Figure 1.
Sample pictures of Chrysanthemum × morifolium ‘UBG’ (A) and ‘UPG’ (B) shoot and root systems (C, D) developed from node explants cultured in vitro for 10 weeks on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment; bar = 1 cm. MS, Murashige and Skoog; NPs, nanoparticles; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’; UPG, ‘UTP Pinky Gold’.
Table 1.
Biometric parameters of Chrysanthemum × morifolium ‘UBG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment.
| Treatment | Number of leaves | Micropropagation coefficient | Shoot length (cm) | Shoot FW (mg) | Shoot DW (mg) | Root system FW (mg) | Root system DW (mg) |
|---|---|---|---|---|---|---|---|
| Control | 14.75 ± 0.50 h | 12.00 ± 0.82 i | 5.20 ± 0.70 g | 318.52 ± 128.04 d | 34.10 ± 12.39 e | 14.85 ± 9.12 c | 1.12 ± 0.48 g |
| 100 mg · L−1 ZnO SMPs | 24.25 ± 1.89 b–e | 21.50 ± 1.73 b–f | 9.60 ± 1.70 ab | 916.72 ± 390.89 a–c | 122.80 ± 53.97 a | 94.80 ± 58.17 ab | 7.48 ± 3.97 a–d |
| 200 mg · L−1 ZnO SMPs | 22.50 ± 1.00 b–f | 19.75 ± 0.96 c–h | 9.00 ± 1.93 a–c | 868.42 ± 338.87 a–c | 91.75 ± 44.85 a–e | 94.12 ± 44.37 ab | 7.00 ± 3.13 a–e |
| 400 mg · L−1 ZnO SMPs | 25.25 ± 2.87 a–d | 22.25 ± 2.87 b–e | 10.25 ± 1.95 a | 1,002.95 ± 389.36 a | 120.58 ± 52.61 ab | 107.10 ± 74.91 ab | 8.00 ± 2.98 a–c |
| 100 mg · L−1 ZnO NPs (1.5% H2O) | 22.50 ± 1.73 b–f | 20.00 ± 1.41 c–h | 8.40 ± 1.77 a–d | 531.70 ± 149.45 a–d | 61.52 ± 21.16 b–e | 44.08 ± 27.16 ab | 2.32 ± 1.38 fg |
| 200 mg · L−1 ZnO NPs (1.5% H2O) | 18.50 ± 3.79 gh | 16.50 ± 3.11 h | 6.80 ± 1.35 c–g | 598.95 ± 136.25 a–d | 57.32 ± 12.32 c–e | 90.28 ± 49.57 ab | 4.62 ± 2.33 b–g |
| 400 mg · L−1 ZnO NPs (1.5% H2O) | 19.25 ± 0.96 fg | 16.50 ± 1.29 h | 5.32 ± 0.64 fg | 467.12 ± 222.54 cd | 47.28 ± 17.01 de | 52.85 ± 21.40 ab | 2.98 ± 1.61 e–g |
| 100 mg · L−1 ZnO NPs (6% H2O) | 28.75 ± 2.75 a | 26.00 ± 2.58 a | 8.62 ± 1.68 a–d | 885.00 ± 378.42 a–c | 104.15 ± 61.28 a–d | 54.20 ± 21.37 ab | 4.18 ± 2.82 b–g |
| 200 mg · L−1 ZnO NPs (6% H2O) | 22.00 ± 0.82 c–g | 20.00 ± 0.82 c–h | 6.40 ± 1.75 d–g | 594.45 ± 214.63 a–d | 59.82 ± 22.95 c–e | 66.62 ± 27.71 ab | 3.38 ± 1.23 d–g |
| 400 mg · L−1 ZnO NPs (6% H2O) | 25.75 ± 4.57 a–c | 23.25 ± 4.03 a–c | 7.48 ± 2.09 b–f | 886.85 ± 525.24 a–c | 99.62 ± 56.87 a–d | 133.65 ± 79.22 a | 7.25 ± 3.57 a–e |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 26.00 ± 3.92 ab | 24.00 ± 3.92 ab | 8.98 ± 2.35 a–c | 848.98 ± 418.51 a–c | 107.55 ± 60.71 a–c | 71.10 ± 29.22 ab | 6.35 ± 2.15 a–f |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 24.75 ± 3.30 b–e | 22.50 ± 3.11 a–e | 8.00 ± 1.06 a–d | 699.28 ± 299.83 a–d | 88.62 ± 39.66 a–e | 61.50 ± 35.19 ab | 4.88 ± 2.64 a–g |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 19.50 ± 2.38 fg | 17.00 ± 2.58 gh | 5.25 ± 1.44 e–g | 503.45 ± 253.84 b–d | 60.68 ± 24.29 c–e | 98.58 ± 52.69 ab | 4.70 ± 2.21 b–g |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 21.50 ± 3.70 d–g | 19.50 ± 3.70 d–h | 8.28 ± 2.21 a–d | 822.58 ± 629.57 a–c | 94.30 ± 61.92 a–d | 48.22 ± 24.81 ab | 2.55 ± 1.37 fg |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 22.75 ± 2.22 b–f | 20.25 ± 2.50 c–g | 9.40 ± 1.15 ab | 699.20 ± 213.92 a–d | 73.52 ± 13.02 a–e | 78.00 ± 31.07 ab | 4.52 ± 2.71 b–g |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 22.75 ± 2.36 b–f | 20.25 ± 2.63 c–g | 8.40 ± 1.87 a–d | 798.75 ± 459.66 a–d | 86.70 ± 35.42 a–e | 146.67 ± 131.02 a | 8.32 ± 2.11 ab |
| 100 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 21.25 ± 2.22 e–g | 18.75 ± 1.71 e–h | 9.38 ± 0.95 ab | 849.22 ± 322.33 a–c | 96.32 ± 35.65 a–d | 67.18 ± 33.93 ab | 3.78 ± 1.60 c–g |
| 200 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 21.50 ± 2.08 d–g | 18.75 ± 2.50 e–h | 7.88 ± 0.83 b–d | 643.65 ± 229.90 a–d | 68.72 ± 19.61 a–e | 129.20 ± 53.27 a | 8.15 ± 2.53 ab |
| 400 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 20.00 ± 1.83 fg | 18.00 ± 1.83 f–h | 8.00 ± 1.01 a–d | 788.32 ± 228.25 a–d | 81.00 ± 26.56 a–e | 105.08 ± 42.26 ab | 7.10 ± 3.03 a–e |
| 100 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 19.00 ± 2.94 fg | 16.50 ± 2.65 h | 6.58 ± 0.91 d–g | 528.92 ± 268.54 a–d | 49.15 ± 22.13 c–e | 97.10 ± 40.77 ab | 5.62 ± 1.70 a–f |
| 200 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 21.50 ± 4.36 d–g | 19.00 ± 4.08 d–h | 9.58 ± 2.00 ab | 987.92 ± 370.32 ab | 99.85 ± 38.98 a–d | 143.60 ± 45.49 a | 9.20 ± 3.23 a |
| 400 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 19.50 ± 2.65 fg | 17.25 ± 2.22 gh | 7.60 ± 1.88 b–e | 724.85 ± 535.35 a–d | 71.80 ± 45.90 a–e | 62.32 ± 31.34 ab | 4.33 ± 3.57 b–g |
Table 2.
Biometric parameters of Chrysanthemum × morifolium ‘UPG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment.
| Treatment | Number of leaves | Micropropagation coefficient | Shoot length (cm) | Shoot FW (mg) | Shoot DW (mg) | Root system FW (mg) | Root system DW (mg) |
|---|---|---|---|---|---|---|---|
| Control | 14.75 ± 1.50 e | 11.75 ± 1.50 f | 3.37 ± 0.97 e | 183.88 ± 101.51 c | 30.15 ± 9.71 b | 14.22 ± 3.42 c | 1.82 ± 1.44 d |
| 100 mg · L−1 ZnO SMPs | 18.50 ± 1.00 b–d | 15.50 ± 1.00 c–e | 8.42 ± 1.69 a–d | 621.52 ± 246.24 a–c | 76.82 ± 21.71 ab | 97.58 ± 43.42 b | 6.90 ± 4.00 bd |
| 200 mg · L−1 ZnO SMPs | 17.75 ± 0.96 cd | 14.75 ± 0.96 e | 6.55 ± 2.38 cd | 525.30 ± 281.34 a–c | 59.50 ± 25.42 ab | 130.28 ± 45.46 b | 5.28 ± 2.97 cd |
| 400 mg · L−1 ZnO SMPs | 19.75 ± 1.50 a–d | 16.75 ± 1.50 b–e | 6.85 ± 0.47 cd | 610.65 ± 54.16 a–c | 68.82 ± 9.47 ab | 219.28 ± 82.43 b | 9.15 ± 4.19 a–c |
| 100 mg · L−1 ZnO NPs (1.5% H2O) | 18.75 ± 0.96 b–d | 15.75 ± 0.96 c–e | 6.50 ± 2.43 cd | 479.82 ± 182.32 bc | 58.68 ± 21.99 ab | 102.65 ± 66.78 b | 5.08 ± 1.42 cd |
| 200 mg · L−1 ZnO NPs (1.5% H2O) | 19.00 ± 0.82 b–d | 17.75 ± 2.36 a–c | 7.12 ± 1.66 b–d | 508.12 ± 195.29 a–c | 67.40 ± 25.50 ab | 122.02 ± 50.14 b | 6.72 ± 2.26 cd |
