Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L.
Abstract
:1. Introduction
2. Results
2.1. Stabilization of Shoot Cultures Agitated in Liquid Media
2.2. Effect of (+)-Usnic Acid on Phenolic Compound Content in Shoot Cultures
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Shoot Cultures Initiation
4.3. Shoot Cultures Agitated in Liquid Media
4.4. Elicitation Process
4.5. HPLC-DAD Analyses
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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PGRs (mg/L). | Induction | Shoot No./Explant | Shoot Length | ||
---|---|---|---|---|---|
BAP | IAA | GA3 | [%] | No. ± SD | [cm] |
0.0 | 0.0 | 0.0 | 100 n,s | 2.67 ± 0.58 b | 4.53 ± 0.31 b |
1.0 | 1.0 | 0.0 | 100 | 8.33 ± 1.53 a | 5.67 ± 1.04 b |
1.0 | 1.0 | 1.0 | 100 | 9.00 ± 1.00 a | 7.60 ± 0.66 a |
1.0 | 1.0 | 1.5 | 100 | 7.66 ± 0.58 a | 7.93 ± 0.59 a |
1.0 | 1.0 | 2.0 | 100 | 8.67 ± 0.58 a | 8.03 ± 1.17 a |
1.0 | 1.0 | 2.5 | 100 | 8.33 ± 1.53 a | 7.77 ± 0.42 a |
1.0 | 1.0 | 3.0 | 100 | 9.33 ± 1.15 a | 7.67 ± 0.29 a |
Treatment | Flavonoids (mg/100 g DW ±SD) | ||||
---|---|---|---|---|---|
Conc. | Time | Isoquercetin | Robinin | Quercitrin | Total |
0 | 0 | 82.10 ± 0.62 c,d | 15.39 ± 0.23 c,d,e | 35.11 ± 0.19 d,e,f | 132.60 |
3.125 µM | 12 h | 70.97 ± 0.17 ef | 18.13 ± 0.44 b,c,d | 29.56 ± 0.13 I,j | 118.66 |
3.125 µM | 24 h | 93.87 ± 1.49 b | 37.32 ± 2.59 a | 45.88 ± 0.98 b | 177.07 |
3.125 µM | 48 h | 24.12 ± 5.09 j | 9.66 ± 3.84 f,g | 42.94 ± 0.34 b | 76.72 |
3.125 µM | 72 h | 107.17 ± 4.67 a | 17.88 ± 0.07 b,c,d | 51.08 ± 0.17 a | 176.13 |
6.25 µM | 12 h | 67.43 ± 4.19 f,g | 16.38 ± 2.09 b,c,d,e | 28.44 ± 0.46 j | 112.25 |
6.25 µM | 24 h | 62.38 ± 0.48 g,h | 11.97 ± 0.25 e,f | 29.10 ± 0.36 i,j | 103.45 |
6.25 µM | 48 h | 76.04 ± 0.63 d,e | 17.36 ± 0.79 b,c,d | 33.72 ± 0.75 f,g,h | 127.12 |
6.25 µM | 72 h | 58.83 ± 0.46 h | 6.02 ± 1.11 g,h | 32.02 ± 0.25 f,g,h,i | 96.87 |
12.5 µM | 12 h | 127.54 ± 11.34 a | 15.32 ± 6.62 c,d,e | 35.13 ± 3.01 def | 177.99 |
12.5 µM | 24 h | 93.93 ± 0.31 b | 19.01 ± 0.51 b,c | 32.05 ± 0.03 f,g,h,i | 144.99 |
12.5 µM | 48 h | 73.81 ± 0.63 e,f | 13.93 ± 0.02 d,e,f | 34.81 ± 0.05 d,e,f,g | 122.55 |
12.5 µM | 72 h | 89.55 ± 5.68 b,c | 6.12 ± 0.45 g,h | 35.19 ± 0.69 d,e,f | 130.86 |
25 µM | 12 h | 108.37 ± 12.1 a | 20.67 ± 6.40 b | 38.78 ± 7.21 c | 167.82 |
25 µM | 24 h | 89.23 ± 0.61 b,c | 15.78 ± 0.22 b,c,d,e | 31.32 ± 0.06 h,I,j | 136.33 |
