Gene Cloning,Subcellular Localization and Expression Analyses of FtMYBF Transcription Factor in Fagopyrum tataricum

doi:10.13560/j.cnki.biotech.bull.1985.2020-0861

Abstract

Abstract:

The SG7 subfamily members of R2R3-MYB transcription factors play a critical role in the biosynthesis of flavonols in plants. Exploring the function of SG7 R2R3-MYB transcription factors in flavonol biosynthesis in Fagopyrum tataricum would lay a foundation for the molecular regulation mechanism of flavonol biosynthesis in F. tataricum. RT-PCR was used to clone 1 SG7 R2R3-MYB transcription factor identified by the combined analysis of transcriptome and metabolome in previous study,and named as FtMYBF. This gene then was analyzed by bioinformatics,subcellular localization,gene coexpression and the correlation between gene expression level and total flavonoid content. As a result,the CDS length of FtMYBF was 1 119 bp and encoded 372 amino acids. Multiple sequence alignment and phylogenetic tree analyses showed that FtMYBF belonged to R2R3-MYB SG7 subfamily. Subcellular localization results suggested that the FtMYBF protein was located in the nucleus. Gene coexpression analysis demonstrated that FtMYBF was coexpressed with several structural genes of flavonol biosynthesis in F. tataricum. The correlation analysis between gene expression and total flavonoids content indicated that there was a highly positive correlation between the FtMYBF expression level and the total flavonoid content in different tissues. The results of this study suggest that FtMYBF is a member of R2R3-MYB SG7 subfamily and is a nuclear localization transcription factor,which might positively regulate the flavonols biosynthesis by regulating the expression of the flavonol biosynthesis structural genes in F. tataricum.

Key words: Fagopyrum tataricum, FtMYBF, R2R3-MYB transcription factor, gene cloning, subcellular localization, flavonoid

SUN Xiao-qian, WANG Jia-rui, CHEN Qing-fu, LI Hong-you. Gene Cloning,Subcellular Localization and Expression Analyses of FtMYBF Transcription Factor in Fagopyrum tataricum[J]. Biotechnology Bulletin, 2021, 37(3): 10-17.

Figures/Tables 7

References 32

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引物名称	引物序列（5'-3'）
FtMYBF-f1	ATGGGAAGAGCACCTTGCTG
FtMYBF-r1	CTCAAGAAAGAAGCCAGGCAAC
FtMYBF-f2	CAAGCTTGCATGCCTGCAGGTCGACATGGGAA- GAGCACCTTGCTG
FtMYBF-r2	CCTCGCCCTTGCTCACCATGGATCCAGAAAGA- AGCCAGGCAACCA
FtMYBF-qf	GGGAGAGAGTAGCTGCAATG
FtMYBF-qr	CCCACATTAGCCACGATAGG
FtActin7-f	ATGTTCACTACCACCGCTGA
FtActin7-r	TGAACCTCTCAGCACCAATC

引物名称	引物序列（5'-3'）
FtMYBF-f1	ATGGGAAGAGCACCTTGCTG
FtMYBF-r1	CTCAAGAAAGAAGCCAGGCAAC
FtMYBF-f2	CAAGCTTGCATGCCTGCAGGTCGACATGGGAA- GAGCACCTTGCTG
FtMYBF-r2	CCTCGCCCTTGCTCACCATGGATCCAGAAAGA- AGCCAGGCAACCA
FtMYBF-qf	GGGAGAGAGTAGCTGCAATG
FtMYBF-qr	CCCACATTAGCCACGATAGG
FtActin7-f	ATGTTCACTACCACCGCTGA
FtActin7-r	TGAACCTCTCAGCACCAATC