Validation of the <i>AT2S2</i> promoter sequence.

<p>(A-C) Analysis of relative mRNA accumulation of <i>AT2S2</i> was performed in different plant organs (A), in developing seeds (B), and in developmental series of endosperm (Endo.) and embryo fractions (C). The results obtained were standardized to the <i>EF1αA4</i> (<i>EF</i>) gene expression level. Values are the means and SE of three to six replicates carried out on cDNA dilutions obtained from three independent mRNA extractions. DAA, days after anthesis; Fl, flowers; Rl, rosette leaves; Ro, roots; St, stems. (D-J) Pattern of activity of the <i>ProAT2S2</i>:<i>uidA</i> cassette in maturing embryos harvested 10 (D), 12 (E), 14 (F), or 16 days after anthesis (DAA) (G) and in endosperm fractions harvested 14 (H) or 16 DAA (I). A close-up of a peeled endosperm layer aged 14 DAA is presented in (J). For histochemical detection of GUS activity, tissues were incubated 4 hours in a buffer containing 2 mM each of potassium ferrocyanide and potassium ferricyanide. Microscopy observations were performed using Nomarski optics. Bars = 100 μm in (D-I), 10 μm in (J).</p

Increased omega-7 fatty acid content of <i>ProAT2S2</i>:<i>MYB115</i> seeds.

<p>The five independent transformants considered, TCR4, TAR3, TXR2, TGR5, and TJR2 (in order from left to right), are displayed in the same order in the four graphs in the figure. (A) RT-qPCR analysis of transcript abundance in cDNA prepared from maturing seeds aged 14 DAA to assess efficient overexpression of the transgene. Values are the means and SE of three to 12 replicates performed on cDNA dilutions obtained from three independent mRNA extractions. (B-D) Relative proportion of ω-7 fatty acids (<i>cis</i>-ω-7 C16:1, <i>cis</i>-ω-7 C18:1, and <i>cis</i>-ω-7 C20:1) in mature dry seeds (B), in embryos (C), and in endosperm fractions (D) dissected from mature seeds. Values are the means and SE of five replicates performed on batches of 20 seeds from five plants. Asterisks indicate significant differences from the wild type according to <i>t</i>-test at *** P<0.001 and **P<0.01, respectively.</p

Analysis of <i>MYB115</i>, <i>AAD2</i>, and <i>AAD3</i> transcript levels by quantitative RT-PCR in <i>ProAT2S2</i>:<i>MYB115</i> embryos.

<p>The four independent transformants considered, TAR3, TXR2, TGR5, and TJR2 (in order from left to right), are displayed in the same order in the three graphs in the figure. RT-qPCR analysis of transcript abundance in cDNA prepared from excised embryos aged 14 DAA was carried out to assess efficient overexpression of the transgene (<i>MYB115</i>) and of its targets (<i>AAD2</i> and <i>AAD3</i>). Values are the means and SE of three replicates performed on cDNA dilutions obtained from three independent mRNA extractions. Asterisks indicate significant differences from the wild type according to <i>t</i>-test at *** P<0.001, **P<0.01, and *P<0.05, respectively.</p

Comparison of the omega-7 fatty acid content of seeds from engineered lines of <i>A</i>. <i>thaliana</i> obtained by different groups.

<p>Comparison of the omega-7 fatty acid content of seeds from engineered lines of <i>A</i>. <i>thaliana</i> obtained by different groups.</p

Characterization of transgenic seeds overexpressing <i>MYB115</i>.

<p>The five independent transformants considered, TCR4, TAR3, TXR2, TGR5, and TJR2 (in order from left to right), are displayed in the same order in every graph in the figure. (A) Mature seed dry weight. Values are the means and SE of five replicates carried out on batches of 20 seeds from five plants. (B) Mature seed length. Values are the means and SE of 100 measurements carried out on seeds from five plants. (C) Mature seed width. Values are the means and SE of 100 measurements carried out on seeds from five plants. (D, E) Total fatty acid content of mature dry seeds, expressed in μg.mg^-1 DW (D) or in μg per seed (E). Values are the means and SE of five replicates carried out on batches of 20 seeds from five plants. (F) Total seed fatty acid content of embryos dissected from mature dry seeds. Values are the means and SE of five replicates carried out on batches of 20 embryos from five plants. (G) Total seed fatty acid content of endosperm fractions dissected from mature dry seeds. Values are the means and SE of five replicates carried out on batches of 20 endosperm fractions from five plants. Asterisks indicate significant differences from the wild type according to <i>t</i>-test at *** P<0.001, **P<0.01, and *P<0.05, respectively.</p