The Influence of Nasturtium officinale R. Br. Agar and Agitated Microshoot Culture Media on Glucosinolate and Phenolic Acid Production, and Antioxidant Activity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Parent Plant Material
2.2. Initiation of In Vitro Cultures
2.3. Experimental In Vitro Cultures
2.3.1. Agar Microshoot Cultures
2.3.2. Agitated Microshoot Cultures
2.4. Biomass Growth
2.5. Spectrophotometric Analysis of the Total Glucosinolate Pool
2.6. Phenolic acid Analysis
2.7. Total Phenolic Assay
2.8. Antioxidant Capacity
2.8.1. CUPRAC Total Antioxidant Capacity Assay
2.8.2. Ferric Reducing Ability (FRAP) Assay
2.8.3. 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) Radical-Scavenging Activity Assay
2.9. Statistical Model Fitting
3. Results
3.1. The Experimental In Vitro Cultures
3.1.1. Microshoot Appearance and Biomass Growth
Agar Microshoot Cultures
Agitated Microshoot Cultures
Statistical Analysis
3.1.2. Effect of Supplementation with Different PGRs on Glucosinolate Production
Agar Microshoot Cultures
Agitated Microshoot Cultures
Statistical Analysis
3.1.3. Effect of PGRs on Phenolic Acid Production
Agar Microshoot Cultures
Agitated Microshoot Cultures
Statistical Analysis
- The amounts of gallic, protocatechuic, caffeic, and syringic acids, and the total amount of phenolic acids were significantly higher in the agitated cultures under the same conditions.
- Additions of BA, NAA, Zea, IAA, 2iP, KIN, 2,4-D and IBA caused an increase in many compounds and sometimes a decrease in p-coumaric, o-coumaric, or ferulic acids. This change was correlated with an opposite change of the slope against time (when average content is higher, production in time is lower, and vice versa). For the agitated cultures, the addition of NAA did not much increase the synthesis of gallic, protocatechuic, caffeic, and syringic acids, or the total amount of phenolic acids.
- There were several significant interactions among the added compounds, where the synthesis of the compounds was significantly lower than one could expect from the additivity of positive effects.
3.1.4. The Effect of PGRs on the Antioxidant Potential and Total Phenolic Content of Biomass
Agar Microshoot Cultures
Agitated Microshoot Cultures
Statistical Analysis
3.2. Parent Plant Material
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Type of Microshoot Cultures | Growth Cycles (days) | Min Gi | Max Gi | Increase (fold) | MS Medium Variants |
---|---|---|---|---|---|
Agar | 10 | 0.05 ± 0.01 | 1.75 ± 0.03 | 35.00 | 1 mg/L BA and 1 mg/L NAA |
20 | 0.37 ± 0.04 | 3.79 ± 0.05 | 10.24 | ||
30 | 1.28 ± 0.04 | 5.05 ± 0.14 | 3.95 | ||
