22.二酸化窒素の呼吸器に及ぼす影響に関する形態学的研究(第614回千葉医学会例会・第14回肺癌研究施設例会)

<p>The first two columns are the protein IDs of the JGI <i>P</i>. <i>ostreatus</i> genome database.</p

Genetic effect in growth rate.

a<p>Growth rate in mm/day.</p>b<p>This model considers the sum of the two monokaryotic strains.</p

Time course transcriptional profile of <i>lacc</i> genes in GSC.

<p>The transcript levels were expressed as fold changes (FC) compared to the expression on day 0 (A), and in relative quantities (RQ), which was expressed as a percentage (B). Gene expression ratios with standard errors of the mean are shown in Tables S6 to S9 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073282#pone.0073282.s001" target="_blank">File S1</a>.</p

Non-Additive Transcriptional Profiles Underlie Dikaryotic Superiority in <i>Pleurotus ostreatus</i> Laccase Activity

<div><p>Background</p><p>The basidiomycete <i>Pleurotus ostreatus</i> is an efficient producer of laccases, a group of enzymes appreciated for their use in multiple industrial processes. The aim of this study was to reveal the molecular basis of the superiority of laccase production by dikaryotic strains compared to their parental monokaryons.</p><p>Methodology/Principal Findings</p><p>We bred and studied a set of dikaryotic strains starting from a meiotic population of monokaryons. We then completely characterised the laccase allelic composition, the laccase gene expression and activity profiles in the dikaryotic strain N001, in two of its meiotic full-sib monokaryons and in the dikaryon formed from their mating.</p><p>Conclusions/Significance</p><p>Our results suggested that the dikaryotic superiority observed in laccase activity was due to non-additive transcriptional increases in <i>lacc6</i> and <i>lacc10</i> genes. Furthermore, the expression of these genes was divergent in glucose- <i>vs.</i> lignocellulose-supplemented media and was highly correlated to the detected extracellular laccase activity. Moreover, the expression profile of <i>lacc2</i> in the dikaryotic strains was affected by its allelic composition, indicating a putative single locus heterozygous advantage.</p></div

Time course of laccase activity.

<p>The evolution of the secreted laccase activity during GSC and LSC cultures of the dikaryons 61×63, 36×69, 93×69, 67×69, their corresponding monokaryotic parental strains (mk61, mk63, mk36, mk69, mk93 and mk67), and the model dikaryotic strain N001. The values represented the mean of three biological repetitions. Standard deviations are presented in Table S2 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073282#pone.0073282.s001" target="_blank">File S1</a>.</p

Multivariate analysis of expression profiles.

<p>Cluster analysis of genes and samples using a heat map (A), principal component analysis (B) and Kohonen self-organising map (C). Sample loadings of the two principal components are shown in Table S10 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073282#pone.0073282.s001" target="_blank">File S1</a>.</p

Growth rate^a and RBBR decolorization by <i>P. ostreatus</i> strains.

a<p>Growth rate measured in mm/day.</p>b<p>Groups defined using ANOVA and Tukey <i>post hoc</i> test.</p>c<p>nd = no decolorization.</p>d<p> =  not measured.</p>e<p>Signf. = Significance level.</p

PCR primers, amplicon length, and amplification efficiency of the laccase and reference genes.

a,b<p>Transcript identification numbers correspond to the PC15 version 2.0 and PC9 version 1.0 genomes available at <a href="http://www.jgi.doe.gov" target="_blank">www.jgi.doe.gov</a>.</p>c<p>Only present in the PC15 genome.</p

Selection and validation of the reference genes for qPCR analysis.

<p>The gene expression stability ranking was obtained by GeNorm and NormFinder (A), and NormFinder accumulated standard deviations for the increasing number of reference genes (B). The reference genes, their gene ID in the <i>P. ostreatus</i> PC15 v2.0 genome assembly (<a href="http://genome.jgi-psf.org/PleosPC15_2/PleosPC15_2.home.html" target="_blank">http://genome.jgi-psf.org/PleosPC15_2/PleosPC15_2.home.html</a>), and their functional annotation were: <i>phos</i> (49987, purine phosphorylase), <i>lip</i> (1052421, lipase), <i>pep</i> (1092697, peptidase S9), <i>cyc</i> (1035989, cyclin-like F-box), a<i>ctin2</i> (1114037, actin/actin-like), and <i>cyt-c</i> (1113744, cytochrome c).</p

Genetic effect in enzymatic activities.

a<p>This model considers the sum of the two monokaryotic strains.</p>α<p>Means that the enzymatic activity of this dikaryon is significantly different to both of its monokaryotic counterparts (p<0.05).</p>β<p>Means that the enzymatic activity of dikaryon N001 is significantly different to this new dikaryon (p<0.05).</p