植物学报

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ApWRKY33调控ApDFR基因表达影响红枫叶色变化的机制解析

张军1, 金小军1, 方彬3, 金松恒2, 董彬3*   

  1. 1 同创工程设计有限公司, 绍兴 312000; 2浙江农林大学暨阳学院, 诸暨 311800; 3浙江农林大学风景园林与建筑学院, 杭州 311300
  • 收稿日期:2025-09-16 修回日期:2025-11-06 出版日期:2025-12-08 发布日期:2025-12-08
  • 通讯作者: 董彬
  • 基金资助:
    浙江农林大学科研发展基金(No.2017FR016)

Mechanistic Analysis of ApWRKY33-mediated Regulation of ApDFR Expression in Leaf Color Variation of Acer palmatum

Jun Zhang1, Xiaojun Jin1, Bin Fang3, Songheng Jin2, Bin Dong3*   

  1. ¹Tongchuang Engineering Design Co., Ltd., Shaoxing 312000, China; ²Jiyang College of Zhejiang A&F University, Zhuji 311800, China; ³College of Landscape Architecture, Zhejiang A&F University, Hangzhou 311300, China
  • Received:2025-09-16 Revised:2025-11-06 Online:2025-12-08 Published:2025-12-08
  • Contact: Bin Dong

摘要: 红枫(Acer palmatum)是重要的彩叶植物, 常用于城镇绿化。光照是影响花青素合成的重要环境因子之一, 明确光照对红枫叶色的影响对其育种具有重要意义。通过自然条件调查和人工遮光处理, 发现遮光后红枫的叶片变绿, 花青素含量显著下降。转录组测序和荧光定量表达结果表明, 调控花青素合成通路基因(ApC4H、ApCHS、ApF3'H、ApDFRApANS)的表达显著下调; 同时, 鉴定出8个WRKY转录因子成员在遮光后也出现差异表达。通过酵母单杂交和双荧光素酶实验进一步发现, ApWRKY33转录因子能够直接结合在ApDFR基因的启动子上, 抑制ApDFR基因的表达。实验证实ApWRKY33能够响应光照, 遮光后通过抑制ApDFR的表达进而限制了红枫叶片花青素的合成, 使叶色变绿。研究结果将为解析红枫叶色变化的分子机理以及分子育种提供理论依据。

关键词: 彩叶植物,  红枫,  花青素,  基因调控

Abstract: INTRODUCTION: The formation of plant leaf color is associated with the content and proportion of various pigments, among which the accumulation of anthocyanin serves as a key factor determining red, purple, and similar hues in plant tissues or organs. Acer palmatum is an important color-leafed plant widely used in urban landscaping. Clarifying the impact of light on the leaf color of red maple holds significant importance for its breeding.  RATIONALE: Anthocyanin metabolism is dually regulated by genetic and environmental factors. Among environmental factors, light is considered a key element influencing anthocyanin synthesis or degradation. Different plant species exhibit varying responses to light, which constitutes an important molecular basis for the diversity of leaf coloration in colored-leaf plants. Anthocyanin biosynthesis involves a series of key enzymes, and these structural genes are regulated by transcription factors, including WRKY. However, the regulatory functions of WRKY transcription factors vary significantly across different species.  RESULTS: Through field observations and shading treatments, it was observed that the leaves of Acer palmatum turned green after shading, with a significant decrease in anthocyanin content. Transcriptome sequencing and quantitative real-time PCR (qRT-PCR) results indicated that the expression of key genes involved in the anthocyanin biosynthesis pathway (ApC4H, ApCHS, ApF3'H, ApDFR, and ApANS) was significantly downregulated. Concurrently, eight WRKY transcription factor members were identified as differentially expressed after shading. Further yeast one-hybrid and dual-luciferase assays demonstrated that the ApWRKY33 transcription factor can directly bind to the promoter of the ApDFR gene and inhibit its expression. The experiments confirmed that ApWRKY33 responds to light signals and represses anthocyanin biosynthesis by downregulating ApDFR under shading.  CONCLUSION: This study reveals the role of the ApWRKY33-ApDFR module in regulating leaf color in red maple. Future research should focus on elucidating how upstream light signals activate ApWRKY33 and employing techniques such as ChIP-seq to genome-widely identify its downstream targets, thereby refining the regulatory network. Protein interaction studies could help clarify the molecular basis of its functional specificity. Ultimately, the function of this module should be validated in vivo through genetic transformation systems, while molecular breeding techniques such as gene editing should be utilized to develop new cultivars of colored-leaf plants with stable leaf color traits and enhanced ornamental value.

Key words: colorful-leaf plants,  Acer palmatum,  anthocyanin,  gene regulation