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The RcMYB114 Gene Coordinated Regulation the Anthocyanin Accumulation by Interacting with RcMIEL1 Gene

agricultural-science
plant-science

Abstract

MIEL1 is a RING-type E3 ubiquitin ligase characterize d by a conserved RING domain that catalyzes the ubiquitination of substrate proteins. However, direct experimental evidence on the interaction between E3 ubiquitin ligases and MYB transcription factors in roses is still scarce. In this study, conversion experiments in tobacco leaves and rose petals revealed that the RcMYB114 gene functions as a regulatory gene, promoting anthocyanin accumulation in both tissues and determining their color. Utilizing RcMYB114 as a bait gene, we identified the RcMIEL1 gene. The RcMIEL1 gene was cloned and mapped to chromosome 4, consisting of 3099 base pairs (bp), including 11 introns and 12 exons. This gene has an 804 bp open reading frame and encodes 267 amino acids. Phylogenetic analysis revealed close genetic relationships between RcMIEL1 and that of strawberry. RcMIEL1 was expressed in the young leaves, stems, styles, pollen, and petals of roses, with the highest expression level observed in pollen, the lowest in stems, and a peak during the S4 phase in rose petals. Yeast two-hybrid analysis demonstrated that RcMIEL1 and RcMYB114 proteins interact. Overexpression and gene silencing experiments indicated that RcMIEL 1 alters its expression levels by interacting with RcMYB114. The expression of RcMIEL1 was inversely related to anthocyanin content, whereas the expressions of RcMYB114 and NtUFGT were positively correlated with anthocyanin levels. These findings suggest that the RcMYB114 gene coordinates the regulation of anthocyanin accumulation by interacting with the RcMIEL1 gene. Our findings provide a theoretical foundation for elucidating the molecular mechanisms underlying MYB transcription factor regulation in flower color formation.
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