Alternate identifier:
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Related identifier:
(Is Supplement To) 10.1111/pbi.14261 - DOI
Creator/Author:
Grützner, Ramona [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

König, Kristin [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Horn, Claudia [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Engler, Carola [Icon Genetics GmbH, Weinbergweg 22, 06120 Halle (Saale), Germany]

Laub, Annegret https://orcid.org/0000-0001-6633-3439 [Leibniz Institute of Plant Biochemistry, Dept. of Bioorganic Chemistry, Weinberg 3, 06120 Halle (Saale), Germany]

Vogt, Thomas https://orcid.org/0000-0001-6155-3360 [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]

Marillonnet, Sylvestre https://orcid.org/0000-0002-4263-797X [Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany]
Contributors:
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Title:
Supporting Information: A Transient Expression Tool Box for Anthocyanin Biosynthesis in Nicotiana benthamiana
Additional titles:
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Description:
(Abstract) Transient expression in Nicotiana benthamiana offers a robust platform for the rapid production of complex secondary metabolites. It has proven highly effective in helping identify genes associated with pathways responsible for synthesizing various valuable natural compounds. While this approach has seen considerable success, it has yet to be applied to uncovering genes involved in anthocyanin biosynthetic pathways. This is because only a single anthocyanin, delphinidin 3-O-rutinoside, can be produced in N. benthamiana by activation of anthocyanin biosynthesis using transcription factors. The production of other anthocyanins would necessitate the suppression of certain endogenous flavonoid biosynthesis genes while transiently expressing others. In this work, we present a series of tools for reconstitution of anthocyanin biosynthetic pathways in N. benthamiana leaves. These tools include constructs for expression or silencing of anthocyanin biosynthetic genes and a mutant N. benthamiana line generated using CRISPR. By infiltration of defined sets of constructs, the basic anthocyanins pelargonidin 3-O-glucoside, cyanidin 3-O-glucoside and delphinidin 3-O-glucoside could be obtained in high amount in a few days. Additionally, co-infiltration of supplementary pathway genes enabled the synthesis of more complex anthocyanins. These tools should be useful to identify genes involved in the biosynthesis of complex anthocyanins. They also make it possible to produce novel anthocyanins not found in nature. As an example, we reconstituted the pathway for biosynthesis of Arabidopsis anthocyanin A5, a cyanidin derivative, and achieved the biosynthesis of the pelargonidin and delphinidin variants of A5, pelargonidin A5 and delphinidin A5.
Keywords:
Anthocyanins
plant secondary metabolites
biosynthetic pathway
transient expression
Nicotiana benthamiana
plant biotechnology
synthetic biology
Related information:
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Language:
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Production year:
Subject areas:
Biochemistry
Resource type:
Dataset
Data source:
(Instrument) Waters e2695 chromatography work station
(Instrument) photodiode array detector (PDA)
(Instrument) QDA-mass detector (Waters)
Software used:
Resource production
Software:
Waters Empower - 3
Alternative software:
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Software used:
Resource processing
Software:
Genious Prime Molecular Biology and Bioinformatics - unspecified
Alternative software:
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Software used:
Resource processing
Software:
Microsoft Office - 2013
Alternative software:
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Data processing:
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Publication year:
Rights holders:
Leibniz Institute of Plant Biochemistry, Dept. of Cell and Metabolic Biology, Weinberg 3, 06120 Halle (Saale), Germany
Funding:
-
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Status:
Published
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1670fa88949cc897f41b44d58d08c11a
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Archiving date:
2023-12-06
Archive size:
1.0 GB
Archive creator:
1670fa88949cc897f41b44d58d08c11a
Archive checksum:
a2095d7cac7a3eba2a2fdb6afaef9d76 (MD5)
Embargo period:
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