Cyanidin based anthocyanin biosynthesis in orange carrot is restored by expression of AmRosea1 and AmDelila, MYB and bHLH transcription factors

Sharma, Shrikant; Holme, Inger B.; Dionisio, Giuseppe; Kodama, Miyako; Dzhanfezova, Tsaneta; Joernsgaard, Bjarne; Brinch-Pedersen, Henrik

doi:10.1007/s11103-020-01002-1

Published: 08 April 2020

Cyanidin based anthocyanin biosynthesis in orange carrot is restored by expression of AmRosea1 and AmDelila, MYB and bHLH transcription factors

Plant Molecular Biology volume 103, pages 443–456 (2020)Cite this article

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Abstract

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The simultaneous expression of AmRosea1 and AmDelila transcription factors from snapdragon can activate the anthocyanin pathway in orange carrots, leading to the synthesis and accumulation of anthocyanins in the taproots.

Abstract

Anthocyanins are phenolic compounds produced in various parts of plants. They are used as natural food dyes and are reported as beneficial antioxidants for humans. Black carrot is an important source for anthocyanins; however, the reason for the lack of anthocyanin production in the orange carrot is unknown. Anthocyanins are synthesized by a specific branch of the phenylpropanoid pathway that has previously been reported to be activated by a triad of R2R3-MYB, basic helix-loop helix (bHLH) and WD40 transcription factors (TFs). In the current study, orange carrots were turned purple by simultaneous expression of R2R3-MYB and bHLH TFs, i.e. AmRosea1 and AmDelila from snapdragon (Antirrhinum majus). Simultaneous transgenic expression of the TFs under a constitutive promoter in the orange carrot cultivar ‘Danvers 126’ lead to consistent upregulation of anthocyanin-related biosynthetic genes and significant accumulation of anthocyanins in leaves, stems and taproots. Highest overall content of soluble anthocyanins in the taproot among the transformants amounted to 44.38 mg g⁻¹ dry weight. The anthocyanin profile of the transformants were significantly different from the profile in the reference black carrot ‘Deep Purple’. The main anthocyanins present in the transformed taproots were cyanidin 3-xylosyl(sinapoylglucosyl)galactoside, whereas the main anthocyanin present in Deep Purple was cyanidin 3-xylosyl(feruloylglucosyl)galactoside. This study confirms the presence of the necessary biosynthetic genes in orange carrots for production of anthocyanins and demonstrates the absence of suitable R2R3-MYB and bHLH TFs for stimulating anthocyanin biosynthesis in the orange carrot.

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Funding

Funding was provided by Innovationsfonden (Grant No. 4105-00006B).

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Department of Molecular Biology and Genetics, Aarhus University, Flakkebjerg, 4200, Slagelse, Denmark
Shrikant Sharma, Inger B. Holme, Giuseppe Dionisio & Henrik Brinch-Pedersen
Centre for GeoGenetics, Section for Evolutionary Genomics, Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark
Miyako Kodama
Chr. Hansen Natural Colors A/S, Agern Alle 24, 2970, Hoersholm, Denmark
Tsaneta Dzhanfezova & Bjarne Joernsgaard

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Shrikant Sharma
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Inger B. Holme
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Giuseppe Dionisio
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Miyako Kodama
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Tsaneta Dzhanfezova
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Bjarne Joernsgaard
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Henrik Brinch-Pedersen
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Correspondence to Henrik Brinch-Pedersen.

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Sharma, S., Holme, I.B., Dionisio, G. et al. Cyanidin based anthocyanin biosynthesis in orange carrot is restored by expression of AmRosea1 and AmDelila, MYB and bHLH transcription factors. Plant Mol Biol 103, 443–456 (2020). https://doi.org/10.1007/s11103-020-01002-1

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Received: 13 September 2019
Accepted: 25 March 2020
Published: 08 April 2020
Issue Date: July 2020
DOI: https://doi.org/10.1007/s11103-020-01002-1

Keywords

AmRosea1
AmDelila
Anthocyanins
Daucus carota L.
Orange carrots
Black carrots
Transcription factors

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Funding

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Corresponding author

Additional information

Publisher's Note

Electronic supplementary material

Supplementary file1 (XLSX 13 kb)

Supplementary file2 (DOCX 40 kb)

Supplementary file3 (DOCX 31 kb)

Supplementary file4 (DOCX 18 kb)

Supplementary file5 (DOCX 58 kb)

Supplementary file6 (DOCX 118 kb)

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