Dataset Information

Isolation and molecular characterization of a Lotus japonicus R2R3-MYB subgroup 7 transcription factor gene.

ABSTRACT: We isolated an ortholog (LjMYB12) of the Arabidopsis R2R3-MYB transcription factor (TF) gene from Lotus japonicus to investigate the regulation of flavonoid biosynthesis, which is driven by many paralogous genes in L. japonicus. We characterized the spatial and temporal expression of LjMYB12 in leaves, stems, roots, flowers, immature seeds, seedling leaves, and seedling roots. Expression was much higher in flowers than in other tissues. To verify the relationship between the expression of LjMYB12 and that of flavonoid biosynthesis genes, we generated transgenic L. japonicus plants overexpressing LjMYB12. Overexpression of LjMYB12 resulted in the upregulation of genes for a chalcone synthase paralog (CHS1), flavanone 3-hydroxylase, and flavonol synthase. Interestingly, LjMYB12 strongly activated CHS1 but did not activate other CHS paralogs. This result suggests differences in the spatial or temporal activation of CHS paralogs by R2R3-MYB TFs. Molecular characterization of R2R3-MYB TFs in L. japonicus will reveal the effects of gene duplication on the regulation of diverse flavonoid biosynthesis.

SUBMITTER: Kunihiro S

PROVIDER: S-EPMC6543700 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Isolation and molecular characterization of a Lotus japonicus R2R3-MYB subgroup 7 transcription factor gene.

Kunihiro Satoru S Tanabe Daigo D Niwa Yuiko Y Kitamura Keisuke K Abe Jun J Yamada Tetsuya T

Plant biotechnology (Tokyo, Japan) 20170211 1

We isolated an ortholog (LjMYB12) of the Arabidopsis R2R3-MYB transcription factor (TF) gene from Lotus japonicus to investigate the regulation of flavonoid biosynthesis, which is driven by many paralogous genes in L. japonicus. We characterized the spatial and temporal expression of LjMYB12 in leaves, stems, roots, flowers, immature seeds, seedling leaves, and seedling roots. Expression was much higher in flowers than in other tissues. To verify the relationship betw ...[more]

PMID: 31275007

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Project description:Epimedium species have been widely used both as traditional Chinese medicinal plants and ornamental perennials. Both flavonols, acting as the major bioactive components (BCs) and anthocyanins, predominantly contributing to the color diversity of Epimedium flowers belong to different classes of flavonoids. It is well-acknowledged that flavonoid biosynthetic pathway is predominantly regulated by R2R3-MYB transcription factor (TF) as well as bHLH TF and WD40 protein at the transcriptional level. MYB TFs specifically regulating anthocyanin or flavonol biosynthetic pathway have been already isolated and functionally characterized from Epimedium sagittatum, but a R2R3-MYB TF involved in regulating both these two pathways has not been functionally characterized to date in Epimedium plants. In this study, we report the functional characterization of EsMYB9, a R2R3-MYB TF previously isolated from E. sagittatum. The previous study indicated that EsMYB9 belongs to a small subfamily of R2R3-MYB TFs containing grape VvMYB5a and VvMYB5b TFs, which regulate flavonoid biosynthetic pathway. The present studies show that overexpression of EsMYB9 in tobacco leads to increased transcript levels of flavonoid pathway genes and increased contents of anthocyanins and flavonols. Yeast two-hybrid assay indicates that the C-terminal region of EsMYB9 contributes to the autoactivation activity, and EsMYB9 interacts with EsTT8 or AtTT8 bHLH regulator. Transient reporter assay shows that EsMYB9 slightly activates the expression of EsCHS (chalcone synthase) promoter in transiently transformed leaves of Nicotiana benthamiana, but the addition of AtTT8 or EsTT8 bHLH regulator strongly enhances the transcriptional activation of EsMYB9 against five promoters of the flavonoid pathway genes except EsFLS (flavonol synthase). In addition, co-transformation of EsMYB9 and EsTT8 in transiently transfected tobacco leaves strongly induces the expressions of flavonoid biosynthetic genes. The potential role of EsMYB9 in modulating the biosynthesis and accumulation of sucrose-induced anthocyanin and flavonol-derived BCs is also discussed. These findings suggest that EsMYB9 is a novel R2R3-MYB TF, which regulates the flavonoid biosynthetic pathway in Epimedium, but distinctly different with the anthocyanin or flavonol-specific MYB regulators identified previously in Epimedium plants.

Dataset Information

Isolation and molecular characterization of a Lotus japonicus R2R3-MYB subgroup 7 transcription factor gene.

Publications

Isolation and molecular characterization of a <i>Lotus japonicus</i> <i>R2R3-MYB</i> subgroup 7 transcription factor gene.

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