Production of Genistein in Amaranthus tricolor var. tristis and Spinacia oleracea by Expression of Glycine max Isoflavone Synthase
Abstract
:1. Introduction
2. Results
2.1. Confirmation of Cloned IFS in Plant Expression Vector
2.2. Transformation of Agrobacterium tumefaciens with GmIFS and Plant Transformation
2.3. HPLC Detection of Naringenin and Genistein in Infiltrated Samples
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Construction of Plant Expression Vector
4.3. Agroinfiltration
4.4. RNA Isolation and cDNA Synthesis
4.5. HPLC Detection
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
CaMV | Cauliflower mosaic virus |
cDNA | Complementary deoxyribonucleic acid |
DNA | Deoxyribonucleic acid |
dNTP’s | Deoxynucleotide triphosphates |
°C | Degrees Celsius |
FW | Fresh weight |
HPLC | High-performance liquid chromatography |
mg L−1 | Milligram per liter |
mL | Milliliter |
nm | Nanometer |
RNA | Ribonucleic acid |
rpm | Revolutions per minute |
µL | Microliter |
µM | Micromolar |
µ | Micron |
U | Unit |
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Reaction Mix | Program | ||
---|---|---|---|
Components | Volume (µL) | 1. Initial Denaturation | 94 °C for 5 min |
Emerald AmpR PCR master mix | 10 | 2. Denaturation | 94 °C for 30 s |
Forward primer (10 pM) | 1 | 3. Annealing | 56 °C for 45 s |
Reverse primer (10 pM) | 1 | 4. Extension | 72 °C for 45 s |
Nuclease-free water | 6 | Step 2–4 repeated 25 cycles | |
DNA template | 2 | Final Extension | 72 °C for 10 min |
Total | 20 | Final Hold | 4 °C |
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Malla, A.; Shanmugaraj, B.; Sharma, A.; Ramalingam, S. Production of Genistein in Amaranthus tricolor var. tristis and Spinacia oleracea by Expression of Glycine max Isoflavone Synthase. Plants 2021, 10, 2311. https://doi.org/10.3390/plants10112311
Malla A, Shanmugaraj B, Sharma A, Ramalingam S. Production of Genistein in Amaranthus tricolor var. tristis and Spinacia oleracea by Expression of Glycine max Isoflavone Synthase. Plants. 2021; 10(11):2311. https://doi.org/10.3390/plants10112311
Chicago/Turabian StyleMalla, Ashwini, Balamurugan Shanmugaraj, Ashutosh Sharma, and Sathishkumar Ramalingam. 2021. "Production of Genistein in Amaranthus tricolor var. tristis and Spinacia oleracea by Expression of Glycine max Isoflavone Synthase" Plants 10, no. 11: 2311. https://doi.org/10.3390/plants10112311