目的 挖掘参与鸡血藤Spatholobus suberectus有效成分儿茶素和表儿茶素合成的关键候选基因及调控网络，为解析鸡血藤原花青素的生物合成路径及调控机制提供候选基因靶点。方法 以鸡血藤的嫩茎、嫩叶、老叶、木质部和韧皮部为材料，对不同部位进行转录组测序，并通过超高液相色谱串联质谱法（UPLC-MS/MS）技术分析儿茶素、表儿茶素黄酮类化合物在不同部位中含量，利用加权基因共表达网络分析（weighted gene co-expression network analysis，WGCNA）鉴定出与原花青素合成密切相关的关键模块和关键基因。另外，挑选8个基因进行qRT-PCR，验证转录组数据的可靠性。结果 鸡血藤各组织中儿茶素与表儿茶素的分布呈现显著差异，其中叶片中两者含量均显著低于茎部。通过WGCNA，将18 828个基因划分为15个共表达模块，发现turquoise、tan和brown模块与儿茶素、表儿茶素含量呈显著正相关（P＜0.05，r≥0.8）。京都基因与基因组百科全书（Kyoto encyclopedia of genes and 32genomes，KEGG）通路富集分析显示，这3个模块的基因主要富集于代谢相关通路。进一步分析表明，turquoise、tan和brown模块分别包含7个、4个和1个原花青素生物合成的结构基因，并鉴定出2个关键候选转录因子SsMYB1和SsMYB3。qRT-PCR验证结果显示，目标基因的表达变化趋势与转录组数据基本一致，证实了转录组分析结果的可靠性。结论 挖掘出的3个共表达模块和11个核心基因为深入解析鸡血藤原花青素合成的分子调控网络及关键基因的协同作用机制提供了重要研究基础。

Objective As potential active ingredients of Spatholobus suberectus, catechin and epicatechin play crucial roles in its medicinal properties.This study aimed to identify key candidate genes and regulatory networks involved in their biosynthesis, providing genetic targets for elucidating the biosynthetic pathway and regulatory mechanisms of proanthocyanidins in S. suberectus. Methods Young stems,young leaves, mature leaves, xylem, and phloem of S. suberectus were subjected to transcriptome sequencing. The contents of catechin, epicatechin, and other flavonoids in different tissues were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Weighted gene co-expression network analysis (WGCNA) was employed to identify key modules and genes closely associated with proanthocyanidin biosynthesis. Additionally, eight candidate genes were selected for qRT-PCR validation to confirm the reliability of the transcriptome data. Results The distribution of catechin and epicatechin varied significantly among different tissues,with both compounds showing significantly lower levels in leaves compared to stems. WGCNA analysis clustered 18 828 genes into 15 co-expression modules, among which the turquoise, tan, and brown modules exhibited highly significant positive correlations with catechin and epicatechin content (P < 0.05, r ≥ 0.8). KEGG enrichment analysis revealed that genes in these modules were primarily involved in metabolic pathways. Further analysis identified seven, four, and one structural genes related to proanthocyanidin biosynthesis in the turquoise, tan, and brown modules, respectively, along with two key candidate transcription factors, SsMYB1 and SsMYB3. The qRT-PCR validation confirmed that the expression trends of target genes were consistent with the transcriptome data, supporting the reliability of the findings. Conclusion The three co-expression modules and eleven core genes identified in this study provide a critical foundation for further exploration of the molecular regulatory network and synergistic mechanisms underlying proanthocyanidin biosynthesis in S. suberectus.

R286.12

广西自然科学基金项目（2023GXNSFAA026363）；2022年广西中医药大学引进博士科研启动基金项目（2022BS010）；中央本级重大增减支项目（2060302）；2025国家中医药管理局全国老药工传承工作室建设项目（国中医药人教函[2025]181号）；2025广西壮瑶药重点实验室课题（GXZYYZY2025-02）