MYB30 increases 1.0 kb <i>FT</i> promoter activity.

<p>Leaves from Col plants grown in SDs were bombarded with particles carrying <i>1.0kbFT_prom::GreenLUC</i> combined with <i>35S_prom::MYB30,35S_prom::CO</i> or both as indicated. <i>35S_prom::RedLUC</i> was included to measure transformation efficiencies. Values are shown as relative GreenLUC compared to RedLUC signals (top panel) after a 16 h–24 h incubation in constant light conditions. Fold-induction over the baseline level obtained for <i>1.0kbFT_prom::GreenLUC</i> alone is indicated in the table below the graph. Values are averages of measurements of five independent leaves from two technical replicates of one bombardment experiment. The experiment was repeated three times with similar results.</p

<i>SUC2_prom::MYB30</i> expression accelerates flowering dependent on <i>FT</i> and independent on <i>CO</i>.

<p>Flowering time measurement of Col, <i>SUC2_prom::MYB30</i>, <i>ft</i>, <i>SUC2_prom::MYB30;ft</i>, <i>co</i> and <i>SUC2_prom::MYB30;co</i> under LDs (<b>a</b>) and SDs (<b>b</b>) in the greenhouse. Plants <i>SUC::MYB30</i>;<i>ft</i> and <i>SUC2_prom::MYB30</i>;<i>ft;tsf</i> were grown in LDs greenhouse (<b>c</b>). Statistical significance was determined using one way Analysis of Variance (ANOVA) followed by multiple comparison of Holm-Sidak method (p<0.01). Significant differences are indicated by different letters above the bars. The number of plants for each genotype (n) is indicated below the graph.</p

Expression of <i>FT</i> and <i>TSF</i> in WT, <i>ft</i> and <i>co</i> plants.

<p>Material from 13-day-seedlings grown on soil in LDs in the greenhouse was collected at ZT16. <i>FT</i> (<b>a</b>) and <i>TSF</i> mRNA (<b>b</b>) levels were determined by RT-qPCR in different genotypes as indicated. Insert in (<b>a</b>) shows values for <i>co</i> and <i>SUC2_prom::MYB30;co</i> at a lower scale. Fold-change <i>FT</i> expression in the <i>SUC2_prom::</i>MYB30 lines compared to the respective control is indicated above the bar. Values are shown as Mean ± SD after normalization of expression with values obtained for <i>PP2A</i>. Experiments were repeated twice with similar results.</p

Flowering time of <i>SUC2_prom::MYB30</i> and <i>myb30</i> mutant plants in LD and SD conditions.

<p>(<b>a</b>)WT and <i>SUC2_prom::MYB30</i> expressing Col plants were grown in LDs (16 h light) and SDs (8 h light) in a temperature controlled climate chamber. The number of rosette and cauline leaves was counted to determine flowering time. Pictures were taken after the WT plants had started to bolt in each condition. Statistical significance was determined using the Student’s t-test (p<0.01). Significant differences are indicated by different letters above the bars, SD and LD treatments are tested as separate experiments. (<b>b</b>) Position of T-DNA insertion lines of <i>myb30</i>. A 400 bp PCR fragment used to detect expression is indicated. (<b>c</b>) Semi-quantitative RT-PCR confirms <i>myb30</i> mutants as loss-of function alleles. (<b>d</b>) Flowering time of Col, <i>myb30_1</i>, <i>myb30_2</i> and <i>SUC2_prom::MYB30</i> plants were measured in LD (left) and SD (right) conditions in a temperature controlled climate chamber. Statistical significance was determined using the Student’s t-test by comparing each genotype to the respective Col control (p<0.01). Significant differences are indicated by different letters above the bars. The number of plants for each genotype (n) is indicated below the graph.</p

<i>SUC2_prom::MYB30</i> promotes flowering independently of SA and FLC.

<p>(<b>a</b>) Flowering time measurement of <i>Col</i>, <i>NahG</i>, <i>SUC2_prom::MYB30</i> and <i>NahG;SUC2_prom::MYB30</i> under LD conditions. (<b>b</b>) Flowering time measurement of Col, <i>SUC2_prom::MYB30</i>, <i>flc</i> and <i>SUC2_prom::MYB30;flc</i> under LD conditions<b>.</b> Statistical significance was determent using one way Analysis of Variance (ANOVA) followed by multiple comparison with the Holm-Sidak procedure (p<0.01). Significant differences are indicated by different letters above the bars. The number of plants for each genotype (n) is indicated below the graph.</p

Expression of flowering time genes in <i>SUC2</i><i>_prom</i><i>::MYB30</i> expressing plants during development.

