Article Figures & Data
Figures
- Figure 1.
Typical HPLC-PDA chromatograms of aqueous 80% methanol extracts of leaves of rose at 360 nm for Si− uninoculated (A), Si+ uninoculated (B), Si− inoculated (C), and Si+ inoculated (D). Compounds are as follows: 1, unknown phenolic acid; 2, 3-O-caffeoylquinic acid (neochlorogenic acid); 3, 5-O-caffeoylquinic acid (chlorogenic acid); 4, quercetin-3-O-gentiobioside; 5, quercetin diglycoside; 6, quercetin derivative; 7, quercetin pentoside; 8, quercetin-3-O-galactoside (hyperoside); 9, quercetin-3-O-rutinoside (rutin); 10, quercetin-3-O-glucoside (isoquercitrin); 11, quercetin-3-O-arabinoside (avicularin); 12, quercetin-3-O-rhamnoside (quercitrin); 13, kaempferol-3-O-pentoside; and 14, kaempferol-3-O-rhamnoside (afzelin). The chromatographic conditions and the validation of the HPLC method are described in “Materials and Methods.” mAU, Milliabsorbance units.
- Figure 2.
Contents of total phenolic acids (A) and 5-O-caffeoylquinic acid (chlorogenic acid; B) in leaf extracts of rose from plants either treated with (Si+) or without (Si−) Si followed by either inoculation with P. pannosa or no inoculation. Data represent results from one experiment, and each observation represents the mean from three extractions. All values are presented as means ± se. Means within each time point are comparable, and bars marked by different letters are significantly different. Further information on the results from this experiment is given in Supplemental Table S1. The findings of this experiment were confirmed in a second independent experiment.
- Figure 3.
Contents of total flavonoids and selected flavonoids in leaf extracts of rose from plants either treated with (Si+) or without (Si−) Si followed by either inoculation with P. pannosa or no inoculation. A, Total flavonoids. B, Quercetin-3-O-galactoside (hyperoside). C, Quercetin-3-O-rutinoside (rutin). D, Quercetin-3-O-arabinoside (avicularin). E, Quercetin-3-O-rhamnoside (quercitrin). F, Kaempferol-3-O-rhamnoside (afzelin). Data represent results from one experiment, and each observation represents the mean from three extractions. All values are presented as means ± se. Means within each time point are comparable, and bars marked by different letters are significantly different. Further information on the results from this experiment is given in Supplemental Table S2. The findings of this experiment were confirmed in a second independent experiment.
- Figure 4.
Powdery mildew severity in leaves of rose after spraying or leaf infiltration with chlorogenic acid and rutin. Control plants were treated with water. Disease severity was scored 9 d after inoculation with P. pannosa. Data represent results from one experiment, and each observation represents the mean from 22 leaves. All values are presented as means ± se. Means within each application method are comparable, and bars marked with different letters are significantly different. The findings of this experiment were confirmed in a second independent experiment.
Tables
- Table I. LC-PDA-MS analysis (UV spectra, characteristic ions, and molecular masses) of phenolic acids and flavonoids in aqueous 80% methanol extracts of leaves of rose
Data represent results from the analysis of samples from two independent experiments, each with three independent extractions. Compounds listed here were detected in all samples.
Peak No.a Rtb LC-MS (Atmospheric Pressure Chemical Ionization, Negative Ion Mode) HPLC-PDA, UV Spectra, λmax Compound min m/z (% base peak) nm 1 10.5 347 [M–H]– (56), 301 (52), 139 (100) 296sh,c 323 Unknown phenolic acid 2 12.4 353 [M–H]– (100), 191 (18), 179 (15) 302sh, 329 3-O-Caffeoylquinic acid (neochlorogenic acid)d 3 18.6 353 [M–H]– (100), 325 (29), 191 (19) 298sh, 325 5-O-Caffeoylquinic acid (chlorogenic acid)d 4 25.6 625 [M–H]– (100), 463 (28), 301 (9) 253, 263sh, 353 Quercetin-3-O-gentiobiosidee 5 31.3 609 [M–H]– (100), 447 (21), 301 (8) 265, 284sh, 348 Quercetin diglycosideef 6 41.8 615 [M–H]– (100), 493 (4), 463 (7), 441 (9), 301 (24) 262, 291sh, 352 Quercetin derivativef 7 42.9 433 [M–H]– (7), 301 (100) 255, 285sh, 348sh, 362 Quercetin pentosidef 8 44.7 463 [M–H]– (100), 301 (6) 253, 299sh, 356sh, 364 Quercetin-3-O-galactoside (hyperoside)d 9 47.2 609 [M–H]– (100), 463 (67), 301 (19) 256, 263sh, 298sh, 354 Quercetin-3-O-rutinoside (rutin)d 10 48.3 463 [M–H]– (100), 301 (22) 256, 263sh, 299sh, 354 Quercetin-3-O-glucoside (isoquercitrin)d 11 54.7 433 [M–H]– (100), 301 (7) 256, 263sh, 301sh, 352 Quercetin-3-O-arabinoside (avicularin)d 12 56.5 447 [M–H]– (100), 301 (10) 256, 262sh, 305sh, 348 Quercetin-3-O-rhamnoside (quercitrin)d 13 63.5 417 [M–H]– (100), 285 (10) 264, 294sh, 331sh, 345 Kaempferol-3-O-pentosidef 14 66.0 431 [M–H]– (100), 285 (9) 263, 296sh, 324sh, 343 Kaempferol-3-O-rhamnoside (afzelin)d ↵a Peak numbers correspond to the compound numbers in Figure 1 and Supplemental Figure S2.
