Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant
Abstract
:1. Introduction
2. Results
2.1. Shoot Multiplication
2.2. Callus Induction
2.2.1. Callus Induction Using a Single Concentration of 2,4-D and Picloram
2.2.2. Callus Induction using Combination of 0.5 mg/L 2,4-D and 0.25 mg/L Different Cytokinins
2.2.3. Callus Induction Using a Combination of 1.0 mg/L Picloram and 0.5 mg/L Cytokinins
2.3. Total Antioxidant, Phenolic, and Flavonoid Content in Wild Plant and In Vitro Culture of Labisia pumila var. alata
2.3.1. Total Phenolic and Flavonoid Content in Wild Plant, In Vitro-Derived Plantlet, and Callus of Labisia pumila var. alata
2.3.2. Total Antioxidant (DPPH and FRAP Methods) in Wild Plant, In Vitro-Derived Plantlet, and Callus of Labisia pumila var. alata
2.3.3. Correlation between Antioxidant Activities and Total Phenolic and Flavonoid Content in Wild Plant, In Vitro-Derived Plantlet, and Callus of Labisia pumila var. alata
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Maintenance
4.2. Media Preparation
4.3. Shoot Multiplication
4.4. Callus Induction and Maintenance
4.5. Measurement of Total Antioxidant Activity
4.5.1. Extract Preparation
4.5.2. Ferric Reducing Antioxidant Potential Assay (FRAP)
4.5.3. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Free Radical Scavenging Assay
4.5.4. Total Phenolic Content Determination
4.5.5. Total Flavonoid Content Determination
4.6. Experimental Design and Data Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Growth Regulators (mg/L) | No. of Shoots | Length of Shoot (cm) | No. of Leaves | ||
---|---|---|---|---|---|
BAP | Kin | Zea | |||
0 | 0 | 0 | 5.56 abcd | 3.89 ab | 2.1 bc |
1 | 0 | 0 | 6.15 abc | 3.55 abc | 1.6 bcdef |
2 | 0 | 0 | 2.03 g | 2.15 efg | 1.0 def |
3 | 0 | 0 | 3.92 bcdef | 2.51 defg | 1.73 bcde |
4 | 0 | 0 | 4.06 abcdef | 2.49 defg | 1.4 cdef |
5 | 0 | 0 | 3.25 efg | 2.37 defg | 1.42 cdef |
0 | 0 | 0 | 4.19 abcdef | 3.19 abcd | 1.8 bcd |
0 | 1 | 0 | 3.32 efg | 2.61 cdefg | 0.93 ef |
0 | 2 | 0 | 3.13 fg | 2.54 defg | 0.93 ef |
0 | 3 | 0 | 3.61 def | 2.26 efg | 1.1 def |
0 | 4 | 0 | 3.75 cdef | 2.71 cdefg | 1.4 cdef |
0 | 5 | 0 | 2.12 g | 2.13 fg | 0.8 f |
0 | 0 | 0 | 2.74 fg | 2.10 g | 1.4 cdef |
0 | 0 | 1 | 5.36 abcde | 4.32 a | 2.4 b |
0 | 0 | 2 | 6.53 ab | 3.50 abc | 3.13 a |
0 | 0 | 3 | 6.64 a | 2.95 bcde | 2.27 b |
0 | 0 | 4 | 5.29 abcde | 2.93 bcdef | 1.0 def |
0 | 0 | 5 | 6.10 abc | 2.89 bcdefg | 1.0 def |
Growth Regulators (mg/L) | Callus Induction Percentage (%) | Callus Score | Morphology, Texture | |
---|---|---|---|---|
2,4-D | Picloram | |||
0 | 0 | NC | - | NC |
0.5 | 0 | 60 a | ++ | Greenish, compact |
1 | 0 | 6.7 c | ++ | Greenish, compact |
1.5 | 0 | 6.7 c | + | Friable, yellowish to greenish |
2 | 0 | NC | - | NC |
2.5 | 0 | NC | - | NC |
0 | 0.5 | 6.7 c | + | Friable, yellowish |
0 | 1 | 50 a | +++ | Friable, whitish |
0 | 1.5 | 30 b | ++ | Friable, whitish |
0 | 2 | 27 b | ++ | Friable, yellowish to whitish |
0 | 2.5 | 27 b | + | Friable, yellowish to whitish |
Growth Regulators (mg/L) | Callus Formation (%) | Callus Score | Morphology, Texture |
---|---|---|---|
0.5 2,4-D + 0.25 zeatin | 100 a | +++ | Compact, yellowish to greenish |
0.5 2,4-D + 0.25 kinetin | 73.4 ab | ++ | Friable, yellowish to greenish |
0.5 2,4-D + 0.25 BAP | 46.6 b | + | Friable, yellowish to greenish |
0.5 2,4-D + 0.25 TDZ | 6.6 c | + | Friable, whitish |
Growth Regulators (Mg/L) | Callus Formation Percentage (%) | Callus Score | Morphology, Texture |
---|---|---|---|
1.0 picloram + 0.5 zeatin | 33.4 ab | + | Greenish, compact |
1.0 picloram + 0.5 kinetin | 80 a | +++ | Whitish, friable |
1.0 picloram + 0.5 BAP | 66.6 ab | ++ | Whitish, compact |
1.0 picloram + 0.5 TDZ | 20 b | + | Whitish to dark brown, compact |
Source of Sample | Total Phenolic Content (mg GAE/g DW) | Total Flavonoid Content (mg QE/g DW) |
---|---|---|
Wild leaves | 1.01 ± 0.07 c | 1.67 ± 0.02 c |
In vitro plantlets | 1.4 ± 0.07 b | 1.41 ± 0.68 b |
Callus | 1.9 ± 0.14 a | 2.38 ± 0.18 a |
Total Antioxidant Content (mg TE/g DW) | ||
---|---|---|
Source of Sample | DPPH | FRAP |
Wild plant | 40.45 ± 2.69 c | 81 ± 5.39 c |
In vitro plantlet | 57.47 ± 2.63 b | 115 ± 5.25 b |
Callus | 75.88 ± 5.80 a | 152 ± 11.59 a |
Variables | 1 | 2 | 3 | 4 | |
---|---|---|---|---|---|
1 | TPC | 1 | |||
2 | TFC | 0.86 * | 1 | ||
3 | DPPH | 0.81 * | 0.91 * | 1 | |
4 | FRAP | 0.98 * | 0.91 * | 0.86 * | 1 |
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Najhah, M.Y.; Jaafar, H.Z.E.; Nakasha, J.J.; Hakiman, M. Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant. Molecules 2021, 26, 3229. https://doi.org/10.3390/molecules26113229
Najhah MY, Jaafar HZE, Nakasha JJ, Hakiman M. Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant. Molecules. 2021; 26(11):3229. https://doi.org/10.3390/molecules26113229
Chicago/Turabian StyleNajhah, Mat Yunus, Hawa Z. E. Jaafar, Jaafar Juju Nakasha, and Mansor Hakiman. 2021. "Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant" Molecules 26, no. 11: 3229. https://doi.org/10.3390/molecules26113229