Three bilberry bHLHs of subgroup IIIf play divergent roles in fruit anthocyanin and flavonoid biosynthesis.

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Main Authors: Karppinen, Katja, Raami, Lauri, Edema, Hilary, Ashraf, Muhammad Furqan, Salo, Heikki M, Espley, Richard V, Jaakola, Laura
Format: Artículo científico
Language:en
Published: Scientific reports 2025
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author Karppinen, Katja
Raami, Lauri
Edema, Hilary
Ashraf, Muhammad Furqan
Salo, Heikki M
Espley, Richard V
Jaakola, Laura
author_facet Karppinen, Katja
Raami, Lauri
Edema, Hilary
Ashraf, Muhammad Furqan
Salo, Heikki M
Espley, Richard V
Jaakola, Laura
Karppinen, Katja
Raami, Lauri
Edema, Hilary
Ashraf, Muhammad Furqan
Salo, Heikki M
Espley, Richard V
Jaakola, Laura
collection PubMed - marine biology
contents Three bilberry bHLHs of subgroup IIIf play divergent roles in fruit anthocyanin and flavonoid biosynthesis. Karppinen, Katja Raami, Lauri Edema, Hilary Ashraf, Muhammad Furqan Salo, Heikki M Espley, Richard V Jaakola, Laura Anthocyanins Fruit Gene Expression Regulation, Plant Flavonoids Basic Helix-Loop-Helix Proteins Phylogeny Vaccinium myrtillus Plant Proteins Nicotiana Bilberry (Vaccinium myrtillus L.) fruit are one of the best natural sources of anthocyanins. Anthocyanin and flavonoid biosynthesis are transcriptionally regulated by the conserved MBW complex, including R2R3 MYB, basic helix-loop-helix (bHLH) and WD40 proteins. In this study, homology-search in the bilberry genome identified three putative anthocyanin biosynthesis-regulating bHLH genes, all containing a MYB binding domain, indicating their possible interaction with R2R3 MYBs. Phylogenetic analysis assigned the VmbHLHs to three different clusters within the IIIf subgroup of bHLHs. The expression of VmbHLH2 elevated at berry ripening, while VmbHLH1 and VmbHLH3 showed decreasing expression towards berry maturation. When transiently overexpressed together with MdMYB10 in Nicotiana benthamiana leaves, VmbHLH2 and VmbHLH3 were able to complement MdbHLH3 and upregulate a full set of anthocyanin biosynthetic genes leading to anthocyanin accumulation. In contrast, VmbHLH1 induced a reduced set of flavonoid pathway genes. Our results provide the first functional characterization of flavonoid-regulating bHLHs that contribute to the MBW complex in the economically important genus Vaccinium. We propose that the TT8-type VmbHLH2 is involved in anthocyanin biosynthesis in ripening bilberry fruit, while the MYC1-type VmbHLH1 and GL3/EGL3-type VmbHLH3 are associated with anthocyanin and PA biosynthesis, mainly in tissues other than berries.
format Artículo científico
id pubmed_40813431
institution PubMed
language en
publishDate 2025
publisher Scientific reports
record_format pubmed
spellingShingle Three bilberry bHLHs of subgroup IIIf play divergent roles in fruit anthocyanin and flavonoid biosynthesis.
Karppinen, Katja
Raami, Lauri
Edema, Hilary
Ashraf, Muhammad Furqan
Salo, Heikki M
Espley, Richard V
Jaakola, Laura
Anthocyanins
Fruit
Gene Expression Regulation, Plant
Flavonoids
Basic Helix-Loop-Helix Proteins
Phylogeny
Vaccinium myrtillus
Plant Proteins
Nicotiana
Three bilberry bHLHs of subgroup IIIf play divergent roles in fruit anthocyanin and flavonoid biosynthesis. Karppinen, Katja Raami, Lauri Edema, Hilary Ashraf, Muhammad Furqan Salo, Heikki M Espley, Richard V Jaakola, Laura Anthocyanins Fruit Gene Expression Regulation, Plant Flavonoids Basic Helix-Loop-Helix Proteins Phylogeny Vaccinium myrtillus Plant Proteins Nicotiana Bilberry (Vaccinium myrtillus L.) fruit are one of the best natural sources of anthocyanins. Anthocyanin and flavonoid biosynthesis are transcriptionally regulated by the conserved MBW complex, including R2R3 MYB, basic helix-loop-helix (bHLH) and WD40 proteins. In this study, homology-search in the bilberry genome identified three putative anthocyanin biosynthesis-regulating bHLH genes, all containing a MYB binding domain, indicating their possible interaction with R2R3 MYBs. Phylogenetic analysis assigned the VmbHLHs to three different clusters within the IIIf subgroup of bHLHs. The expression of VmbHLH2 elevated at berry ripening, while VmbHLH1 and VmbHLH3 showed decreasing expression towards berry maturation. When transiently overexpressed together with MdMYB10 in Nicotiana benthamiana leaves, VmbHLH2 and VmbHLH3 were able to complement MdbHLH3 and upregulate a full set of anthocyanin biosynthetic genes leading to anthocyanin accumulation. In contrast, VmbHLH1 induced a reduced set of flavonoid pathway genes. Our results provide the first functional characterization of flavonoid-regulating bHLHs that contribute to the MBW complex in the economically important genus Vaccinium. We propose that the TT8-type VmbHLH2 is involved in anthocyanin biosynthesis in ripening bilberry fruit, while the MYC1-type VmbHLH1 and GL3/EGL3-type VmbHLH3 are associated with anthocyanin and PA biosynthesis, mainly in tissues other than berries.
title Three bilberry bHLHs of subgroup IIIf play divergent roles in fruit anthocyanin and flavonoid biosynthesis.
topic Anthocyanins
Fruit
Gene Expression Regulation, Plant
Flavonoids
Basic Helix-Loop-Helix Proteins
Phylogeny
Vaccinium myrtillus
Plant Proteins
Nicotiana
url https://pubmed.ncbi.nlm.nih.gov/40813431/