| 400 mg · L−1 ZnO NPs (1.5% H2O) | 20.25 ± 2.50 a–c | 17.25 ± 2.50 a–e | 7.58 ± 2.88 a–d | 665.65 ± 271.19 ab | 82.82 ± 29.18 ab | 91.95 ± 53.37 b | 6.08 ± 3.47 cd |
| 100 mg · L−1 ZnO NPs (6% H2O) | 19.00 ± 1.41 b–d | 16.00 ± 1.41 c–e | 6.30 ± 1.89 de | 543.68 ± 247.53 a–c | 60.38 ± 26.87 ab | 161.10 ± 77.52 b | 7.35 ± 5.83 a–d |
| 200 mg · L−1 ZnO NPs (6% H2O) | 19.75 ± 0.96 a–d | 16.75 ± 0.96 b–e | 7.85 ± 0.79 a–d | 665.58 ± 207.59 ab | 84.38 ± 32.54 ab | 168.35 ± 46.40 b | 7.92 ± 2.66 a–c |
| 400 mg · L−1 ZnO NPs (6% H2O) | 18.50 ± 1.29 b–d | 15.50 ± 1.29 c–e | 6.42 ± 1.50 cd | 511.45 ± 164.74 a–c | 51.02 ± 14.96 ab | 184.92 ± 81.48 b | 6.95 ± 4.82 bd |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 20.50 ± 0.58 ab | 17.75 ± 0.96 a–c | 7.28 ± 1.22 a–d | 753.18 ± 305.43 ab | 74.85 ± 36.24 ab | 101.58 ± 50.87 b | 5.98 ± 2.69 cd |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 18.75 ± 0.50 b–d | 15.75 ± 0.50 c–e | 6.82 ± 1.25 cd | 545.70 ± 131.47 a–c | 61.08 ± 15.74 ab | 229.28 ± 86.24 b | 9.90 ± 4.08 a–c |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 19.75 ± 0.96 a–d | 16.75 ± 0.96 b–e | 7.08 ± 1.28 bd | 678.05 ± 265.25 ab | 77.95 ± 26.21 ab | 186.42 ± 75.81 b | 9.85 ± 4.45 a–c |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 20.50 ± 1.00 ab | 17.50 ± 1.00 a–d | 8.75 ± 1.13 a–d | 755.55 ± 350.31 ab | 85.68 ± 32.51 ab | 140.40 ± 49.83 b | 7.80 ± 3.77 a–c |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 22.25 ± 1.89 a | 19.50 ± 1.73 a | 8.82 ± 1.08 a–c | 858.52 ± 283.60 ab | 99.28 ± 38.17 a | 210.38 ± 64.28 b | 12.98 ± 1.17 a |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 20.75 ± 2.63 ab | 17.75 ± 2.63 a–c | 8.82 ± 1.11 a–c | 746.30 ± 342.36 ab | 83.68 ± 35.37 ab | 482.35 ± 145.86 a | 9.35 ± 4.15 a–c |
| 100 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 19.50 ± 3.11 b–d | 17.25 ± 3.20 a–e | 8.75 ± 1.37 a–d | 618.72 ± 238.85 a–c | 86.40 ± 39.95 ab | 135.98 ± 69.05 b | 8.22 ± 2.35 a–c |
| 200 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 18.50 ± 1.29 b–d | 15.75 ± 0.96 c–e | 8.80 ± 1.70 a–c | 691.82 ± 299.58 ab | 77.88 ± 37.48 ab | 150.38 ± 58.97 b | 8.92 ± 1.29 a–c |
| 400 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 19.00 ± 1.41 b–d | 16.50 ± 1.00 b–e | 9.48 ± 2.30 ab | 764.60 ± 356.38 ab | 78.62 ± 34.05 ab | 137.82 ± 37.44 b | 7.92 ± 1.30 a–c |
| 100 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 19.25 ± 3.77 b–d | 17.25 ± 3.77 a–e | 8.68 ± 3.04 a–d | 687.50 ± 265.24 ab | 81.50 ± 36.31 ab | 104.40 ± 51.14 b | 5.95 ± 5.35 cd |
| 200 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 20.50 ± 1.91 ab | 18.75 ± 1.50 ab | 9.72 ± 1.32 a | 946.78 ± 134.07 a | 406.50 ± 21.20 a | 231.98 ± 133.63 b | 12.62 ± 3.99 ab |
| 400 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 17.25 ± 2.99 de | 15.00 ± 2.58 de | 7.05 ± 2.36 b–d | 733.92 ± 454.71 ab | 74.20 ± 36.72 ab | 146.40 ± 62.51 b | 9.75 ± 4.49 a–c |
Figure 2.
