<i>axr6-3</i> and <i>cul1</i><i>-</i><i>7</i> mutations reduce the AR numbers produced by double mutants with <i>sur2</i>.

<p>The number of adventitious roots was counted on at least 35 seedlings of each line in two replicates and the data were pooled. Error bars indicate standard errors. One-way ANOVA and Tukey’s multiple-comparison post-tests indicate that the double mutants are not significantly different from their respective wild types (P<0.05; <i>n></i>70).</p

Identification of EMS-induced mutants by NGS-assisted mapping.

<p>For mutants identified in the reference Col-0 background, taking the green path leads to mutation identification. For mutants identified in a non-reference background, a parallel sequencing of the parental genotype (black path) is required. *Replacement of Col-0 specific SNPs with sites specific for the parental genotype, followed by use of the new constructed genome to extract the EMS-induced mutations in the mutant.</p

Annotation of putative causal mutations (A).

<p>Locations of EMS-induced mutations affecting the CDŚs on chromosome IV are marked with asterisks. The red asterisk indicate the mutation located in the region defined by mapping, which is flanked by the UPSC mapping markers <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0100846#pone.0100846-Pacurar1" target="_blank">[5]</a>. Below, the structure of the <i>CUL1</i> gene is shown, indicating positions of known recessive mutations giving viable mutants used in this study for complementation tests. The position and nature of the <i>cul1-494</i> mutation, compared to wild type are highlighted on the DNA sequence. PCR amplification of the <i>494</i> and <i>sur2-1gl1</i> cDNAs with primers spanning the splicing site affected by the <i>cul1-494</i> mutation (B). A 463 bp fragment, corresponding to the correct spliced variant was detected in <i>sur2-1gl1</i>, while an additional 551 bp splicing variant was detected in <i>494</i>.</p