紫苏PfMYB80转录因子正向调控花青素的生物合成

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

摘要/Abstract

摘要：

目的 R2R3-MYB转录因子主要参与调控类黄酮及花青素等次生代谢产物生物合成途径。验证其在紫苏（Perilla frutescens （L.） Britt.）花青素生物合成中的功能，为揭示R2R3-MYB转录因子在调控植物花青素合成中的作用奠定基础。方法基于生物信息学技术鉴定了紫苏基因组数据库的R2R3-MYB转录因子，并对其理化性质、系统进化、染色体定位、启动子顺式作用元件进行预测分析；筛选到可能参与调控紫苏花青素生物合成的R2R3-MYB成员，应用RT-qPCR分析该转录因子在紫苏叶片中的表达模式，克隆叶片中高表达的PfMYB80基因编码序列，探讨PfMYB80对紫苏花青素的合成调控作用及对红、蓝光胁迫的响应机制。结果共鉴定到186个R2R3-MYB成员，系统进化分析显示，紫苏PfMYB80、PfMYB146与拟南芥S6亚组调控植物花青素合成的基因亲缘关系最近，推测其可能参与调控花青素的合成。启动子顺式作用元件分析结果表明，紫苏R2R3-MYB基因启动子区含有光胁迫响应元件；RT-qPCR分析结果显示，PfMYB80基因在紫苏叶片不同发育时期的表达量逐渐升高，与花青素的合成趋势一致，推测其可能参与花青素的生物合成，该蛋白定位于细胞核；PfMYB80与花青素合成相关结构基因表达谱分析结果表明，紫苏LDOX基因的表达量与PfMYB80的表达趋势一致，且与紫苏花青素积累趋势一致，推测PfMYB80转录因子可能通过直接调控LDOX基因的转录表达来调节花青素的合成。通过转基因烟草的功能分析发现，PfMYB80响应蓝光诱导，并正向调控花青素的合成。结论紫苏PfMYB80转录因子正向调控花青素的生物合成，且响应光应答，蓝光下过表达PfMYB80烟草植株更有利于花青素的积累，同时可提高SOD 酶活性，降低POD酶活性和MDA含量。

关键词: 紫苏, R2R3-MYB转录因子, PfMYB80转录因子, 花青素, 光胁迫响应, 功能分析

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

李锐, 胡婷, 陈树溦, 王尧, 王计平. 紫苏PfMYB80转录因子正向调控花青素的生物合成[J]. 生物技术通报, 2025, 41(6): 243-255.

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.

图/表 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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