Agitated | 10 | 1.47 ± 0.02 | 5.29 ± 0.07 | 3.60 | 1 mg/L BA and 1 mg/L NAA1 1 mg/L KIN and 1 mg/L IBA |
20 | 1.59 ± 0.01 | 10.48 ± 0.08 | 6.59 | 1 mg/L Zea and 1 mg/L NAA |
Phenolic Acids | Min Content | Max Content | Increase (fold) | Conditions for Max Content | ||
---|---|---|---|---|---|---|
Growth Cycle (days) | MS Medium Variant | Gi | ||||
Caffeic acid | 0.04 ± 0.01 | 16.65 ± 1.59 | 416.25 | 20 | 2 mg/L BA and 1 mg/L NAA | 2.96 ± 0.06 |
o-Coumaric acid | 0.03 ± 0.01 | 17.34 ± 2.09 | 578.00 | 30 | 1 mg/L 2iP and 1 mg/L IPA | 2.06 ± 0.03 |
p-Coumaric acid | 2.14 ± 0.18 | 18.95 ± 2.11 | 8.86 | 20 | 1 mg/L KIN and 1 mg/L IPA | 0.42 ± 0.01 |
Ellagic acid | 2.79 ± 0.25 | 6.56 ± 0.74 | 2.35 | 30 | 1 mg/L 2iP | 1.59 ± 0.05 |
Ferulic acid | 1.64 ± 0.20 | 18.13 ± 1.13 | 11.05 | 10 | 1 mg/L KIN and 1 mg/L IAA | 0.30 ± 0.02 |
Gallic acid | 1.40 ± 0.13 | 61.03 ± 5.89 | 43.59 | 30 | 2 mg/l BA and 1 mg/L NAA | 2.95 ± 0.04 |
Isoferulic acid | 0.02 ± 0.01 | 6.01 ± 0.59 | 300.50 | 10 | 1 mg/L KIN and 1 mg/L IBA | 0.23 ± 0.03 |
Protocatechuic acid | 0.78 ± 0.08 | 138.40 ± 12.99 | 177.44 | 20 | 2 mg/L BA and 1 mg/L NAA | 2.96 ± 0.06 |
Rosmarinic acid | 0.07 ± 0.01 | 33.30 ± 3.67 | 475.71 | 20 | 1 mg/L 2iP and 1 mg/L NAA | 1.59 ± 0.01 |
Syringic acid | 1.65 ± 0.18 | 9.65 ± 0.89 | 5.85 | 20 | 1 mg/L BA and 1 mg/L NAA | 3.79 ± 0.05 |
Total content | 15.89 ± 1.66 | 237.52 ± 22.56 | 14.95 | 20 | 2 mg/L BA and 1 mg/L NAA | 2.96 ± 0.06 |
Phenolic Acids | Min Content | Max Content | Increase (fold) | Conditions for Max Content | ||
---|---|---|---|---|---|---|
Growth Cycle (days) | MS Medium Variant | Gi | ||||
Caffeic acid | 0.02 ± 0.01 | 15.30 ± 1.66 | 765.00 | 10 | 1 mg/L Zea and 1 mg/L NAA | 1.75 ± 0.01 |
o-Coumaric acid | 0.74 ± 0.66 | 14.91 ± 1.35 | 20.15 | 10 | 1 mg/L BA and 1 mg/L NAA | 5.29 ± 0.07 |
p-Coumaric acid | 4.84 ± 0.45 | 31.66 ± 2.98 | 6.54 | 10 | 1 mg/L BA and 1 mg/L NAA | 5.29 ± 0.07 |
Ellagic acid | 2.93 ± 0.33 | 8.12 ± 0.79 | 2.69 | 20 | 1 mg/L KIN and 1 mg/L IBA | 7.51 ± 0.03 |
Ferulic acid | 1.58 ± 1.64 | 38.44 ± 4.12 | 24.33 | 10 | 1 mg/L BA and 1 mg/L NAA | 5.29 ± 0.07 |
Gallic acid | 29.84 ± 0.28 | 53.34 ± 5.12 | 1.79 | 10 | 1 mg/L Zea and 1 mg/L NAA | 1.75 ± 0.01 |
Isoferulic acid | 0.02 ± 0.01 | 15.17 ± 1.49 | 758.50 | 10 | 1 mg/L BA and 1 mg/L NAA | 5.29 ± 0.07 |
Protocatechuic acid | 4.94 ± 0.48 | 132.26 ± 12.02 | 26.77 | 10 | 1 mg/L Zea and 1 mg/L NAA | 1.75 ± 0.01 |
Rosmarinic acid | 2.95 ± 0.33 | 36.34 ± 3.71 | 12.32 | 20 | 1 mg/L 2iP and 1 mg/L NAA | 1.59 ± 0.01 |
Syringic acid | 1.65 ± 0.18 | 21.50 ± 2.12 | 13.03 | 10 | 1 mg/L BA and 1 mg/L NAA | 5.29 ± 0.07 |
