Modulation of harpin-triggered apoplastic alkalinisation by different auxins.

<p>Cells were treated with 9 μg ml^-1 harpin (hrp, closed circles) as a positive control, harpin combined with 10 μM (open triangles) or 50 μM auxins (IAA, NAA, or 2, 4-D, closed triangles), or ethanol used as solvent control (con, open circles) in <i>V</i>. <i>rupestris</i> (<b>A</b>, <b>C</b>, and <b>E</b>) and cv. ‘Pinot Noir’ (<b>B</b>, <b>D</b>, and <b>F</b>). Representative experiments from five biological replicas are depicted. Harpin and auxin were added at time 0, if measured isolated, for the combinations, auxins were added 1 h prior to harpin.</p

Parameter analysis for thresholds and on the Kendo and Lovebird test materials in the case of using different QP parameters.

<p>(A) Bad Point Ratio curves for different and QP parameter when is fixed as 8. Experiments are performed on Kendo test sequence. (B) Bad Point Ratio curves for different and QP parameter when is fixed as 8. Experiments are performed on Lovebird test sequence. (C) Bad Point Ratio curves for different and QP parameters when is fixed as 1. Experiments are performed on Kendo test sequence. (D) Bad Point Ratio curves for different and QP parameters when is fixed as 1. Experiments are performed on Lovebird test sequence.</p

Heaviside step function.

<p>Heaviside step function.</p

Working model on the antagonistic interaction of signalling triggered by harpin and auxin.

<p>To reduce complexity, only the earliest events are depicted, omitting ROS activation of calcium influx and effects of rac1-signalling on auxin transport. ① harpin activates the NADPH-dependent oxidase RboH leading to the production of superoxide that can spread in the apoplast. ② Superoxide can penetrate through the plasma membrane (probably by aquaporins) and glutathionylate actin in residue Cys374. This will sequester G-actin from being integrated into the growing end of actin filaments. ③ Alternatively, superoxide can be recruited to transduce the effect of auxin (perceived via the auxin-binding protein, ABP) upon the activation of phospholipase D (PLD) through the small G-protein Rac. ④ PLD will generate phosphatidic acids (PA) that can sequester actin capping proteins (cap) to the membrane, such that elongation of actin filaments is enabled. Alternatively, PA can be partitioned to recruit Rac for the activation of the RboH complex. In this case, the capping proteins will not be recruited to the membrane and constrain the elongation of actin filaments leading as secondary consequence to the formation of thick cables through the activity of severing proteins in combination with free G-actin the formation. ⑤ As third alternative, PA can be converted to PIP2, which will recruit actin-depolymerization factor (ADF) to the membrane. Since ADF is sustaining the monomer turnover at the minus end of actin filaments, this recruitment results in a higher stability of fine cortical actin filaments. The molecular targets for the inhibitors diphenyliodonium (DPI), and <i>n</i>-butanol are inserted in red. Hypothetical aspects of the model that have not been addressed experimentally in plant cells, are indicated by blue question marks: The interaction of Harpin with RboH (①) has not been addressed experimentally so far. Also the glutathionylation of actin in consequence of superoxide penetration (②), so far has been shown for animal systems, but not been addressed in plant cells.</p

The results of each step in our scheme to the depth map reconstruction.

<p>(A) Depth map reconstructed by <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055586#pone.0055586.e015" target="_blank">Equation 1</a>. (B) The enlarged part in (A) for comparison. (C) Depth map reconstructed with MV outlier elimination from our scheme. (D) The enlarged part in (C) for comparison. (E) Depth map reconstructed with absent MV processing but without MV outlier elimination from our scheme. (F) The enlarged part in (E) for comparison. (G) Depth map reconstructed with MV outlier elimination and absent MV processing but without near-boundary depth processing from our scheme. (H) The enlarged part in (G) for comparison. (I) Depth map reconstructed with our scheme. (J) The enlarged part in (I) for comparison. (K) The difference map between (G) and (I) after near-boundary depth processing is applied. (L) The enlarged part in (K) for comparison.</p

Objective results of our method.

<p>(a) Reconstructed quality for 9 consequent depth maps of left and right views for Lovebird1. (b) Quality for 9 synthesized correspondent virtual view images.</p

Experiment settings and coding parameters.

<p>Experiment settings and coding parameters.</p

Effect of Latrunculin and auxins on harpin-induced changes of cell viability.

<p>Cells were treated with Latrunculin B (Lat B, 2 μM), with harpin (hrp, 9 μg ml^-1), or with LatB in combination with harpin and auxin (IAA, NAA or 2, 4-D, 50 μM) following subculture weekly, versus ethanol as solvent control (con) in <i>V. rupestris</i> (A) and cv. ‘Pinot Noir’ (B). Data show mean and standard errors from three independent experiments with 500 cells. Significance levels of differences was analyzed using ANOVA with * significant at P = 5%, and ** significant at P = 1%.</p

Effect of auxins on the induction of defence genes by harpin.

<p>Expression of selected defence genes was conducted by semiquantitative RT-PCR in response to harpin (hrp, 9 μg ml^-1), auxins (IAA, NAA or 2, 4-D, 50 μM), or auxins combined with harpin as compared to ethanol as solvent control (con) in <i>V. rupestris</i> (<b>A</b>) and cv. ‘Pinot Noir’ (<b>B</b>). Transcript abundance was analysed 30 min after addition of harpin preceded by incubation for 60 min with the respective auxins. Genes included <i>StSy</i>, stilbene synthase; <i>PAL</i>, phenylalanine ammonia lyase 1; <i>PR</i>5, <i>PR</i>10, pathogenesis-related proteins 5 and 10). Quantification of transcripts were calculated relative to elongation factor 1α (EF1 α) as internal standard. The data represent mean values from three independent experimental series; error bars show standard errors. Expression difference of defence gene as compared to solvent control were analyzed using ANOVA with * significant at <i>P</i> = 5%, and ** significant at <i>P</i> = 1%.</p

Parameter analysis for and .

<p>(a) The Bad Point Ratio with different selection when is fixed as 4. (b) The Bad Point Ratio with different selection when is fixed as 9.</p