PARP activity and effects on primary metabolism.

<p>The metabolite profile of Arabidopsis shoots was analyzed by GC-MS at the indicated times following transfer of the seedlings after 7 days growth on control media to either control or PARP inhibitor containing media. Plants were grown in parallel to those for the physiological and microarray analysis. Five individual samples, each a pool of 10–12 plants, were harvested and analyzed in three independent experiments (n = 15). Amino, organic and fatty acids with significant changes in their relative abundance are shown. Metabolite content relative to the (−3MB), with red and blue indicating accumulation and depletion, are shown.</p

PARP inhibition induced cell number increase.

<p>Arabidopsis leaf two was analyzed after two days of transferring the seedlings (day2) or after seven days (day7) either to treatment or control conditions. (A) Cell number per mm², (B) cell size in µm and (C) the calculated cell number per leaf is presented. Seedlings were previous grown for seven days in control conditions. In each experiment 4–6 leafs were analyzed and the experiment repeated three times independently (n = 12–18). Significant differences (P<0.05) between treated and untreated plants is indicated by an asterisk.</p

PARP inhibition enhances growth and biomass in control conditions.

<p>Hydroponically grown plants were analyzed in respect to leaf area and biomass. 18 plants per experiment and treatment were analyzed, repeated in four experiments and the average leaf area of plants at day 26 (A) or shoot biomass (B), root biomass (C) and the shoot to root ration (D) of plants harvested day 30 is shown. Significant differences (P<0.05) between treated (0.2 mM 3MB) and untreated (125 µl DMSO) plants is indicated by an asterisk.</p

Ploidy analysis of leaf two.

<p>Arabidopsis leaf two was analyzed at the indicated time points from plants subjected to PARP inhibition or control conditions. (A) the calculated EI is shown, EI represents the number of endo cycles undergone by a typical nuclei (EI  =  1*4N+2*8N+3*16N), in (B–D) percentage of 2N, 4N and 8N nuclei is presented. 10–12 leaf two were pooled for a replicate, four biological replicates were analyzed in each of the two independent experiments (n = 8).</p

PARP inhibition and its effect on the phenylpropanoid pathway.

<p>Arabidopsis shoots were harvested at day7 after transfer. Per replicate 10–12 seedlings were harvested, with 5 biological replicates in each of the three independent experiments (n = 15). The metabolite content relative to the control (no 3MB) is shown and when changed significantly the relative content is underlined. Only metabolites with a clear annotation and positioning within the pathway are shown.</p

PARP inhibition and its effect on guard cell development.

<p>Arabidopsis leaf two was analyzed after two days of transferring the seedlings (day2) or after seven days (day7) either to treatment or control conditions. (A) The stomata index (SI) is shown, calculated by number of stomata divided by number of cells, (B) the number of stomata per mm² and (C) the calculated total number of stomata per leaf is presented. In each experiment 4–6 leafs were analyzed and the experiment repeated three times independently (n = 12–18). Significant differences (P<0.05) between treated and untreated plants is indicated by an asterisk.</p

PARP inhibition affects redox metabolite concentrations.

<p>Arabidopsis shoots were harvested at day7 after transfer. Per replicate 10–12 seedlings were harvested, with 5 biological replicates in each of the three independent experiments (n = 15) and analyzed using LC-MS. The relevant data of the redox metabolites was extracted from the LC-MS data set.</p

MAPMAN metabolic overview of transcriptional changes at day7.

<p>Shown are the overall gene expression changes related to the metabolism day7 between 3MB treated leaves 2 and controls. The Wilcoxon rank sum, with Benjamini-Hochberg correction, is shown below.</p

Altered leaf growth in response to PARP inhibition.

<p>(A–D) Rosettes harvested at the indicated time points were dissected and the single leaf size was measured by ImageJ with (C) indicating cotelydone and (L) indicating leave. 15–25 seedlings were analyzed in an experiment, repeated in four independent experiments (n = 90). Significant differences (P<0.05) between treated and untreated plants is indicated by asterisks. (E) A typical leaf series of 14 day old plants (day7) is shown. The red bar indicates 2 cm.</p

PARP inhibition altering secondary metabolism.

<p>Arabidopsis shoots were harvested at day7 after transfer. Per replicate 10–12 seedlings were harvested, with 5 biological replicates in each of the three independent experiments (n = 15) and analyzed using LC-MS. The five strongest up- and down-regulated metabolites are shown.</p