Tilled genes and mutation frequency in the mutant collection with 5530 M2 families screened.

<p>Tilled genes and mutation frequency in the mutant collection with 5530 M2 families screened.</p

Phylogenetic analysis of putative COMT proteins.

<p>Phylogeny tree (phylogram) made with OMT proteins from Brachypodium (BdCOMT), rice (OsOMT), maize (ZmOMT) and Arabidopsis (AtOMT). The proteins known to be involved in lignification in ryegrass (LpCOMT), sorghum (SbCOMT), switchgrass (PvCOMT), fescue (FaCOMT) and poplar (PtCOMT) are included in the analysis and shown in red in the phylogram as well as Arabidopsis (AtOMT1) and Maize (ZmCOMT1) proteins. Brachypodium proteins (BdCOMT) are shown in green. Protein sequences are available in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065503#pone.0065503.s001" target="_blank">Information S1</a>. Bootstrap values indicating the level of support for the displayed representation after re-sampling are shown on each node.</p

Structural representation of the amino acid substitution of BdCOMT6 protein in the line <i>Bd4142</i>.

<p>A. 3-D model of wild-type protein highlighting three amino acids important for proper enzymatic activities. B. 3-D model of the protein in <i>Bd4142</i> line in which Glu-326 is substitued by Lys-326. SAH, S-Adenosyl-L-homocysteine is shown to illustrate the proximity of the substitued amino acid in the substrat binding pocket.</p

Frequencies of induced mutations types in tilled gene-coding regions.

a<p>nucleic acid transition is a non-synonymous mutation and induce amino acid change in the translated protein.</p>b<p>nucleic acid transition produces a stop codon and may induce a truncated protein.</p>c<p>nucleic acid transition is located in splicing motif.</p>d<p>nucleic acid transition induces a synonymous mutation and then no change in the translated protein.</p>e<p>other nucleic acid transition than guanine to adenine and cytosine to thymine.</p

Examples of phenotypes detected in the Brachypodium mutant collection.

<p>A. Spikelets of wild-type plants. B–D Aborted or abnormal spikelets in mutants. E. Ligule of wild-type plant. F. Abnormal ligule in mutant. G, H. Mis-shaped leaves in mutants. I. Crumple stem in mutant. J. Coloured stem in mutant. K. adventitious roots in mutant. L. Curved stems in mutant. M. Segregant phenotype for tiller formation in M2 plants compared to WT (right). N. Floppy stems in mutant.</p

Klason lignin (KL) level of extractive-free mature stems of <i>Bd4142, Bd4604</i> and <i>Bd5139</i> lines as compared to control samples.

<p>Control samples are either wild-type (Bd21-3) or azygous plants grown together with the corresponding mutants. The KL level is expressed as weight percentage of the extractive-free sample. Data are means and SD from 4 or 5 plants analyzed per line. Asterisks indicate significant difference compared to the control (ANOVA value at P<0.05).</p

Relative frequency (% molar) of <i>p</i>-hydroxyphenyl H. guaiacyl G. syringyl S and 5-hydroxyguaiacyl 5-OH G monomers released by thioacidolysis of mature and extractive-free stems from control (Ctrl) and mutant <i>Bd4142</i>, <i>Bd4604</i>, <i>Bd5139</i> lines.

<p>Ctrl (azygous line for <i>Bd4142</i> and wild-type line for <i>Bd4604</i> and <i>Bd5139</i>) and corresponding mutant samples were recovered from plants grown together and in identical conditions.</p>a<p>number of replicates. The data represent the means and SD (between brackets). Asterisks indicate significant differences compared to the corresponding Ctrl (ANOVA, value at P<0.01).</p