Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth.
Abstract
:1. Introduction
2. Results and Discussion
2.1. Total Phenolics and Flavonoids Profiling
ABA (µM) | Parts | Total Phenolics (mg g−1 gallic acid dry weight) | Total Flavonoids (mg g−1 rutin dry weight) | Soluble sugar (mg g−1 sucrose dry weight) |
---|---|---|---|---|
Leaves | 3.11 ± 0.27 c | 1.47 ± 0.21 c | 79.12 ± 11.21 d | |
0 | Stems | 1.32 ± 0.02 l | 0.52 ± 0.02 g | 40.23 ± 8.98 l |
Roots | 2.71 ± 1.24 e | 1.21 ± 0.34 k | 62.18 ± 12.12 h | |
Leaves | 3.98 ± 0.34 b | 1.72 ± 0.56 b | 88.21 ± 9.76 c | |
2 | Stems | 1.50 ± 0.04 h | 0.76 ± 0.34 f | 47.21 ± 11.21 k |
Roots | 2.87 ± 0.45 d | 1.18 ± 0.12 j | 68.21 ± 12.12 g | |
Leaves | 4.10 ± 0.21 ab | 1.98 ± 0.32 ab | 90.17 ± 10.76 bc | |
4 | Stems | 1.57 ± 0.05 g | 0.86 ± 0.12 e | 50.11 ± 5.67 j |
Roots | 2.92 ± 0.03 d | 1.27 ± 0.32 i | 70.82 ± 5.88 f | |
Leaves | 4.21 ± 0.02 a | 2.12 ± 0.04 a | 98.12 ± 7.98 a | |
6 | Stems | 1.92 ± 0.21 f | 0.97 ± 0.08 d | 57.12 ± 12.12 l |
Roots | 2.97 ± 0.11 de | 1.46 ± 0.12 h | 76.21 ± 10.12 e |
Parameters | Pearson’s correlation coefficient (R2) | |
---|---|---|
Total phenolics | Total flavonoids | |
1. Soluble sugar | 0.971 * | 0.973 * |
2. H2O2 | 0.877 * | 0.812 * |
3. O2 | 0.778 * | 0.912 * |
4. PAL activity | 0.923 * | 0.901 * |
5. APX | 0.781 * | 0.822 ** |
6. SOD | 0.845 * | 0.816 * |
7. CAT | 0.912 * | 0.832 * |
8. ORAC | 0.904 * | 0.956 * |
9. DPPH | 0.781 * | 0.889 * |
10. Net Photosynthesis | −0.871 * | −0.921 * |
11. Stomata conductance | −0.881 ** | −0.824 * |
12. LOX | 0.951 * | 0.923 * |
2.2. Soluble Sugar Profiling
2.3. Antioxidant Content of H2O2 and O2−
ABA (µM) | Parts | H2O2 (µmol g−1 fresh weight) | O2− (µmol g−1 dry weight min−1) | PAL Activity (nm transcinnamic mg−1 protein−1 h−1) |
---|---|---|---|---|
Leaves | 2.19 ± 0.34 d | 1.09 ± 0.02 d | 9.21 ± 0.62 d | |
0 | Stems | 0.67 ± 0.21 k | 0.32 ± 0.23 j | 2.08 ± 0.23 l |
Roots | 1.32 ± 0.23 h | 0.66 ± 0.45 g | 5.02 ± 0.15 h | |
Leaves | 2.43 ± 0.31 c | 1.17 ± 0.32 c | 10.16 ± 0.82 c | |
2 | Stems | 0.78 ± 0.03 k | 0.44 ± 0.12 i | 2.17 ± 0.12 k |
Roots | 1.52 ± 0.34 g | 0.72 ± 0.02 f | 6.23 ± 0.32 g | |
Leaves | 2.60 ± 0.06 b | 1.23 ± 0.12 b | 13.11 ± 2.13 b | |
4 | Stems | 0.90 ± 0.02 j | 0.56 ± 0.13 h | 3.21 ± 0.19 j |
Roots | 1.66 ± 0.21 f | 0.87 ± 0.01 e | 7.11 ± 0.34 f | |
Leaves | 2.81 ± 0.06 a | 1.57 ± 0.21 a | 17.21 ± 1.21 a | |
6 | Stems | 0.91 ± 0.12 i | 0.57 ± 0.11 h | 4.11 ± 0.6l i |
Roots | 1.78 ± 0.02 e | 0.91 ± 0.04 e | 8.81 ± 0.54 e |
2.4. Phenyl Alanine Ammonia Lyase (PAL) Activity
2.5. Antioxidant Enzyme Activities
ABA (µM) | Parts | Ascorbate peroxidase activity (APX; mg protein−1 min−1) | Superoxide dismutase activity (SOD mg protein−1 min−1) | Catalase activity (CAT; µmol mg protein−1 min−1) |
---|---|---|---|---|
Leaves | 15.23 ± 2.34 d | 4.62 ± 0.11 d | 19.21 ± 1.27 d | |
