Schilbert, Hanna; Schöne, Maximilian; Baier, Thomas ; Busche, Mareike ; Viehöver, Prisca; Weisshaar, Bernd ; Holtgräwe, Daniela: Characterization of the Brassica napus flavonol synthase gene family reveals bifunctional flavonol synthases. In: Frontiers in Plant Science. Jg.12. 2021
Inhalt
- Plant Material
- Identification of BnaFLS Candidate Genes
- Sequence-Specific Analyses of BnaFLS Candidates and Secondary Structure Modeling
- Gene Expression Analysis: Ribonucleic Acid Extraction, Library Construction, and Sequencing
- Gene Expression Analysis and Co-expression Analysis Using Brassica napus RNA-Seq Data
- Functional Annotation of Brassica napus Express 617 Genes
- Generation of BnaFLSs Constructs
- Flavanone 3-Hydroxylase and Flavonol Synthase Bioconversion Assay in E. coli
- Generation of Complementation Lines
- Flavonol Content Analysis by High-Performance Thin-Layer Chromatography
- In situ Flavonoid Staining of Whole Seedlings
- Results
- Flavonol Synthase Family of Brassica napus
- Organ- and Temporal-Specific Expression of BnaFLS Candidates
- BnaFLS1-1 and BnaFLS1-2 Are Co-expressed With Major Players of the Flavonoid Biosynthesis
- BnaFLS Candidates Share High Amino Acid Sequence Identity to Arabidopsis thaliana 2-ODD Orthologs
- BnaFLS Candidates Carry Residues Important for Flavonol Synthase and Flavanone 3-Hydroxylase Activity
- Functional Characterization of BnaFLS Candidates
- BnaFLS1-1 and BnaFLS1-2 Are Bifunctional Enzymes Exhibiting Flavanone 3-Hydroxylase and Flavonol Synthase Activity
- BnaFLS Family Members With Divergent Enzyme Functionalities
- Structural Modeling Revealed Three Major Differences of the Bifunctional Enzymes Compared to Monofunctional Ones
- Discussion
- Phylogeny of BnaFLS Gene Family Members
- The BnaFLS Gene Family Contains Two Bifunctional Flavonol Synthases
- BnaFLS1s Are Major Players in Flavonol Biosynthesis in Brassica napus Seeds
- Future Perspectives in Engineering Flavonol Content in Brassica napus
- Data Availability Statement
- Author Contributions
- Funding
- Acknowledgments
- Supplementary Material
- References