1 complex is required for the light-dependent mRNA accumulation of <i>vvd</i>. Quantitative RT-PCR experiments were performed to measure the relative accumulation of mRNA in mycelia of the wild type, a strain with a deletion of <i>rco-1</i> or a strain with the <i>rcm-1^RIP</i> allele exposed to white light (1 W/m² blue light) or kept in the dark. The plots show the average and standard error of the mean of the relative mRNA accumulation in three independent experiments. The results from each PCR for each gene were normalized to the corresponding PCR for <i>tub-2</i> to correct for sampling errors and normalized to those obtained with the wild type after exposure to 30 min of light. (B) A reduction in the amount of VVD in the Δ<i>rco-1</i> strain. Strains were ground on Vogel's minimal medium for 48 hours at 22°C in the darkness or under white light (11 W/m²). Then, samples grown in the dark were exposed for 30 minutes to light. Total protein extracts isolated from mycelia of the wild-type strain were separated by SDS–PAGE, and hybridized with anti-WC-1, anti-WC-2 and anti-VVD antibodies. 200 µg of proteins were loaded per lane. The bands that correspond to each protein are marked by an arrow, additional bands are due to WC-2 modification or to the non-specific binding of the WC-1 antibody. The experiment was repeated three times and a representative protein hybridization is shown.</p

Distribution of <i>Neurospora</i> species collected in Spain.

<p>Distribution of <i>Neurospora</i> species collected in Spain.</p

Accumulation of carotenoids in species of <i>Neurospora</i> isolated from Spain.

a<p>Average±SEM (µg/g dry mass).</p

Activation by light of the <i>albino</i> genes in <i>Neurospora</i> species.

<p>Quantitative RT-PCR experiments were performed to measure the relative accumulation of <i>al-1</i> or <i>al-2</i> mRNA in mycelia of wild-type strains exposed to white light (2 W/m² blue light) or kept in the dark. The plots show the average and standard error of the mean of the relative mRNA accumulation in four independent experiments, each with three replicates. The results from each PCR for each gene were normalized to the corresponding PCR for <i>tub-2</i> to correct for sampling errors and normalized to the result obtained with mycelia kept in the dark. The amount of carotenoids accumulated by each wild-type strain in cultures exposed to light is shown under each strain name. The initials describe each <i>Neurospora</i> species: Nc <i>Neurospora crassa</i>, Nt <i>Neurospora tetrasperma</i>, and Nd <i>Neurospora discreta</i>.</p

Survival of <i>Neurospora</i> conidia to UV radiation.

<p>Conidia were exposed to UV radiation during different times or kept unirradiated as a control. Cell viability was assayed after plating irradiated and unirradiated conidia, counting the number of colonies after 2–3 days of growth, and comparing the number of colonies obtained with irradiated and unirradiated conidia. The plot shows the average and standard error of the mean of the percentage of survival to UV in three independent experiments. The amount of carotenoids accumulated by mycelia from each wild-type strain in cultures exposed to light is shown over each strain name. The color used for symbols and lines identify each <i>Neurospora</i> species. The latitudes and longitudes of the isolation sites for each strain are the following: C11A (42.74°, −8.62°), PO10A (42.46°, −8.50°), GC4-1A (27.88°, −15.56°), PO9C (42.46°, −8.50°), GC4-4C (27.88°, −15.56°), TF2-6A (28.37°, −16.65°).</p

Distribution of species of <i>Neurospora</i> across sites in Spain.

a<p>Meters.</p>b<p>The identification of these strains has been reported previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033658#pone.0033658-Jacobson2" target="_blank">[9]</a>.</p