0 | Stems | 6.12 ± 0.81 k | 1.34 ± 0.01 l | 6.66 ± 2.11 l |
Roots | 10.11 ± 0.03 h | 2.98 ± 0.41 h | 12.17 ± 0.97 h | |
Leaves | 17.11 ± 0.51 c | 4.82 ± 0.21 c | 20.12 ± 0.82 c | |
2 | Stems | 6.11 ± 0.53 k | 1.52 ± 0.36 k | 8.27 ± 0.78 k |
Roots | 11.27 ± 0.14 g | 3.62 ± 0.15 g | 13.24 ± 0.11 g | |
Leaves | 19.71 ± 0.16 b | 5.01 ± 0.17 b | 23.17 ± 0.78 b | |
4 | Stems | 7.23 ± 0.42 j | 1.71 ± 2.11 j | 9.23 ± 1.19 j |
Roots | 13.22 ± 0.31 f | 3.89 ± 1.02 f | 16.59 ± 0.89 f | |
Leaves | 21.62 ± 0.26 a | 5.27 ± 0.81 a | 25.12 ± 1.21 a | |
6 | Stems | 9.12 ± 0.98 i | 1.76 ± 0.92 i | 10.24 ± 2.17 i |
Roots | 14.21 ± 1.32 e | 4.02 ± 1.24 e | 17.21 ± 0.98 e |
2.6. 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) and Oxygen Radical Absorbance Capacity (ORAC) Assay
ABA (µM) | Parts | ORAC (µmol Trolox equivalent g−1) | DPPH (µmol Trolox equivalent g−1) |
---|---|---|---|
Leaves | 65.21 ± 2.41 d | 20.19 ± 9.02 d | |
0 | Stems | 35.67 ± 0.21 i | 7.72 ± 2.32 j |
Roots | 51.07 ± 2.23 g | 13.66 ± 6.45 h | |
Leaves | 69.31 ± 10.31 c | 25.17 ± 8.32 c | |
2 | Stems | 37.78 ± 9.03 k | 9.44 ± 7.12 k |
Roots | 55.02 ± 0.34 l | 15.72 ± 3.02 g | |
Leaves | 70.60 ± 8.96 b | 27.84 ± 9.12 b | |
4 | Stems | 40.90 ± 11.02 h | 10.23 ± 8.13 i |
Roots | 57.21 ± 10.21 f | 16.21 ± 2.21 f | |
Leaves | 77.81 ± 0.06 a | 31.57 ± 9.21 a | |
6 | Stems | 45.21 ± 0.12l g | 11.01 ± 2.11 h |
Roots | 60.23 ± 8.02 e | 17.81 ± 2.04 e |
2.7. Leaf Gas Exchange
2.8. Lipoxygenase (LOX) Inhibitory Activity
3. Experimental
3.1. Plant Materials and Maintenance
Microclimate parameters | Quantification |
---|---|
Relative Humidity | 56.14–65.32% |
Light intensity | 225–1450 µmol m−2s−1 |
Day temperature | 27–31 °C |
Night temperature | 18–22 °C |
Ambient CO2 | 372.81 µmol mol−1 |
3.2. Determination of Total Phenolics and Flavonoids
3.3. Determination of Soluble Sugar
3.4. Superoxide Radical (O2−) Assay
3.5. H2O2 Assay
3.6. Oxygen Radical Absorbance Capacity (ORAC) Assay
3.7. DPPH Assay
3.8. Phenylalanine Ammonia-Lyase (PAL) Activity
3.9. Leaf Gas Exchange Measurement
3.10. Antioxidant Enzyme Activity
3.10.1. Preparation of Enzyme Extracts
3.10.2. Ascorbate Peroxidase (APX) Activity Assay
3.10.3. Catalase (CAT) Activity Assay
3.10.4. Superoxide Dismutase (SOD) Activity Assay
3.11. LOX Inhibitory Assay
3.12. Statistical Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Ibrahim, M.H.; Jaafar, H.Z.E. Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth. Molecules 2013, 18, 7957-7976. https://doi.org/10.3390/molecules18077957
Ibrahim MH, Jaafar HZE. Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth. Molecules. 2013; 18(7):7957-7976. https://doi.org/10.3390/molecules18077957
Chicago/Turabian StyleIbrahim, Mohd Hafiz, and Hawa Z. E. Jaafar. 2013. "Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth." Molecules 18, no. 7: 7957-7976. https://doi.org/10.3390/molecules18077957