↵b Rt, Retention time on HPLC.
↵c sh, Shoulder.
↵d Conclusively identified by comparison with authentic standard.
↵e Identification based on comparison of retention time, UV, and LC-MS data with data from the literature (Masada et al., 2009).
↵f Tentatively identified by UV and mass spectral data.
- Table II. Quantitative recordings of infection biology of P. pannosa and defense responses in the fifth developed leaves of rose cv Smart
Plants were treated with chlorogenic acid and rutin applied by spraying or leaf infiltration. Control plants were similarly treated with water. Observations were made at 72 hai, and values were calculated on the basis of the number of germinated conidia. Data represent results from one experiment, and each observation represents the mean from three leaves. All values are presented as means ± se. The findings of this experiment were confirmed in a second independent experiment.
Application Treatment Odds Ratioa Chlorogenic Acid Rutin Control Chlorogenic Acid Rutin Control Sprayingb Germinated 15.3 ± 0.33 14.7 ± 0.33 25.3 ± 0.33 0.53*** 0.51*** 1.00 With appressoria 10.7 ± 0.33 10.2 ± 0.58 22.0 ± 0.58 0.43*** 0.40*** 1.00 With haustoria 8.7 ± 0.67 9.2 ± 0.33 13.3 ± 0.33 0.52NS 0.56NS 1.00 With ESH 8.7 ± 0.67 9.2 ± 0.33 13.3 ± 0.33 0.52NS 0.56NS 1.00 With FEC 8.0 ± 0.00 8.1 ± 0.00 7.9 ± 0.67 1.00NS 1.02NS 1.00 With papillae 8.7 ± 0.67 8.6 ± 0.33 9.3 ± 0.33 0.92NS 0.93NS 1.00 Infiltrationc Germinated 15.3 ± 0.33 16.7 ± 0.33 24.7 ± 0.33 0.55*** 0.60*** 1.00 With appressoria 10.7 ± 0.33 11.9 ± 0.58 21.4 ± 0.67 0.44*** 0.50*** 1.00 With haustoria 8.0 ± 0.00 10.0 ± 0.00 11.9 ± 0.33 0.65NS 0.82NS 1.00 With ESH 8.0 ± 0.00 10.0 ± 0.00 11.9 ± 0.33 0.65NS 0.82NS 1.00 With FEC 9.3 ± 0.33 10.7 ± 0.33 9.4 ± 0.33 0.99NS 1.15NS 1.00 With papillae 9.3 ± 0.33 10.7 ± 0.33 10.0 ± 0.00 0.93NS 1.07NS 1.00 ↵a Odds ratio for comparison of treatments (control used as a reference; odds ratio = 1.00). NS, Nonsignificant difference; *** significant at P < 0.001; * significant at P < 0.05.
↵b Sprayed with a solution (1 mg mL−1) of chlorogenic acid or rutin until runoff.
↵c Infiltrated with a solution (1 mg mL−1) of chlorogenic acid or rutin.
- Table III. Quantitative real-time RT-PCR analysis of PAL, CHS, and CAD gene expression in leaves of rose from plants either treated with (Si+) or without (Si−) Si followed by either inoculation with P. pannosa or no inoculation
Values shown represent fold up- or down-regulation in Si− inoculated, Si+ uninoculated, and Si+ inoculated plants relative to Si− uninoculated plants (relative expression ratio = 1) at each time point, after normalization of all treatments to 18S rRNA. Data represent results from one experiment, and each observation represents the mean from three extractions. All values are presented as means ± se. The findings of this experiment were confirmed in a second independent experiment. * Significant change; ns, nonsignificant change.
Genes Si Supply Pathogen Fold Change 0 hai 24 hai 72 hai 120 hai PAL Si− Uninoculated 1.0 1.0 1.0 1.0 Si− Inoculated −1.7 ± 0.17ns 3.7 ± 1.25* 1.5 ± 0.17ns 1.1 ± 0.40ns Si+ Uninoculated −1.6 ± 0.14* 2.6 ± 0.60* 1.2 ± 0.27ns −1.0 ± 0.25ns Si+ Inoculated −1.4 ± 0.11* 5.2 ± 1.49* 39.3 ± 11.6* 3.0 ± 0.58* CHS Si− Uninoculated 1.0 1.0 1.0 1.0 Si− Inoculated 1.3 ± 0.46ns 2.8 ± 1.06* 1.5 ± 0.30* 1.2 ± 0.14ns Si+ Uninoculated −1.4 ± 0.24* 2.6 ± 0.55* 2.6 ± 0.30* 2.2 ± 0.15* Si+ Inoculated −1.0 ± 0.32ns 4.5 ± 0.45* 2.7 ± 0.38* 2.1 ± 0.36* CAD Si− Uninoculated 1.0 1.0 1.0 1.0 Si− Inoculated 1.3 ± 0.21* 1.6 ± 0.59ns −1.8 ± 0.04ns −1.2 ± 0.23* Si+ Uninoculated 2.1 ± 0.40* −0.3 ± 0.26ns −1.1 ± 0.12ns 1.1 ± 0.39ns Si+ Inoculated 1.2 ± 0.28* 2.0 ± 0.17* 2.4 ± 0.55* 2.5 ± 0.40ns
Supplemental Data
Supplemental Figures and Tables
Files in this Data Supplement:
- Supplemental Data - Supplemental Figures 1-3 and Supplemental Tables I and II
