Enhanced Antioxidant and Anticancer Potential of Artemisia carvifolia Buch Transformed with rol A Gene
Abstract
:1. Introduction
2. Materials and Methods
2.1. Seed Germination and DNA Barcoding
2.2. Bacterial Strains and Plasmids
2.3. Transformation and Regeneration
2.4. Molecular Analysis
2.5. Southern Blotting
2.6. Evaluation of Flavonoids through an HPLC-DAD System
2.7. Analysis of Genes Involved in the Flavonoid Biosynthetic Pathway Using Real-Time qPCR
2.8. Antioxidant Potential Measurement
2.8.1. Total Phenolic Content Measurement
2.8.2. Total Flavonoid Content Measurement
2.8.3. Total Antioxidant Capacity Measurement
2.8.4. Total Reducing Power Measurement
2.9. DPPH Free Radical Scavenging Assay
2.10. Measurement of Anticancerous Activity through MTT Assay
2.11. Statistical Analysis
3. Results
3.1. DNA Barcoding for Identification of Plant
3.2. Transformation and Regeneration
3.3. Molecular Analysis
3.4. HPLC-DAD-Based Quantification of Flavonoids
3.5. Expression Analysis of Flavonoid Biosynthetic Pathway Genes through Real-Time qPCR
3.6. Analyzation of the Antioxidant Potential of Rol Gene Transformed and Untransformed A. carvifolia
3.7. DPPH Free Radical Scavenging Assay
3.8. Measurement of Anticancerous Activity against Cancer Cell Lines through MTT Assay
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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S. No. | Standard Flavonoids | Signal Wavelength (nm) | Retention Time (min) |
---|---|---|---|
1 | Vanillic acid | 257 | 9.116 |
2 | Rutin | 257 | 12.649 |
3 | Gallic acid | 279 | 3.679 |
4 | Catechin | 279 | 7.003 |
5 | syringic acid | 279 | 9.76 |
6 | Coumaric acid | 279 | 13.79 |
7 | Geutisic acid | 325 | 7.433 |
8 | Caffeic acid | 325 | 9.252 |
9 | Ferulic acid | 325 | 12.642 |
10 | Cinnanic acid | 325 | 13.79 |
Source of Variation | Degrees of Freedom | Sum of Squares | Mean Square | F-Value | Prob. |
---|---|---|---|---|---|
Polyphenols | 9 | 117.4 | 13.04 | 54.64 | 0.0000 |
Transgenic lines | 5 | 120.2 | 24.04 | 1592 | 0.0000 |
Interaction | 45 | 20.15 | 0.4478 | 2933 | 0.0000 |
Residual (error) | 120 | 0.9835 | 0.008196 | ||
Total | 179 | 258.7 |
Source of Variation | Degrees of Freedom | Sum of Squares | Mean Square | F-Value | Prob. |
---|---|---|---|---|---|
Interaction | 15 | 3330 | 222.0 | 68.68 | 0.0000 |
Transgenic lines | 5 | 7898 | 1580 | 488.8 | 0.0000 |
Antioxidant assays | 3 | 56270 | 18760 | 5804 | 0.0000 |
Residual (error) | 48 | 155.1 | 3.232 | ||
Total | 71 | 67660 |
Source of Variation | Degrees of Freedom | Sum of Squares | Mean Square | F-Value | Prob. |
---|---|---|---|---|---|
Interaction | 15 | 472.6 | 31.51 | 17.32 | 0.0000 |
Transgenic lines | 5 | 3281 | 656.2 | 360.7 | 0.0000 |
Concentrations | 3 | 22130 | 7376 | 4055 | 0.0000 |
Residual (error) | 48 | 87.33 | 1.819 | ||
Total | 71 | 25970 |
Source of Variation | Degrees of Freedom | Sum of Squares | Mean Square | F-Value | Prob. |
---|---|---|---|---|---|
Cancer cell lines | 2 | 202.2 | 101.1 | 118.2 | 0.0000 |
Transgenic lines | 6 | 34110 | 5684 | 6645 | 0.0000 |
Interaction | 12 | 291.4 | 24.28 | 28.38 | 0.0000 |
Residual (error) | 42 | 35.93 | 0.8554 | ||
Total | 62 | 34640 |
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Khan, A.N.; Dilshad, E. Enhanced Antioxidant and Anticancer Potential of Artemisia carvifolia Buch Transformed with rol A Gene. Metabolites 2023, 13, 351. https://doi.org/10.3390/metabo13030351
Khan AN, Dilshad E. Enhanced Antioxidant and Anticancer Potential of Artemisia carvifolia Buch Transformed with rol A Gene. Metabolites. 2023; 13(3):351. https://doi.org/10.3390/metabo13030351
Chicago/Turabian StyleKhan, Amna Naheed, and Erum Dilshad. 2023. "Enhanced Antioxidant and Anticancer Potential of Artemisia carvifolia Buch Transformed with rol A Gene" Metabolites 13, no. 3: 351. https://doi.org/10.3390/metabo13030351