| 摘要: |
| 【 目的】明确复色菊花青素生物合成途径的关键基因，为其花斑形成的分子机制研究奠定基础。【 方 法】以复色菊‘花洒摇滚’为研究对象，根据英国皇家园艺学会比色卡（RHSCC）结合色差仪对其花色进行数据化分 析，并测定其总花青素和总黄酮含量；将同一花瓣不同颜色区域分别进行不同时期转录组测序及分析，以P且 log2|fold change|≥1.5作为条件筛选差异表达基因（DEGs），对DEGs进行GO及KEGG功能富集分析；根据注释结果 筛选并鉴别花青素生物合成相关结构基因与MYB转录因子；用荧光定量PCR（RT-qPCR）技术进行验证。【 结果】‘花洒摇滚’系复色菊花品种，花色为紫红色系与白色系，S2 时期是紫色花斑区域呈色的关键时期，花青素含量及其分 布是影响紫色花斑形成的主要因素。经转录组分析，在S2时期PS2和WS2共有4 779个差异表达基因，相对于白色区域，2 506个差异表达基因在紫色区域上调表达，2 273个差异表达基因在紫色区域下调表达。GO富集分析结果显 示，S2时期有3 533个DEGs注释到生物过程、分子功能、细胞组分3种功能类型，其中生物过程中的细胞过程注释到1 629个差异表达基因。KEGG富集分析结果表明，DEGs显著富集在代谢、遗传信息处理、生物系统等信号通路。根据KEGG注释结果确定类黄酮合成、花青素合成途径为花色生物合成的关键途径；结合2个代谢途径及关键时期筛选 得到与花色合成途径相关的关键基因为F3’H、FLS、DFR、ANS、MYB1、MYB3，RT-qPCR验证结果与转录组分析结 果一致。【 结论】F3’H、DFR、ANS、MYB1、MYB3是正向调控花青素积累形成紫色花斑的关键基因。 |
| 关键词: 复色菊 转录组学 花色 RT-qPCR |
| DOI：10. 13207/j. jnwafu. 2026. 05. 008 |
| 分类号: |
| 基金项目:河北省现代农业产业技术体系花卉创新团队建设专项（HBCT2024200201） |
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| Exploration of key genes in anthocyanin biosynthesis in the multicolored Chrysanthemum morifolium |
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XU Yingxin, ZHANG Ce, LIU Yuguang, ZHOU Xiaohui, SHI Baosheng
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College of Forestry，Hebei Agricultural University，Baoding，Hebei 071000，China
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| Abstract: |
| 【 Objective】This study aims to identify key genes involved in the anthocyanin biosynthesis pathway of multicolored Chrysanthemum morifolium，thereby providing a molecular basis for elucidating the mechanisms underlying floral pigmentation patterning.【Method】Takes the multicolored Chrysanthemum morifolium ‘Huasayaogun’ as research object，floral coloration was quantified by using the Royal Horticultural Society Colour Chart（RHSCC） in combination with a colorimeter，and the total anthocyanin and flavonoid contents were determined. Comparative transcriptome sequencing and analysis were performed on distinct color sectors of the same ray florets across different developmental stages. Genes with P<0.05 and log2 |fold change|≥1. 5 were identified as differentially expressed genes（DEGs） and subsequently subjected to GO（Gene Ontology） and KEGG（Kyoto Encyclopedia of Genes and Genomes） functional enrichment analyses. Structural genes and MYB transcription factors associated with anthocyanin biosynthesis were screened and identified based on functional annotation results，and quantitative real-time PCR（RT-qPCR） was used for validation.【Result】The multicolored chrysanthemum cultivar‘ Huasayaogun’ was characterized by a combination of purple-red and white color sectors. Stage S2 is the critical phase for pigmentation in the purple variegated regions，where anthocyanin accumulation and spatial distribution are the key determinants of purple spot formation. Transcriptomic analysis revealed a total of 4 779 differentially expressed genes（DEGs） between the purple（PS2） and white （WS2） sectors during Stage S2，with 2 506 DEGs upregulated and 2 273 DEGs downregulated in the purple sectors in comparison with the white sectors. The GO enrichment analysis revealed that during the S2 stage，3 533 differentially expressed genes（DEGs） were annotated to three functional categories：biological process，molecular function，and cellular component. Within the biological process category，the subcategory cellular process was annotated with 1 629 genes. KEGG pathway enrichment analysis revealed that DEGs were significantly enriched in major functional categories，including metabolism，genetic information processing，and organismal systems，based on the KEGG database classification hierarchy. Based on integrated analysis of KEGG annotations，flavonoid biosynthesis and anthocyanin biosynthesis pathways were identified as key pathways for floral pigmentation. By analyzing the flavonoid and anthocyanin biosynthesis pathways during the critical stage of floral
pigmentation，six key genes：F3’H，FLS，DFR，ANS，MYB1，and MYB3，were identified as being closely associated with pigment synthesis. RT-qPCR validation results were consistent with the transcriptomic analysis.
【Conclusion】F3’H，DFR，ANS，MYB1，and MYB3 were identified as key positive regulators driving anthocyanin accumulation and subsequent purple spot formation. |
| Key words: multicolored Chrysanthemum morifolium transcriptomics flower color RT-qPCR |