<p>Material was collected once per week at ZT16 from WT and <i>SUC2_prom::MYB30</i> expressing Col plants grown in LDs in climate chamber. Samples of 1 week and 2 week old plants were collected from all aerial parts, those of 3–5 weeks from leaves. Expression levels were measured by RT-qPCR for <i>FT</i> (<b>a</b>), <i>TSF</i> (<b>b</b>), <i>FLC</i> (<b>c</b>), and <i>SVP</i> (<b>d</b>). Values are shown as Mean ± SD after normalization of expression with values obtained for <i>PP2A</i> for technical triplicates. A biological replicate of the experiment gave similar results.</p

Diurnal expression of <i>MYB30</i>, <i>FT</i> and <i>CO</i>.

<p>(<b>a</b>) Diurnal expression pattern of <i>MYB30</i> was measured in Col grown in LDs and SDs by RT-qPCR. Total RNA was prepared from 10-day old seedlings sampled every 4 hours (ZT0-ZT24). (<b>b</b>) <i>FT</i> expression was measured comparing WT to <i>SUC2_prom::MYB30</i> expressing Col plants. 10-day-plants in LDs were collected every 4 hours from ZT0-ZT24. (<b>c</b>) <i>CO</i> expression was measured comparing WT to <i>SUC2_prom::MYB30</i> expressing Col plants. 10-day-plants in LDs were collected every 4 hours from ZT0-ZT24. Error bars represent the standard error of three technical replicates relative to the expression of <i>PHOSPHATASE 2A</i> (<i>PP2A</i>). The experiment was repeated three times with similar results.</p

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A simple imaging protocol for autofluorescence elimination and optical sectioning in fluorescence endomicroscopy: supplementary materia

<i>A</i>. <i>thaliana PTPLA</i> complements inducible yeast <i>phs1</i> mutant.

<p>(A) PTPLA expression restores growth of <i>Tet-PHS1</i> in presence of DOX. <i>Tet-PHS1</i> was transformed with yeast expression vector pFL61 alone or with yeast <i>PHS1</i>, Arabidopsis <i>PAS2</i> or <i>PTPLA</i>. R1158 is the wild type control strain. (B) Growth kinetic of <i>Tet-PHS1</i> strain expressing PHS1, PAS2 and PTPLA in presence of <i>DOX</i>. Three independent PTPLA expressing clones were analyzed and the mean (+/- sd) is shown. (C-D) Fatty acid content of PTPLA expressing yeasts. PTPLA expression induces fatty acid elongation in yeast <i>Tet-PHS1</i> in presence of <i>DOX</i> (C) and in wild type strain (D). The graph shows FAMES analysis from n = 5–12 and n = 9–15 independent clones for respectively (C) and (D). Data shows means (+/- se). Significant differences between <i>Tet-PHS1</i> (C) or the wild-type (D) and overexpressing strains were determined using the Wilcoxon-test: *p<0,05, **p<0,01, ***p < 0.001.</p

PTPLA is involved in very long chain fatty acids elongation.

<p>(A) 3-hydroxy-acyl-CoA profile of <i>pas2-1</i> and <i>ptpla</i> mutant roots compared to wild type. n = 4. Significant differences were determined using the Wilcoxon-test: *p<0,05, **p<0,01, ***p < 0.001. (B) Three independent experiments showing the VLC/LCFA ratio in <i>pas2-1</i> and <i>ptpla</i> mutant roots compared to wild type. Three independent <i>ptpla</i> mutant lines expressing <i>pPTPLA</i>:<i>PTPLA</i> were used for comparison in the second and third experiments. n = 3. (C) VLC/LCFA ratio in <i>pas2-1</i> and <i>pas2-1/ptpla</i> mutants. n = 3. (D) VLCFA levels in three independent <i>kcr2</i> mutant liness compared to wild type. n = 3. Significant differences were determined using the student’s t-test: *p<0,05, **p<0,01, ***p < 0.001.</p