目的 克隆鸡血藤Spatholobus suberectus查耳酮合成酶基因（chalcone synthase，CHS）SsCHS1，开展生物信息学及基因功能研究，以期解析其在黄酮生物合成中的作用。方法 以鸡血藤cDNA为模板，借助RT-PCR技术克隆SsCHS1，对该基因的序列特征、蛋白结构与功能等开展系统的生物信息学解析。构建SsCHS1过表达载体，遗传转化本氏烟草，测定转基因植株中总黄酮含量及黄酮合成途径中关键酶基因的表达水平。结果 SsCHS1基因全长1 176 bp，其编码蛋白由391个氨基酸组成，理论相对分子质量为42 825.35，等电点为6.10，属于亲水性蛋白；与豆科植物甘草、大豆等物种的CHS蛋白具有较高同源性。启动子分析表明，SsCHS1富集了激素响应、光响应、胁迫响应及转录因子结合位点相关的多种顺式作用元件。功能分析显示，过表达SsCHS1可显著促进转基因烟草中黄酮的积累，并上调黄酮生物合成途径中关键酶基因的表达水平。结论 鸡血藤SsCHS1基因能够促进黄酮的生物合成，该基因的发掘为深入解析鸡血藤活性成分的合成机制提供了重要的理论依据与候选基因。

Objective To clone the chalcone synthase gene SsCHS1 from Spatholobus suberectus and conduct bioinformatics and functional analyses, in order to elucidate its role in flavonoid biosynthesis. Methods Using cDNA from S. suberectus as a template, SsCHS1 was cloned by RT-PCR. Systematic bioinformatics analyses were performed on the sequence characteristics, protein structure, and function of the gene. An SsCHS1 overexpression vector was constructed and genetically transformed into Nicotiana benthamiana. The total flavonoid content and the expression levels of key enzyme genes in the flavonoid biosynthesis pathway in transgenic plants were determined. Results The full-length SsCHS1 gene was 1 176 bp, encoding a protein consisting of 391 amino acids, with a theoretical relative molecular mass of 42 825.35, an isoelectric point of 6.10, and was characterized as a hydrophilic protein. It showed high homology with CHS proteins from leguminous plants such as Glycyrrhiza uralensis and Glycine max. Promoter analysis revealed that SsCHS1 contains multiple cis-acting elements associated with hormone response, light response, stress response, and transcription factor binding sites. Functional analysis demonstrated that overexpression of SsCHS1 significantly promoted flavonoid accumulation in transgenic tobacco and upregulated the expression levels of key enzyme genes in the flavonoid biosynthesis pathway. Conclusion The SsCHS1 gene from S. suberectus promotes flavonoid biosynthesis. The identification of this gene provides an important theoretical basis and a candidate gene for further elucidation of the biosynthetic mechanisms of active components in S. suberectus.

R282.12

国家重点研发计划项目（2024YFC3506703）；广西中医药适宜技术开发与推广项目（GZSY2025005）；广西自然科学基金项目（2025GXNSFAA069354）