Comparison of expected and observed types of mutations in tilled exonic regions.

<p>The percentage of expected mutations was calculated based on the CODDLE analysis of the tilled exonic regions.</p

Protein analysis of <i>Sl</i>-eIF4E1 G1485A splicing mutant.

<p>(<b>A</b>) Western blot analysis of total soluble leaf protein of <i>N. tabacum</i>, <i>S. lycopersicum</i> Hm-WT and <i>Sl</i>-eIF4E1 G1485A mutant probed with an antibody raised against <i>N. tabacum</i> Nt-eIF4E1. (<b>B</b>) Soluble protein extracts of the Hm-WT and G1485A mutant were purified by affinity chromatography on m7G-sepharose column. Total protein extract (lane 1), the flow through (lane 2), the wash (lane 3) and the bound eIF4E proteins eluted with an m7GDP-cap analogue were analysed by Western blot, using Nt-eIF4E antibody.</p

Tilled genes and mutation density in the M82 mutant population.

<p>M82 mutant population was screened for mutations in the listed genes. The total size of the screened amplicons, for each gene, the number of mutants identified and the mutation frequency for each amplicon are indicated. The average mutation frequency was estimated to one mutation per 574 kb and is calculated as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011313#pone.0011313-Greene1" target="_blank">[21]</a>, except that the sizes of all the amplicons were summed and divided by the total number of identified mutants.</p

Mutations discovered in <i>Sl-eIF4E1</i>, <i>Sl-eIF4E2</i>, <i>Sl-eIF(iso)4E</i>, <i>Sl-eIF4G</i>, and <i>Sl-eIF(iso)4G</i>.

<p>Only exonic mutations are shown. The position of the mutations are indicated relative to the first base of the GenBank sequences.</p

Representation of induced mutations in <i>Sl-eIF4E1</i>, <i>Sl-eIF4E2</i>, <i>Sl-eIF(iso)4E</i>, <i>Sl-eIF4G</i> and <i>Sl-eIF(iso)4G</i>.

<p>Black boxes represent the exons. Lanes linking exons indicate introns. Dashed lines indicate the genomic regions screened for mutations. Triangles pointing up indicate mutations in coding regions, whereas those pointing down indicate mutations in noncoding regions. Red, black and grey triangles represent alterations causing truncations, missense and silent mutations, respectively. Only exons 7 to 9 are shown for <i>Sl-eIF4G</i>.</p

Primers used to amplify <i>Sl-eIF4E1</i>, <i>Sl-eIF4E2</i>, <i>Sl-eIF(iso)4E</i>, <i>Sl-eIF4G</i> and <i>Sl-eIF(iso)4G</i> genes and the size of the tilled amplicons.

<p>*F and R indicate forward and reverse primers, respectively.</p

Potyvirus resistance assays of the <i>Sl</i>-eIF4E1 G1485A splicing mutant.

<p><i>Sl</i>-eIF4E1 G1485A mutant and the corresponding Hm-WT were inoculated with PVY-LYE90, PVY-LYE84, TEV-HAT or PepMoV-Texas. (<b>A</b>) At 15 dpi, plants were assayed for potyviral coat protein accumulation by DAS-ELISA in non-inoculated leaves. (<b>B</b>) PVY-LYE90 and PepMoV RNA accumulation was assessed by RT-PCR in inoculated (il) and systemic leaves (sl). Mock indicates non inoculated M82 plants.</p

EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility-4

<p><b>Copyright information:</b></p><p>Taken from "EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility"</p><p>http://www.biomedcentral.com/1471-2229/7/34</p><p>BMC Plant Biology 2007;7():34-34.</p><p>Published online 21 Jun 2007</p><p>PMCID:PMC1914064.</p><p></p>melon leaf showing systemic MNSV-induced symptoms at 14 days after inoculation. () A melon leaf showing systemic CVYV-induced symptoms at 12 days after inoculation

EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility-1

<p><b>Copyright information:</b></p><p>Taken from "EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility"</p><p>http://www.biomedcentral.com/1471-2229/7/34</p><p>BMC Plant Biology 2007;7():34-34.</p><p>Published online 21 Jun 2007</p><p>PMCID:PMC1914064.</p><p></p>). Primers used in EcoTILLING are complementary to non-coding regions of the gene and are indicated by arrows. Amplified regions are represented by black lines. Sizes (bp) of PCR products are indicated below the lines. Sizes (bp) of exons and introns are also indicated

EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility-3

<p><b>Copyright information:</b></p><p>Taken from "EcoTILLING for the identification of allelic variants of melon , a factor that controls virus susceptibility"</p><p>http://www.biomedcentral.com/1471-2229/7/34</p><p>BMC Plant Biology 2007;7():34-34.</p><p>Published online 21 Jun 2007</p><p>PMCID:PMC1914064.</p><p></p>ned into the binary vector pBIN61 between left (LB) and right (RB) borders of the Ti plasmid. The 35S promoter and terminator are indicated as 35S-P and 35S-T, respectively. () RT-PCR detection of MNSV accumulation in bombarded leaves. pBMα5 (Mα5) and pB264 (264) constructs were bombarded separately and in combination with pB4E-PI (4E-PI) or pB4E-Ved (4E-Ved) into leaves of . Two to three independent samples were included in the gel showed. Virus accumulation was assessed using RT-PCR two days post bombardment. C+ and C- indicate positive and negative controls of RT-PCR, respectively. C+ corresponds to leaves from susceptible melon bombarded with pBMα5 and pB264. In C-, RT-PCR was carried out with RNA from non-inoculated leaves