Leaf architecture parameters of Chrysanthemum × morifolium ‘UBG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment. Means ± SD on graphs for each parameter analysed followed by the same letter do not differ significantly at p ≤ 0.05 (Fisher’s test). MS, Murashige and Skoog; NPs, nanoparticles; SD, standard deviation; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’.
Figure 3.
Leaf architecture parameters of Chrysanthemum × morifolium ‘UPG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment. Means ± SD on graphs for each parameter analysed followed by the same letter do not differ significantly at p ≤ 0.05 (Fisher’s test). MS, Murashige and Skoog; NPs, nanoparticles; SD, standard deviation; SMPs, submicron particles; UPG, ‘UTP Pinky Gold’.
Figure 4.
Root system architecture of Chrysanthemum × morifolium ‘UBG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment. Means ± SD on graphs for each parameter analysed followed by the same letter do not differ significantly at p ≤ 0.05 (Fisher’s test). MS, Murashige and Skoog; NPs, nanoparticles; SD, standard deviation; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’.
Figure 5.
Root system architecture of Chrysanthemum × morifolium ‘UPG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment. Means ± SD on graphs for each parameter analysed followed by the same letter do not differ significantly at p ≤ 0.05 (Fisher’s test). MS, Murashige and Skoog; NPs, nanoparticles; SD, standard deviation; SMPs, submicron particles; UPG, ‘UTP Pinky Gold’.
Table 3.
Content and ratios of chlorophylls and carotenoids in Chrysanthemum × morifolium ‘UBG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment.
| Treatment | Chlorophyll a content (mg · g−1 FW) | Chlorophyll b content (mg · g−1 FW) | Chlorophyll a/b ratio | Chlorophyll (a + b) content (mg · g−1 FW) | Carotenoid content (mg · g−1 FW) | Chlorophyll/carotenoid ratio |
|---|---|---|---|---|---|---|
| Control | 1.13 ± 0.16 ab | 0.43 ± 0.07 a–d | 2.63 ± 0.05 bc | 1.56 ± 0.23 ab | 0.25 ± 0.03 ab | 6.24 ± 0.18 c–f |
| 100 mg · L−1 ZnO SMPs | 1.08 ± 0.25 a–c | 0.44 ± 0.09 a–c | 2.45 ± 0.09 e–i | 1.52 ± 0.34 a–c | 0.22 ± 0.06 b–e | 6.91 ± 0.41 b |
| 200 mg · L−1 ZnO SMPs | 0.95 ± 0.15 c–g | 0.39 ± 0.03 c–f | 2.44 ± 0.23 f–i | 1.34 ± 0.18 c–f | 0.21 ± 0.02 c–f | 6.38 ± 0.27 c–f |
| 400 mg · L−1 ZnO SMPs | 0.83 ± 0.08 g–i | 0.30 ± 0.03 j | 2.77 ± 0.08 a | 1.13 ± 0.11 g–i | 0.17 ± 0.02 gh | 6.65 ± 0.25 bc |
| 100 mg · L−1 ZnO NPs (1.5% H2O) | 0.83 ± 0.10 g–i | 0.33 ± 0.04 g–j | 2.51 ± 0.04 d–h | 1.16 ± 0.14 f–i | 0.17 ± 0.02 gh | 6.82 ± 0.26 b |
| 200 mg · L−1 ZnO NPs (1.5% H2O) | 0.63 ± 0.03 j | 0.25 ± 0.02 k | 2.52 ± 0.07 c–h | 0.88 ± 0.05 j | 0.12 ± 0.01 i | 7.33 ± 0.07 a |
| 400 mg · L−1 ZnO NPs (1.5% H2O) | 1.20 ± 0.06 a | 0.46 ± 0.03 a | 2.61 ± 0.05 b–e | 1.66 ± 0.09 a | 0.26 ± 0.02 a | 6.38 ± 0.26 c–f |
| 100 mg · L−1 ZnO NPs (6% H2O) | 0.91 ± 0.06 e–h | 0.35 ± 0.02 e–i | 2.60 ± 0.06 b–f | 1.26 ± 0.07 d–h | 0.18 ± 0.01 f–h | 7.00 ± 0.06 ab |