25 µM | 48 h | 85.77 ± 3.63 c | 12.34 ± 0.66 e,f | 34.08 ± 0.67 e,f,g,h | 132.19 |
25 µM | 72 h | 62.41 ± 0.33 g,h | 5.00 ± 0.12 h | 37.28 ± 0.55 c,d,e | 104.69 |
50 µM | 12 h | 71.93 ± 1.16 e,f | 10.35 ± 0.71 f,g | 31.59 ± 0.52 g,h,i,j | 113.87 |
50 µM | 24 h | 31.22 ± 1.53 j | 3.77 ± 0.31 i | 28.89 ± 0.13 i,j | 63.88 |
50 µM | 48 h | 106.74 ± 3.35 a | 17.63 ± 4.27 b,c,d | 38.09 ± 0.84 c,d | 162.46 |
50 µM | 72 h | 41.36 ± 0.14 i | 4.04 ± 0.14 i | 34.88 ± 0.04 d,e,f,g | 80.28 |
Treatment | Phenolic Acids (mg/100 g DW ± SD) | ||||||||
---|---|---|---|---|---|---|---|---|---|
Conc. | Time | Caftaric Acid | 3,4-Dihydroxy-phenylacetic Acid | Chlorogenic Acid | Cryptochlorogenic Acid | Isochlorogenic Acid | Caffeic Acid | Rosmarinic Acid | Total |
0 | 0 | 26.82 ± 0.14 g,h | 116.40 ± 2.51 h,i | 14.81 ± 0.79 e | 55.18 ± 0.47 a | 34.52 ± 1.79 b | 165.60 ± 0.77 d | 245.27 ± 0.89 e | 658.60 |
3.125 µM | 12 h | 26.34 ± 0.65 h | 191.14 ± 10.95 b | 6.75 ± 1.03 j | 31.89 ± 5.39 f,g | 3.81 ± 0.23 g | 51.09 ± 6.46 j | 204.30 ± 3.38 f | 515.32 |
3.125 µM | 24 h | 43.32 ± 1.11 a | 182.17 ± 4.48 b,c | 12.11 ± 1.35 f,g | 50.73 ± 6.11 a,b | 41.04 ± 0.51 a | 200.83 ± 11.93 b | 512.69 ± 4.89 a | 1042.89 |
3.125 µM | 48 h | 7.17 ± 0.20 m | 26.95 ± 0.69 l | 4.28 ± 0.22 k | 14.73 ± 0.04 i | 13.45 ± 4.09 d,e | 20.98 ± 2.09 k | 79.21 ± 4.55 i | 166.77 |
3.125 µM | 72 h | 32.48 ± 0.88 d | 157.38 ± 5.46 e,f,g | 14.59 ± 1.71 e | 46.80 ± 0.86 b,c | 21.13 ± 0.83 c | 178.93 ± 7.41 c | 361.70 ± 21.23 c | 813.01 |
6.25 µM | 12 h | 37.31 ± 1.58 b | 174.54 ± 9.51 c,d | 8.96 ± 0.36 h,I,j | 39.53 ± 8.05 d,e | 5.93 ± 1.82 f,g | 68.47 ± 4.04 i | 216.75 ± 12.79 f | 551.49 |
6.25 µM | 24 h | 27.61 ± 0.37 f,g,h | 151.59 ± 1.83 e,f,g | 9.98 ± 0.66 g,h,i | 41.91 ± 2.00 c,d | 6.14 ± 0.79 f,g | 100.11 ± 2.31 g | 182.47 ± 1.69 f | 519.81 |
6.25 µM | 48 h | 19.98 ± 0.35 j | 64.71 ± 2.03 k | 22.13 ± 0.36 d | 35.62 ± 0.84 e,f | 15.45 ± 0.45 d | 79.39 ± 1.79 h | 130.49 ± 1.60 h | 367.77 |
6.25 µM | 72 h | 22.21 ± 1.47 i | 125.19 ± 10.97 h | 9.91 ± 0.56 g,h,i | 27.84 ± 5.32 g,h | 10.43 ± 3.12 d,e,f | 118.59 ± 2.95 f | 263.02 ± 0.76 e | 577.19 |
12.5 µM | 12 h | 32.84 ± 0.33 d | 162.91 ± 2.49 d,e,f | 10.39 ± 0.00 g,h | 24.21 ± 0.87 h | 7.96 ± 3.06 e,f,g | 69.30 ± 1.88 i | 237.09 ± 15.24 e | 544.70 |
12.5 µM | 24 h | 35.59 ± 0.80 c | 166.64 ± 1.36 d,e | 9.64 ± 0.61 g,h,i | 47.47 ± 3.23 b,c | 13.05 ± 0.36 d,e | 126.26 ± 4.74 f | 356.09 ± 2.07 c | 754.74 |
