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FtMYB18 acts as a negative regulator of anthocyanin/proanthocyanidin biosynthesis in Tartary buckwheat

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Abstract

Key message

FtMYB18 plays a role in the repression of anthocyanins and proanthocyanidins accumulation by strongly down-regulating the CHS and DFR genes in Tartary buckwheat, and the C5 motif plays an important role in this process.

Abstract

Anthocyanins and proanthocyanidins (PAs) are important flavonoids in Tartary buckwheat (Fagopyrum tataricum Gaertn.), which provides various vibrant color and stronge abiotic stress resistance. Their synthesis is generally regulated by MYB transcription factors at transcription level. However, the negative regulations of MYB and their effects on flavonol metabolism are poorly understood. A SG4-like MYB subfamily TF, FtMYB18, containing C5 motif was identified from Tartary buckwheat. The expression of FtMYB18 was not only showed a negative correlation with anthocyanins and PAs content but also strongly respond to MeJA and ABA. As far as the transgenic lines with FtMYB18 overexpression, anthocyanins and PAs accumulations were decreased through down-regulating expression levels of NtCHS and NtDFR in tobacco, AtDFR and AtTT12 in Arabidopsis, FtCHS, FtDFR and FtANS in Tartary buckwheat hairy roots, respectively. However, FtMYB18 showed no effect on the FLS gene expression and the metabolites content in flavonol synthesis branch. The further molecular interaction analysis indicated FtMYB18 could mediate the inhibition of anthocyanins and PAs synthesis by forming MBW transcriptional complex with FtTT8 and FtTTG1, or MYB-JAZ complex with FtJAZ1/-3/-4/-7. Importantly, in FtMYB18 mutant lines with C5 motif deletion (FtMYB18-C), both of anthocyanins and PAs accumulations had recovered to the similar level as that in wild type, which was attributed to the weakened MBW complex activity or the deficient molecular interaction between FtMYB18ΔC5 with FtJAZ3/-4. The results showed that FtMYB18 could suppress anthocyanins and PAs synthesis at transcription level through the specific interaction of C5 motif with other proteins in Tartary buckwheat.

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Abbreviations

ABA:

Abscisic acid

ANS:

Anthocyanidin synthase

CHI:

Chalcone isomerase

CHS:

Chalcone synthase

DFR:

Dihydroflavonol reductase

EBGs:

Early biosynthetic genes

F3′H:

Flavonoid 3′-hydroxylase

FLS:

Flavonol synthase

LBGs:

Late biosynthetic genes

MBW:

MYB-bHLH-WD40

MeJA:

Methyl jasmonate

PAs:

Proanthocyanidins

SG4:

Sub-group 4

SG5:

Sub-group 5

SG6:

Sub-group 6

SG7:

Sub-group 7

SPSS:

Statistical product and service solutions

TFs:

Transcription factors

TT12:

Transparent testa 12

UFGT:

UDP-flavonoid glucosyltransferase

X-gal:

5-Bromo-4-chloro-3indolyl β-d-galactopyranoside

X-α-Gal:

5-Bromo-4-chloro-3-indoxyl a-Dgalactoside

JAZ:

Jasmonate ZIM-domain

HCl:

Hydrochloric acid

WT:

Wild type

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Acknowledgements

We thank the Annick Bleys for critical reading and editing of this manuscript.

Funding

This work was supported by the National Natural Science Foundation of China (Project No. 31871698).

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QXD and HXZ conceived the original screening and research plans; YJH, YC, MW and ZXZ carried out part of material collection, RNA extraction; CLL carried out flavonoid quantification analysis; XLW and HC performed most of the experiments; PFY analyzed the data, QXD and QW design most of the experiments and wrote the article; All authors read and approved the final manuscript.

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Correspondence to Qi Wu.

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Dong, Q., Zhao, H., Huang, Y. et al. FtMYB18 acts as a negative regulator of anthocyanin/proanthocyanidin biosynthesis in Tartary buckwheat. Plant Mol Biol 104, 309–325 (2020). https://doi.org/10.1007/s11103-020-01044-5

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