Abstract

Paeonia qiui is a wild tree peony native to China. Its leaves show a clear purple-red color from the germination to the flowering stage, and it has high leaf-viewing value. A MYB transcription factor gene, designated as PqMYB4, was isolated from leaves of P. qiui based on transcriptome datas. The full-length cDNA of PqMYB4 was 693 bp, encoding 230 amino acids. Sequence alignment and phylogenetic analysis revealed that PqMYB4 was a R2R3-MYB transcription factor clustered with AtMYB4 in Arabidopsis thaliana. Moreover, it contained a C1 motif, an EAR repression motif and a TLLLFR motif in the C-terminal domains, which were unique in transcription repression MYB. Subcellular location analysis showed that PqMYB4 was located in the cell nucleus. PqMYB4 was highly expressed in the late stage of leaf development, and was negatively correlated with the anthocyanin content. The petiole of wild-type Arabidopsis seedlings was deeper in color than the transgenic lines of PqMYB4 and showed a little purple-red color. The seed coat color of Arabidopsis seeds that overexpressed PqMYB4 gene was significantly lighter than that of wild-type seeds. In transgenic Arabidopsis, the expression level of AtCHS, AtCHI, AtDFR and AtANS were down-regulated significantly. These results showed that PqMYB4 was involved in the negative regulation of anthocyanin biosynthesis in tree peony leaves, which can control the anthocyanin pathway genes. Together, these findings provide a valuable resource with which to further study the regulatory mechanism of anthocyanin biosynthesis in the leaf of P. qiui. They also benefit the molecular breeding of tree peony cultivars with colored leaf.

Highlights

  • Anthocyanins are a class of flavonoids derived from phenylalanine

  • chalcone synthase (CHS) is the key enzyme in the early stage of anthocyanin biosynthesis, and DFR and anthocyanidin synthase (ANS) are key enzymes in the late stage of anthocyanin synthesis, both of which play an important role in the accumulation of anthocyanins [3,4]

  • This study provided a basis for revealing the mechanism of leaf color change in P. qiui, and provided genetic resources for the molecular breeding of the tree peony cultivars with colored leaf

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Summary

Introduction

Anthocyanins are a class of flavonoids derived from phenylalanine. They are water-soluble pigment that can make plants exhibit colors ranging from orange-red to blue-purple. The anthocyanin biosynthetic pathway is well understood. The enzymes required in this pathway mainly include chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3 -hydroxylase (F3 H), flavonoid 3 ,5 -hydroxylase (F3 5 H), dihydroflavonol-4-reductase (DFR) and anthocyanidin synthase (ANS) [2]. CHS is the key enzyme in the early stage of anthocyanin biosynthesis, and DFR and ANS are key enzymes in the late stage of anthocyanin synthesis, both of which play an important role in the accumulation of anthocyanins [3,4]. Anthocyanin biosynthesis is regulated by transcription factors that can recognize specific DNA motifs in the promoter of structural genes [5]

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