Leaves of Moringa oleifera Are Potential Source of Bioactive Compound β-Carotene: Evidence from In Silico and Quantitative Gene Expression Analysis
Abstract
:1. Introduction
2. Results
2.1. Retrieval of Zeta-Carotene Desaturase (ZDS) and Phytoene Synthase (PSY) Genes
2.2. Sequencing of ZDS and PSY Genes of M. oleifera and Submission to NCBI Database
2.3. Multiple Sequence Alignments and Phylogenetic Analysis
2.4. RNA Extraction and cDNA Preparation
2.5. qRT-PCR Data Interpretation
3. Discussion
4. Materials and Methods
4.1. Assembling the Genes Data Set
4.2. DNA Extraction, PCR and Sequencing of ZDS and PSY Genes
4.3. Multiple Sequence Alignment and Phylogenetic Analysis
4.4. Protein Profiling and Analysis
4.5. Primer Design
4.6. Sample Collection and RNA Extraction
4.7. cDNA Preparation
4.8. qRT-PCR Analysis
4.9. Quantitative Gene Expression Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Species Name | Gene Name | E-Value | Identity (%) | Domain | Family |
---|---|---|---|---|---|
Arabidopsis thaliana | ZDS | 0.0 | 98.0 | PLN02487 | Amino oxidase |
PSY | 0.0 | 97.5 | PLN02632 | Squalene Synthase (SQS) | |
Brassica napus | ZDS | 1 × 10−121 | 83.4 | PLN02487 | Amino oxidase |
PSY | 1 × 10−140 | 83.5 | PLN02632 | Squalene Synthase (SQS) | |
Brassica rapa | ZDS | 1 × 10−11 | 76.4 | PLN02487 | Amino oxidase |
PSY | 1 × 10−148 | 82.3 | PLN02632 | Squalene Synthase (SQS) | |
Brassica oleracea | ZDS | 1 × 10−13 | 86.0 | PLN02487 | Amino oxidase |
PSY | 1 × 10−148 | 83.5 | PLN02632 | Squalene Synthase (SQS) |
Genes | Expression Changes Relative to That of A. thaliana | ||||
---|---|---|---|---|---|
Arabidopsis thaliana | Brassica napus | Brassica rapa | Bixa orellana | Moringa oleifera | |
ZDS | ↑651.3 * | ↑412.8 * | ↑615.0 * | ↓140.8 * | ↓140.8 * |
PSY | ↓412.3 * | ↑536.1 * | ↓341.8 | ↓514.1 * | ↓514.1 * |
Actin | ↑120.9 * | ↑102.6 * | ↑7.4 * | ↓27.0 * | ↓17.0 * |
S. No. | Genes | Primers | Length | TM (°C) | GC (%) |
---|---|---|---|---|---|
1. | PSY | Forward. TGCAGCTTAAACGAGCAAGA | 20 | 59.90 | 45 |
Reverse. AGCAATGAAGCCCATACCTG | 20 | 60.10 | 50 | ||
2. | ZDS | Forward. GACTCCGATGTTTCCGACAT | 20 | 59.93 | 50 |
Reverse. CACTTTGCCACCAATGAATG | 20 | 59.96 | 45 |
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Muteeb, G.; Aatif, M.; Farhan, M.; Alsultan, A.; Alshoaibi, A.; Alam, M.W. Leaves of Moringa oleifera Are Potential Source of Bioactive Compound β-Carotene: Evidence from In Silico and Quantitative Gene Expression Analysis. Molecules 2023, 28, 1578. https://doi.org/10.3390/molecules28041578
Muteeb G, Aatif M, Farhan M, Alsultan A, Alshoaibi A, Alam MW. Leaves of Moringa oleifera Are Potential Source of Bioactive Compound β-Carotene: Evidence from In Silico and Quantitative Gene Expression Analysis. Molecules. 2023; 28(4):1578. https://doi.org/10.3390/molecules28041578
Chicago/Turabian StyleMuteeb, Ghazala, Mohammad Aatif, Mohd Farhan, Abdulrahman Alsultan, Adil Alshoaibi, and Mir Waqas Alam. 2023. "Leaves of Moringa oleifera Are Potential Source of Bioactive Compound β-Carotene: Evidence from In Silico and Quantitative Gene Expression Analysis" Molecules 28, no. 4: 1578. https://doi.org/10.3390/molecules28041578