12.5 µM | 48 h | 29.39 ± 0.67 e | 171.58 ± 0.78 c,d,e | 31.29 ± 1.09 b | 0.00 ± 0.00 j | 22.22 ± 0.27 c | 158.20 ± 1.31 d,e | 269.52 ± 1.92 e | 682.20 |
12.5 µM | 72 h | 28.39 ± 0.35 e,f | 161.64 ± 5.23 d,e,f,g | 27.71 ± 1.78 c | 39.96 ± 5.00 d,e | 2.44 ± 0.73 g | 175.51 ± 9.26 c | 393.93 ± 23.66 b,c | 829.58 |
25 µM | 12 h | 31.58 ± 1.09 d | 205.47 ± 17.02 a | 7.73 ± 0.75 i,j | 26.19 ± 0.72 h | 23.46 ± 10.50 c | 94.17 ± 7.44 g | 275.12 ± 26.71 e | 663.72 |
25 µM | 24 h | 34.63 ± 0.43 c | 139.77 ± 22.30 g | 11.85 ± 3.52 g | 49.49 ± 1.18 b | 8.44 ± 0.72 e,f,g | 119.73 ± 4.95 f | 282.17 ± 2.05 e | 646.08 |
25 µM | 48 h | 28.03 ± 1.13 e,f,g | 98.89 ± 3.76 j | 32.47 ± 1.48 b | 0.00 ± 0.00 j | 23.21 ± 1.43 c | 164.73 ± 7.20 d | 245.76 ± 10.57 e | 593.09 |
25 µM | 72 h | 22.27 ± 0.17 i | 148.21 ± 0.33 f,g | 10.32 ± 0.54 gh | 34.65 ± 0.83 e,f | 15.06 ± 0.45 d | 217.78 ± 1.84 a | 412.74 ± 1.58 b | 861.03 |
50 µM | 12 h | 27.34 ± 0.55 f,g,h | 160.15 ± 3.41 d,e,f,g | 23.89 ± 0.49 d | 22.14 ± 0.57 h | 5.53 ± 1.38 f,g | 118.57 ± 1.69 f | 326.16 ± 2.91 d | 683.78 |
50 µM | 24 h | 18.52 ± 0.78 k | 107.26 ± 3.78 i,j | 16.59 ± 1.42 e | 0.00 ± 0.00 j | 10.92 ± 3.59 d,e,f | 62.97 ± 2.58 i | 185.15 ± 4.29 g | 401.41 |
50 µM | 48 h | 29.10 ± 0.53 e | 125.27 ± 0.95 h | 43.44 ± 0.33 a | 0.00 ± 0.00 j | 33.25 ± 7.86 b | 218.38 ± 0.95 a | 401.66 ± 22.18 b | 851.10 |
50 µM | 72 h | 16.29 ± 0.46 l | 118.31 ± 2.73 h,i | 14.32 ± 3.22 e,f | 25.57 ± 4.23 h | 11.87 ± 0.33 d,e,f | 155.46 ± 2.91 e | 263.08 ± 0.76 e | 604.90 |
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Kikowska, M.; Thiem, B.; Jafernik, K.; Klimek-Szczykutowicz, M.; Studzińska-Sroka, E.; Ekiert, H.; Szopa, A. Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L. Molecules 2021, 26, 5532. https://doi.org/10.3390/molecules26185532
Kikowska M, Thiem B, Jafernik K, Klimek-Szczykutowicz M, Studzińska-Sroka E, Ekiert H, Szopa A. Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L. Molecules. 2021; 26(18):5532. https://doi.org/10.3390/molecules26185532
Chicago/Turabian StyleKikowska, Małgorzata, Barbara Thiem, Karolina Jafernik, Marta Klimek-Szczykutowicz, Elżbieta Studzińska-Sroka, Halina Ekiert, and Agnieszka Szopa. 2021. "Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L." Molecules 26, no. 18: 5532. https://doi.org/10.3390/molecules26185532