| 200 mg · L−1 ZnO NPs (6% H2O) | 0.89 ± 0.10 f–i | 0.34 ± 0.03 f–j | 2.62 ± 0.10 b–d | 1.23 ± 0.13 e–i | 0.22 ± 0.05 b–e | 5.60 ± 1.66 f |
| 400 mg · L−1 ZnO NPs (6% H2O) | 0.86 ± 0.07 f–i | 0.32 ± 0.04 h–j | 2.69 ± 0.10 a–c | 1.18 ± 0.11 e–i | 0.18 ± 0.02 f–h | 6.56 ± 0.11 b–d |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 1.04 ± 0.03 b–d | 0.40 ± 0.01 b–e | 2.60 ± 0.10 b–f | 1.44 ± 0.03 b–d | 0.24 ± 0.02 a–c | 6.00 ± 0.34 d–f |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 0.88 ± 0.17 f–i | 0.36 ± 0.05 e–i | 2.44 ± 0.28 f–i | 1.24 ± 0.21 e–h | 0.19 ± 0.03 e–h | 6.53 ± 0.11 b–d |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 0.92 ± 0.11 d–h | 0.38 ± 0.07 d–g | 2.63 ± 0.14 bc | 1.30 ± 0.18 d–g | 0.22 ± 0.03 b–e | 5.91 ± 0.11 ef |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 1.03 ± 0.07 b–e | 0.40 ± 0.04 b–e | 2.58 ± 0.13 b–g | 1.43 ± 0.11 b–d | 0.23 ± 0.01 a–d | 6.22 ± 0.36 d–f |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 0.79 ± 0.12 h–i | 0.31 ± 0.05 ij | 2.55 ± 0.06 b–g | 1.10 ± 0.17 hi | 0.17 ± 0.03 gh | 6.47 ± 0.29 b–e |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 0.98 ± 0.03 c–g | 0.39 ± 0.02 c–f | 2.72 ± 0.04 ab | 1.37 ± 0.05 c–e | 0.22 ± 0.02 b–e | 6.23 ± 0.24 c–f |
| 100 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 0.99 ± 0.06 c–f | 0.38 ± 0.03 d–g | 2.61 ± 0.10 b–e | 1.37 ± 0.09 c–e | 0.22 ± 0.01 b–e | 6.23 ± 0.24 c–f |
| 200 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 1.07 ± 0.06 a–c | 0.45 ± 0.03 ab | 2.38 ± 0.04 hi | 1.52 ± 0.08 a–c | 0.22 ± 0.02 b–e | 6.91 ± 0.20 b |
| 400 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 0.76 ± 0.03 ij | 0.29 ± 0.02 jk | 2.62 ± 0.14 b–d | 1.05 ± 0.04 i | 0.16 ± 0.01 h | 6.56 ± 0.18 b–f |
| 100 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 0.92 ± 0.23 d–h | 0.36 ± 0.08 e–i | 2.56 ± 0.15 b–g | 1.28 ± 0.31 d–h | 0.21 ± 0.06 c–f | 6.10 ± 0.47 d–f |
| 200 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 0.92 ± 0.08 d–h | 0.38 ± 0.02 d–g | 2.42 ± 0.12 g–i | 1.30 ± 0.10 d–g | 0.20 ± 0.02 d–g | 6.50 ± 0.22 b–d |
| 400 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 0.85 ± 0.15 f–i | 0.37 ± 0.08 e–h | 2.30 ± 0.12 i | 1.22 ± 0.22 e–i | 0.20 ± 0.04 d–g | 6.10 ± 0.19 d–f |
Table 4.
Content and ratios of chlorophylls and carotenoids in Chrysanthemum × morifolium ‘UPG’ plantlets developed from node explants cultured in vitro on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment.
| Treatment | Chlorophyll a content (mg · g−1 FW) | Chlorophyll b content (mg · g−1 FW) | Chlorophyll a/b ratio | Chlorophyll (a ± b) content (mg · g−1 FW) | Carotenoid content (mg · g−1 FW) | Chlorophyll/carotenoid ratio |
|---|---|---|---|---|---|---|
| Control | 1.46 ± 0.06 a | 0.51 ± 0.02 a–c | 2.86 ± 0.06 a | 1.97 ± 0.08 a | 0.29 ± 0.01 ab | 6.79 ± 0.07 ef |
| 100 mg · L−1 ZnO SMPs | 1.03 ± 0.04 fg | 0.42 ± 0.03 d–f | 2.45 ± 0.11 d–g | 1.45 ± 0.07 d–f | 0.23 ± 0.01 c–f | 6.30 ± 0.26 ik |