Total content | 70.80 ± 7.12 | 236.74 ± 19.30 | 3.34 | 10 | 1 mg/L Zea and 1 mg/L NAA | 1.75 ± 0.01 |
Culture Type | Method | Min (mmol TE/100 g DW ± SD) | Max (mmol TE/100 g DW ± SD) | Increase (fold) | Conditions for Max Antioxidant Parameters | ||
---|---|---|---|---|---|---|---|
Growth Cycle (days) | MS Medium Variant | Gi | |||||
Agar | CUPRAC | 1.50 ± 0.19 | 4.13 ± 0.80 | 1.69 | 30 | 1 mg/L KIN and 1 mg/L IAA | 2.39 ± 0.05 |
FRAP | 0.52 ± 0.02 | 1.42 ± 0.02 | 2.73 | 30 | 2 mg/L BA and 1 mg/L NAA | 2.95 ± 0.04 | |
DPPH | 0.49 ± 0.04 | 30.89 ± 1.82 | 63.04 | 30 | 1 mg/L KIN and 1 mg/L IAA | 2.39 ± 0.05 | |
F-C | 2.42 ± 0.10 | 8.69 ± 3.44 | 3.59 | 10 | 1 mg/L Zea and 1 mg/L NAA | 3.27 ± 0.05 | |
Agitated | CUPRAC | 2.52 ± 0.15 | 5.26 ± 0.10 | 2.09 | 20 | 1 mg/L KIN and 1 mg/L IAA | 8.08 ± 0.06 |
FRAP | 0.51 ± 0.04 | 1.26 ± 0.04 | 2.47 | 20 | 1 mg/L KIN and 1 mg/L IAA | 8.08 ± 0.06 | |
DPPH | 14.55 ± 0.96 | 25.95 ± 4.07 | 1.78 | 20 | 1 mg/L KIN and 1 mg/L IAA | 8.08 ± 0.06 | |
F-C | 2.49 ± 0.13 | 5.24 ± 0.20 | 2.10 | 20 | 1 mg/L KIN and 1 mg/L IAA | 8.08 ± 0.06 |
Total Glucosinolate Content (mg/100g DW ± SD) | Phenolic Acid Contents (mg/100g DW ± SD) | Antioxidant Activity | ||
---|---|---|---|---|
Method | (mmol TE/100 g DW ± SD) | |||
499.89 ± 34.59 | Caffeic acid | 7.10 ± 0.65 | CUPRAC | 4.45 ± 0.02 |
o-Coumaric acid | 2.39 ± 0.19 | |||
p-Coumaric acid | 0.61 ± 0.05 | FRAP | 0.76 ± 0.08 | |
Ferulic acid | 14.32 ± 1.43 | |||
Gallic acid | 13.67 ± 1.12 | DPPH | 26.32 ± 8.23 | |
Rosmarinic acid | 61.27 ± 5.87 | |||
Syringic acid | 0.48 ± 0.05 | F-C | 2.70 ± 0.31 | |
Total content | 99.84 ± 9.65 |
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Klimek-Szczykutowicz, M.; Szopa, A.; Dziurka, M.; Komsta, Ł.; Tomczyk, M.; Ekiert, H. The Influence of Nasturtium officinale R. Br. Agar and Agitated Microshoot Culture Media on Glucosinolate and Phenolic Acid Production, and Antioxidant Activity. Biomolecules 2020, 10, 1216. https://doi.org/10.3390/biom10091216
Klimek-Szczykutowicz M, Szopa A, Dziurka M, Komsta Ł, Tomczyk M, Ekiert H. The Influence of Nasturtium officinale R. Br. Agar and Agitated Microshoot Culture Media on Glucosinolate and Phenolic Acid Production, and Antioxidant Activity. Biomolecules. 2020; 10(9):1216. https://doi.org/10.3390/biom10091216
Chicago/Turabian StyleKlimek-Szczykutowicz, Marta, Agnieszka Szopa, Michał Dziurka, Łukasz Komsta, Michał Tomczyk, and Halina Ekiert. 2020. "The Influence of Nasturtium officinale R. Br. Agar and Agitated Microshoot Culture Media on Glucosinolate and Phenolic Acid Production, and Antioxidant Activity" Biomolecules 10, no. 9: 1216. https://doi.org/10.3390/biom10091216