A Genome-Wide Identification Study Reveals That HmoCYP76AD1, HmoDODAα1 and HmocDOPA5GT Involved in Betalain Biosynthesis in Hylocereus
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Materials
2.2. Gene Identification and Phylogenetic Analyses
2.3. Chromosomal Distribution Analysis
2.4. Relative Expression Analyses by qRT-PCR
2.5. Full-Length cDNA Amplification and Plasmid Construction
2.6. Subcellular Localization Assay
2.7. VIGS of Candidate Genes in H. monacanthus
2.8. Genetic Transformation of A. thaliana
2.9. Transient Expression Assay in N. benthamiana
2.10. Recombinant Expression in Yeast
2.11. Statistical Analysis
3. Results
3.1. Identification and Classification of Candidate Genes Involved in Betalain Biosynthesis
3.2. Genomic Distribution among Pitaya Chromosomes
3.3. Gene Characteristics and Structures
3.4. Expression Patterns of Candidate Genes Involved in Betalain Biosynthesis
3.5. Subcellular Localizations
3.6. VIGS Analyses of Candidate Genes
3.7. Overexpression of HmoDODAα1 and HmoDODAα2 in Arabidopsis and Yeast
3.8. Co-Expression of HmoCYP76AD1, HmoDODAα1/α2 and HmoB5GT1/HmocDOPA5GT in N. benthamiana
3.9. Recombinant Expression of HmoCYP76AD1, HmoDODAα1 and HmocDOPA5GT in Yeast
4. Discussion
4.1. Identification of Genes Involved in Betalain Biosynthesis
4.2. HmoCYP76AD1 Catalyzes Tyrosine to L-DOPA and to Cyclo-DOPA
4.3. HmoDODAα1 Exhibits a High Degree of L-DOPA 4,5-Dioxygenase Activity
4.4. Reconstruction of Heterogenous Production of Betalain Biosynthesis Pathway
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Hua, Q.; Chen, C.; Xie, F.; Zhang, Z.; Zhang, R.; Zhao, J.; Hu, G.; Qin, Y. A Genome-Wide Identification Study Reveals That HmoCYP76AD1, HmoDODAα1 and HmocDOPA5GT Involved in Betalain Biosynthesis in Hylocereus. Genes 2021, 12, 1858. https://doi.org/10.3390/genes12121858
Hua Q, Chen C, Xie F, Zhang Z, Zhang R, Zhao J, Hu G, Qin Y. A Genome-Wide Identification Study Reveals That HmoCYP76AD1, HmoDODAα1 and HmocDOPA5GT Involved in Betalain Biosynthesis in Hylocereus. Genes. 2021; 12(12):1858. https://doi.org/10.3390/genes12121858
Chicago/Turabian StyleHua, Qingzhu, Canbin Chen, Fangfang Xie, Zhike Zhang, Rong Zhang, Jietang Zhao, Guibing Hu, and Yonghua Qin. 2021. "A Genome-Wide Identification Study Reveals That HmoCYP76AD1, HmoDODAα1 and HmocDOPA5GT Involved in Betalain Biosynthesis in Hylocereus" Genes 12, no. 12: 1858. https://doi.org/10.3390/genes12121858