| 200 mg · L−1 ZnO SMPs | 1.08 ± 0.15 e–g | 0.45 ± 0.05 c–f | 2.40 ± 0.06 f–h | 1.53 ± 0.20 c–f | 0.21 ± 0.03 d–g | 7.29 ± 0.48 bc |
| 400 mg · L−1 ZnO SMPs | 1.03 ± 0.16 fg | 0.42 ± 0.08 d–f | 2.45 ± 0.08 d–g | 1.45 ± 0.24 d–f | 0.21 ± 0.04 d–g | 6.91 ± 0.12 d–f |
| 100 mg · L−1 ZnO NPs (1.5% H2O) | 1.16 ± 0.19 c–f | 0.47 ± 0.06 b–e | 2.47 ± 0.09 c–f | 1.63 ± 0.26 b–d | 0.24 ± 0.04 c–e | 6.79 ± 0.30 ef |
| 200 mg · L−1 ZnO NPs (1.5% H2O) | 0.97 ± 0.29 g | 0.40 ± 0.12 ef | 2.42 ± 0.02 e–g | 1.37 ± 0.42 ef | 0.18 ± 0.06 g | 7.61 ± 0.36 a |
| 400 mg · L−1 ZnO NPs (1.5% H2O) | 1.26 ± 0.03 b–d | 0.50 ± 0.03 a–c | 2.52 ± 0.11 b–d | 1.76 ± 0.07 a–c | 0.26 ± 0.01 bc | 6.77 ± 0.15 e–g |
| 100 mg · L−1 ZnO NPs (6% H2O) | 1.40 ± 0.13 ab | 0.53 ± 0.04 ab | 2.64 ± 0.07 b | 1.93 ± 0.17 a | 0.30 ± 0.03 a | 6.43 ± 0.29 hi |
| 200 mg · L−1 ZnO NPs (6% H2O) | 1.14 ± 0.32 c–g | 0.46 ± 0.09 c–e | 2.48 ± 0.23 c–e | 1.60 ± 0.42 bc | 0.25 ± 0.07 c–d | 6.40 ± 0.17 hj |
| 400 mg · L−1 ZnO NPs (6% H2O) | 1.17 ± 0.20 c–f | 0.45 ± 0.07 c–f | 2.60 ± 0.08 bc | 1.62 ± 0.26 b–d | 0.24 ± 0.04 c–e | 6.75 ± 0.06 e–g |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 1.19 ± 0.09 c–f | 0.48 ± 0.03 a–d | 2.48 ± 0.08 c–e | 1.67 ± 0.12 b–d | 0.24 ± 0.01 c–e | 6.96 ± 0.34 de |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 1.09 ± 0.05 d–g | 0.48 ± 0.01 a–d | 2.27 ± 0.06 i | 1.57 ± 0.06 c–f | 0.24 ± 0.02 c–e | 6.54 ± 0.23 e–h |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (1.5% H2O) | 1.16 ± 0.09 c–f | 0.49 ± 0.03 a–c | 2.37 ± 0.08 g–i | 1.65 ± 0.11 b–d | 0.22 ± 0.02 d–g | 7.50 ± 0.19 ab |
| 100 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 1.09 ± 0.18 d–g | 0.46 ± 0.08 c–f | 2.37 ± 0.06 g–i | 1.55 ± 0.26 c–f | 0.25 ± 0.04 c–d | 6.20 ± 0.08 jk |
| 200 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 1.15 ± 0.15 c–f | 0.48 ± 0.05 a–d | 2.40 ± 0.07 f–h | 1.63 ± 0.20 b–d | 0.24 ± 0.03 c–e | 6.79 ± 0.04 ef |
| 400 mg · L−1 ZnO + 0.1% Ag NPs (6% H2O) | 1.30 ± 0.07 a–c | 0.54 ± 0.04 a | 2.41 ± 0.06 e–g | 1.84 ± 0.11 ab | 0.26 ± 0.01 bc | 7.08 ± 0.24 de |
| 100 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 1.02 ± 0.27 fg | 0.42 ± 0.12 d–f | 2.43 ± 0.08 e–g | 1.44 ± 0.39 d–f | 0.20 ± 0.05 e–g | 7.20 ± 0.32 cd |
| 200 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 1.25 ± 0.11 b–e | 0.51 ± 0.03 a–c | 2.45 ± 0.15 d–g | 1.76 ± 0.14 a–c | 0.27 ± 0.02 a–c | 6.52 ± 0.21 f–h |
| 400 mg · L−1 ZnO + 1% Ag NPs (1.5% H2O) | 1.02 ± 0.12 fg | 0.45 ± 0.04 c–f | 2.27 ± 0.09 i | 1.47 ± 0.16 d–f | 0.19 ± 0.03 f–g | 7.74 ± 0.27 a |
| 100 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 1.03 ± 0.07 fg | 0.45 ± 0.03 c–f | 2.29 ± 0.09 hi | 1.48 ± 0.10 d–f | 0.24 ± 0.02 c–e | 6.17 ± 0.10 k |
| 200 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 1.11 ± 0.11 d–g | 0.44 ± 0.03 c–f | 2.52 ± 0.06 b–d | 1.55 ± 0.14 c–f | 0.24 ± 0.02 c–e | 6.46 ± 0.03 gi |
| 400 mg · L−1 ZnO + 1% Ag NPs (6% H2O) | 0.96 ± 0.07 g | 0.39 ± 0.03 f | 2.46 ± 0.02 c–g | 1.35 ± 0.10 f | 0.18 ± 0.01 g | 7.50 ± 0.16 ab |
Figure 6.
