GST pull-down assay.

<p>GST and GST-LS2b fusion proteins incubated with cell lysates expressed His-RPS11 in tubes were the put-in samples transformed onto each right side of the membrane. Glutathione beads added to bind GST and GST-LS2b were washed down, proteins eluted from the beads were the pull-down samples transformed onto each left side of the membrane. The presence of RPS11 was detected by immunoblot with anti-HIS antibody. The presence and expression of GST and GST-LS2b was confirmed by immunoblotting with anti-GST antibody.</p

Interaction between bait LS2b protein and full-length prey protein was re-examined using yeast two-hybrid screening by DDO and QDO selection.

<p>Gradient dilutions of transformation mix were spread respectively on DDO/X and QDO/X/A plates which were cultivated for 5 days at 30°C. Yeast colony contained vectors of pGADT7-RPS11 with pGBKT7-LS2b and pGADT7-2bBP19 with pGBKT7- LS2b were showed on QDO/X/A plates. Vector pGADT7-T with pGBKT7-p53 and pGADT7 with pGBKT7-Lamda were used as positive and negative controls respectively.</p

Viral symptoms of LS-CMV infected on systemic leaves at 7 dpi TRV2-RPS11 <i>N</i>. <i>benthamiana</i> plants and TRV2 controls.

<p>Symptom photos were taken at respectively 5 days, 7days, 15days after CMV mechanically infection. Mock was wild <i>N</i>. <i>benthamiana</i> plant 7 dpi TRV2-RPS11 without CMV infection.</p

The gene silencing suppressor activity of CMV2b protein was reduced by the RPS11 knockdown.

<p>(A) Western blot hybridization analysis of gene silencing suppressor activity of CMVLS2b compared to P19 protein on RPS11 knockdown plants. Total protein samples were extracted 3 days post inoculation of gene silencing suppressor. (B) Further GFP silencing suppressor activity test of LS2b protein was performed with comparison of both P19 and homologous Fny2b proteins. Ponceaus staining of rubisco protein was used to monitor equivalence of protein loading. (C) Quantification for relative GFP accumulation of gene silencing suppressor affected on RPS11 knockdown plants and controls. (D) Comparison of each gene silencing suppressing activity on RPS11 knockdown plants and control. Each value is the mean of three replicates and vertical bars are SD. For a given parameter, means with different letters in C are statistically different at P<0.05. <i>Asterisks</i> in D indicate a significant difference between treatments and the control at P<0.01 (**) according to Student–Newman–Keuls test.</p

Principal component analysis (PCA) of <i>P. sepium</i> leaf samples at all time points of drought and re-watering periods.

<p>(A, C) PC1-PC2 and PC1-PC3 variables loading plots; (B, D) PC1-PC2 and PC1-PC3 samples score plots. (PC1-PC3) the first, second and third principal component; (0.4) −0.4 MPa, (1.0) −1.0 MPa, (+Pro) with 10 mM proline in stress solution, (--Pro) with 10 mM proline in recovery solution; (D24, D96) drought for 24 and 96 hours, (R24, R96) re-watering for 24 and 96 hours after 96 h drought.</p

Significant positive correlation between leaf proline concentration and total antioxidant activity.

<p>Significant positive correlation between leaf proline concentration and total antioxidant activity.</p

Correlation coefficients between Pro and ten antioxidant parameters.

<p>Note: (SOD) superoxide dismutase, (CAT) catalase, (APX) ascorbate peroxidase, (GR) glutathione reductase, (POD) peroxidase, (TFAA) total free amino acids, (SP) soluble sugars, (An), anthocyanin, (TF) total flavonoids, (TP) total phenols.</p

Scree plot of variance explained by each factor of the principal component.

<p>(PC1-PC3) the first, second and third principal component.</p

Proline concentration, Δ¹-pyrroline-5-carboxylate synthetase (P5CS) activity, ornithine δ-aminotransferase (OAT) activity and soluble protein (SP) concentration in the new buds germinated during recovery from −1.0 MPa.

<p>Note: Control, normal mature leaves; ¹ new buds without proline application in recovery solution; ² new buds with 10 mM proline in recovery solution; different letters in the same column indicate significant difference at <i>P</i><0.01.</p

Genetic pattern of male sterility of <i>Salvia miltiorrhiza</i> deduced by segregation result in Table 1 through backcross and testcross.

<p>Genetic pattern of male sterility of <i>Salvia miltiorrhiza</i> deduced by segregation result in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050903#pone-0050903-t001" target="_blank">Table 1</a> through backcross and testcross.</p