Positive Regulation of Anthocyanin Biosynthesis by PfMYB80 Transcription Factor in Perilla frutescens

doi:10.13560/j.cnki.biotech.bull.1985.2024-1211

Abstract

Abstract:

Objective The R2R3-MYB transcription factor is mainly involved in regulating the biosynthesis pathways of secondary metabolites such as flavonoids and anthocyanins. To verify its function in the anthocyanin biosynthesis of perilla (Perilla frutescens (L.) Britt.) would lay the foundation for elucidating the role of R2R3-MYB transcription factors in regulating plant anthocyanin synthesis. Method This study employed bioinformatics analysis to identify the R2R3-MYB transcription factors across the entire genome of perilla, and to predict their physicochemical properties, phylogenetic evolution, chromosome localization, and cis-acting elements of the promoter. R2R3-MYB members potentially involved in regulating the biosynthesis ofperilla anthocyanin were screened through correlation analysis, and the highly expressed PfMYB80 gene coding sequence in leaves was cloned to explore the regulatory effect of PfMYB80 on anthocyanin synthesis in perilla and its response mechanism to red and blue light stress. Result A total of 186 R2R3-MYB members were identified. Phylogenetic analysis revealed that PfMYB80 and PfMYB146 from perilla are most closely related to the genes in subgroup S6 of Arabidopsis thaliana, which are known to regulate anthocyanin synthesis in plants, suggesting their potential involvement in the regulation of anthocyanin synthesis. Analysis of promoter cis-acting elements indicated that the promoter regions of Perilla R2R3-MYB genes contain light stress-responsive elements. RT-qPCR results showed that the expression of the PfMYB80 gene gradually increases during different developmental stages of Perilla leaves, consistent with the trend of anthocyanin synthesis, suggesting its possible role in anthocyanin biosynthesis. This protein is localized in the nucleus. Expression profile analysis of PfMYB80 and anthocyanin synthesis-related structural genes revealed that the expression of the LDOX gene in Perilla aligns with that of PfMYB80 and matches the trend of anthocyanin accumulation, indicating that the PfMYB80 transcription factor might regulate anthocyanin synthesis by directly controlling the transcriptional expression of the LDOX gene. Functional analysis through transgenic tobacco demonstrated that PfMYB80 responds to blue light induction and positively regulates anthocyanin synthesis. Conclusion The PfMYB80 transcription factor in Perillapositively regulate the biosynthesis of anthocyanins and respond to light response. Overexpression of PfMYB80 in tobacco under blue light treatment increases anthocyanin accumulation and enhances SOD enzyme activity and reduces POD enzyme activity and MDA content.

Key words: Perilla?frutescens, R2R3-MYB transcription factor, PfMYB80 transcription factor, anthocyanin, light stress response, functional analysis

LI Rui, HU Ting, CHEN Shu-wei, WANG Yao, WANG Ji-ping. Positive Regulation of Anthocyanin Biosynthesis by PfMYB80 Transcription Factor in Perilla frutescens[J]. Biotechnology Bulletin, 2025, 41(6): 243-255.

Figures/Tables 2

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引物名称 Primer name	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）	长度 Length （bp）
PfMYB80-qPCR	GAACGCTTCTTCATCTTCATCTT	TCCTCTAATAAACCACGCAATCC	226
PfMYB146-qPCR	ATATCCAAGAAACCGCCGTCTGC	TGTGAGTTAGAGAAGGTCCGAGGTC	101
Pf4CL-qPCR	TGGAGGATGCTGTCAGGGCTAAG	CTCAAACGGCTCTTTCGCAAACG	116
PfF3H-qPCR	GCCTACGACCAATTCAGCAGTGAG	TCACACGCCGCCACTATCCTC	107
PfF3′H-qPCR	GACAATAATGGTGAGGGAGGGAAGC	CCGTGCTAGATGTCGTGTCTGTTC	97
PfDFR-qPCR	TGACGGCATTCACAAGGACATAGC	GCAGCTCTCAATGGCACCTCTATAC	118
PfUFGT-qPCR	TGAAAGACTACGCCGTGAAATCCC	GTTCCAGCCGCAGTGAGTGAC	143
PfLDOX-qPCR	GTTGGTGGTGCGGAGGATCTAATC	CATGTTGTGGAGGATGAAGGTGAGG	132
PfActin-qPCR	AGACCTTCAATGTGCCAGCCA	CACGACCAGCAAGATCCAACC

引物名称 Primer name	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）	长度 Length （bp）
PfMYB80-qPCR	GAACGCTTCTTCATCTTCATCTT	TCCTCTAATAAACCACGCAATCC	226
PfMYB146-qPCR	ATATCCAAGAAACCGCCGTCTGC	TGTGAGTTAGAGAAGGTCCGAGGTC	101
Pf4CL-qPCR	TGGAGGATGCTGTCAGGGCTAAG	CTCAAACGGCTCTTTCGCAAACG	116
PfF3H-qPCR	GCCTACGACCAATTCAGCAGTGAG	TCACACGCCGCCACTATCCTC	107
PfF3′H-qPCR	GACAATAATGGTGAGGGAGGGAAGC	CCGTGCTAGATGTCGTGTCTGTTC	97
PfDFR-qPCR	TGACGGCATTCACAAGGACATAGC	GCAGCTCTCAATGGCACCTCTATAC	118
PfUFGT-qPCR	TGAAAGACTACGCCGTGAAATCCC	GTTCCAGCCGCAGTGAGTGAC	143
PfLDOX-qPCR	GTTGGTGGTGCGGAGGATCTAATC	CATGTTGTGGAGGATGAAGGTGAGG	132
PfActin-qPCR	AGACCTTCAATGTGCCAGCCA	CACGACCAGCAAGATCCAACC

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