Example of RAPD (A) and SCoT (B) band profiles of the control and 400 mg · L−1 ZnO SMPs/ZnO NPs/ZnO + Ag NPs-treated plantlets of Chrysanthemum × morifolium ‘UBG’ and ‘UPG’ received as a result of electrophoresis of the DNA amplification products obtained with the R2 (RAPD) and S4 (SCoT) primers. Outermost lanes (wm) are DNA bp weight markers. NPs, nanoparticles; RAPD, randomly amplified polymorphic DNA; SCoT, start codon targeted polymorphism; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’; UPG, ‘UTP Pinky Gold’.
Table 5.
Molecular products obtained from Chrysanthemum × morifolium ‘UBG’ and ‘UPG’ control and 400 mg · L−1 ZnO SMPs/ZnO NPs/ZnO + Ag NPs-treated plantlets analysed with RAPD (R 1–5) and SCoT (S 1–5) marker systems
| Primer code | Primer sequence 5’ → 3’ | Reference | Cultivar | No. of bands | Band sizes (bp) | No. of loci | No. of genotypes | |||
|---|---|---|---|---|---|---|---|---|---|---|
| Total | mono | poly | spec | |||||||
| RAPD | ||||||||||
| R1 | GGG AAT TCG G | Lema-Rumińska et al. (2004) | UBG | 192 | 367–1,478 | 6 | 6 | 0 | 0 | 1 |
| UPG | 192 | 355–1,500 | 6 | 6 | 0 | 0 | 1 | |||
| R2 | GAC CGC TTG T | UBG | 160 | 564–1,207 | 5 | 5 | 0 | 0 | 1 | |
| UPG | 320 | 195–2,418 | 10 | 10 | 0 | 0 | 1 | |||
| R3 | GCT GCC TCA GG | Shibata et al. (1998) | UBG | 192 | 432–2,094 | 6 | 6 | 0 | 0 | 1 |
| UPG | 192 | 414–2,020 | 6 | 6 | 0 | 0 | 1 | |||
| R4 | TAC CCA GGA GCG | Wolf (1996) | UBG | 160 | 500–1,807 | 5 | 5 | 0 | 0 | 1 |
| UPG | 160 | 500–1,910 | 5 | 5 | 0 | 0 | 1 | |||
| R5 | CAA TCG CCG T | UBG | 160 | 534–1,449 | 5 | 5 | 0 | 0 | 1 | |
| UPG | 160 | 579–1,430 | 5 | 5 | 0 | 0 | 1 | |||
| Σ | UBG | 864 | 27 | 27 | 0 | 0 | 1 | |||
| UPG | 1,024 | 32 | 32 | 0 | 0 | 1 | ||||
| Mean from a single primer | UBG | 172.8 | 5.4 | 5.4 | 0 | 0 | - | |||
| UPG | 204.8 | 6.4 | 6.4 | 0 | 0 | - | ||||
| SCoT | ||||||||||
| S1 | CAA TGG CTA CCA CCT | Collard and Mackill (2009) | UBG | 352 | 425–1,762 | 11 | 11 | 0 | 0 | 1 |
| UPG | 416 | 374–1,730 | 13 | 13 | 0 | 0 | 1 | |||
| S2 | CAA TGG CTA CCA CGT | UBG | 256 | 531–1,757 | 8 | 8 | 0 | 0 | 1 | |
| UPG | 256 | 531–1,758 | 8 | 8 | 0 | 0 | 1 | |||
| S3 | ACG ACA TGG CGA CCA ACG | UBG | 352 | 509–1,888 | 11 | 11 | 0 | 0 | 1 | |
| UPG | 288 | 400–1,989 | 9 | 9 | 0 | 0 | 1 | |||
| S4 | ACG ACA TGG CGA CCA TCG | UBG | 416 | 409–2,036 | 13 | 13 | 0 | 0 | 1 | |
| UPG | 416 | 409–2,037 | 13 | 13 | 0 | 0 | 1 | |||
| S5 | ACC ATG GCT ACC GTC | UBG | 320 | 416–2,988 | 10 | 10 | 0 | 0 | 1 | |
| UPG | 256 | 329–1,824 | 8 | 8 | 0 | 0 | 1 | |||
| Σ | UBG | 1,696 | 53 | 53 | 0 | 0 | 1 | |||
| UPG | 1,632 | 51 | 51 | 0 | 0 | 1 | ||||
| Mean from a single primer | UBG | 339.2 | 10.6 | 10.6 | 0 | 0 | - | |||
| UPG | 326.4 | 10.2 | 10.2 | 0 | 0 | - | ||||
Supplementary Table S1.
