Jasmonates are plant hormones that induce the accumulation of many secondary metabolites, such as rutin in buckwheat, via regulation of jasmonate-responsive transcription factors. Here, we report on the identification of a clade of jasmonate-responsive subgroup 4 MYB transcription factors, FtMYB13, FtMYB14, FtMYB15, and FtMYB16, which directly repress rutin biosynthesis in Fagopyrum tataricum. Immunoblot analysis showed that FtMYB13, FtMYB14, and FtMYB15 could be degraded via the 26S proteasome in the COI1-dependent jasmonate signaling pathway, and that this degradation is due to the SID motif in their C-terminus. Yeast two-hybrid and bimolecular fluorescence complementation assays revealed that FtMYB13, FtMYB14, and FtMYB15 interact with the importin protein Sensitive to ABA and Drought 2 (FtSAD2) in stem and inflorescence. Furthermore, the key repressor of jasmonate signaling FtJAZ1 specifically interacts with FtMYB13. Point mutation analysis showed that the conserved Asp residue of the SID domain contributes to mediating protein-protein interaction. Protoplast transient activation assays demonstrated that FtMYB13, FtMYB14, and FtMYB15 directly repress phenylalanine ammonia lyase (FtPAL) gene expression, and FtSAD2 and FtJAZ1 significantly promote the repressing activity of FtMYBs. These findings may ultimately be promising for further engineering of plant secondary metabolism.

Center of Plant Systems Biology and Biotechnology Plovdiv Bulgaria

College of Agricultural Science Xichang University Xichang Sichuan China

College of Agriculture Hainan University Haikou Hainan China

College of Agriculture Inner Mongolia Agricultural University Hohhot Inner Mongolia China

College of Landscape and Travel Agricultural University of Hebei Baoding China

Department of Evolutionary Biochemistry AN Belozersky Institute of Physico Chemical Biology MV Lomonosov Moscow State University Moscow Russia

Department of Gene Bank Crop Research Institute Drnovská Czech Republic

Grassland Institute China Agricultural University Beijing China

Group of Plant Cell Biotechnology and Metabolomics The Stephan Angeloff Institute of Microbiology Bulgarian Academy of Sciences Plovdiv Bulgaria

Institute of Crop Science Chinese Academy of Agricultural Sciences Beijing China

School of Life Sciences Hunan University of Science and Technology Xiangtan Hunan China

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Fagopyrum esculentum ssp. ancestrale-A Hybrid Species Between Diploid F. cymosum and F. esculentum