Composition of the precursor solution used for synthesis
| Sample name | Ethylene glycol volume (mL) | Weight of zinc acetate dihydrate (g) | Weight of silver acetate anhydrous (g) | Final water content of the precursor solution (weight %) |
|---|---|---|---|---|
| ZnO NPs (1.5% H2O) | 0 | 1.5 | ||
| ZnO NPs (6% H2O) | 6 | |||
| ZnO + 0.1% Ag NPs (1.5% H2O) | 525 | 35 g | 0.0266 | 1.5 |
| ZnO + 0.1% Ag NPs (6% H2O) | 0.0266 | 6 | ||
| ZnO + 1% Ag NPs (1.5% H2O) | 0.2689 | 1.5 | ||
| ZnO + 1% Ag NPs (6% H2O) | 0.2689 | 6 |
Supplementary Table S2.
Characteristics of samples
| Sample name | Skeleton density, ρs ± σ (g · cm−3) | SSA, as (m2 · g−1) | Average particle size from SSA BET, d ± σ (nm) | Average crystallite size, Scherrer equation, d ± σ (nm) |
|---|---|---|---|---|
| ZnO SMPs | 5.59 ± 0.03 | 4.5 | 240 ± 30 | 124 ± 11 |
| ZnO NPs (1.5% H2O) | 5.09 ± 0.06 | 48.4 | 25 ± 2 | 31 ± 8 |
| ZnO NPs (6% H2O) | 5.38 ± 0.05 | 17.2 | 65 ± 6 | 40 ± 10 |
| ZnO + 0.1% Ag NPs (1.5% H2O) | 5.16 ± 0.07 | 40.0 | 29 ± 2 | 17 ± 5 (ZnO) |
| 31 ± 10 (Ag) | ||||
| ZnO + 0.1% Ag NPs (6% H2O) | 5.37 ± 0.06 | 14.2 | 79 ± 2 | 39 ± 12 (ZnO) |
| 31 ± 10 (Ag) | ||||
| ZnO + 1% Ag NPs (1.5% H2O) | 5.05 ± 0.05 | 44.4 | 27 ± 2 | 22 ± 3 (ZnO) |
| 45 ± 20 (Ag) | ||||
| ZnO + 1% Ag NPs (6% H2O) | 5.31 ± 0.09 | 21.4 | 53 ± 2 | 35 ± 5 (ZnO) |
| 31 ± 15 (Ag) |
Supplementary Table S3.
Results of the analysis of the chemical composition of the sample
| Sample name | Actual dopant content (mol%) | Nominal composition (mol%) | Weight content of ZnO and Ag in 100 mg of sample (mg) | |||
|---|---|---|---|---|---|---|
| Zinc | Silver | Zinc | Silver | ZnO | Ag | |
| ZnO + 0.1% Ag NPs (1.5% H2O) | 99.91 ± 0.04 | 0.09 ± 0.04 | 99.9 | 0.1 | 99.881 | 0.119 |
| ZnO + 0.1% Ag NPs (6% H2O) | 99.85 ± 0.06 | 0.15 ± 0.06 | 99.9 | 0.1 | 99.801 | 0.199 |
| ZnO + 1% Ag NPs (1.5% H2O) | 99.05 ± 0.20 | 0.95 ± 0.20 | 99.00 | 1.00 | 98.745 | 1.255 |
| ZnO + 1% Ag NPs (6% H2O) | 98.95 ± 0.20 | 1.05 ± 0.20 | 99.00 | 1.00 | 98.613 | 1.387 |
Supplementary Figure S1.
X-ray diffraction patterns of samples.
Supplementary Figure S2.
SEM images of samples: (A, B) ZnO SMPs; (C, D) ZnO NPs (1.5% H2O); (E, F) ZnO NPs (6% H2O); (G, H) ZnO + 0.1% Ag NPs (1.5% H2O); (I, J) ZnO + 0.1% Ag NPs (6% H2O); (K, L) ZnO + 1% Ag NPs (1.5% H2O); (M, N) ZnO + 1% Ag NPs (6% H2O) images taken with the immersion lens detector. NPs, nanoparticles; SMPs, submicron particles.
Supplementary Figure S3.
Chrysanthemum × morifolium ‘UBG’ and ‘UPG’ shoot and root systems developed from node explants cultured in vitro for 10 weeks on the modified MS medium, depending on the ZnO SMPs/ZnO NPs/ZnO + Ag NPs treatment; bar = 1 cm. MS, Murashige and Skoog; NPs, nanoparticles; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’; UPG, ‘UTP Pinky Gold’.
Supplementary Figure S4.
RAPD and SCoT band profiles of the control and 400 mg · L−1 ZnO SMPs/ZnO NPs/ ZnO + Ag NPs-treated plantlets of Chrysanthemum × morifolium ‘UBG’ and ‘UPG’. Outermost lanes (wm) are DNA bp weight markers. NPs, nanoparticles; RAPD, randomly amplified polymorphic DNA; SCoT, start codon targeted polymorphism; SMPs, submicron particles; UBG, ‘UTP Burgundy Gold’; UPG, ‘UTP Pinky Gold’.