目的 在全基因组水平鉴定蒙古黄芪生物调控因子（Growth-regulating factors，GRF）转录因子家族成员，开展系统的生物信息学分析，并探究该家族基因在黄芪不同生长期的表达模式，为研究GRF转录因子调控黄芪次生代谢的分子机制提供基础。方法 通过生物信息学方法鉴定黄芪GRF转录因子家族，对其编码蛋白的理化特性、保守结构域组成、潜在亚细胞定位、系统发育关系及启动子区顺式调控元件进行预测分析，利用转录组数据研究GRF基因在黄芪不同生长期的表达模式，并与黄芪根中黄酮皂苷含量数据进行基因表达与代谢物积累的相关性评估。结果 蒙古黄芪基因组共鉴定获得9个GRF转录因子家族成员，分别定位在7条染色体，聚类为5个亚家族。AmGRF蛋白氨基酸残基数目在320～642 aa，均表现为亲水性特征，不存在跨膜结构域及信号肽序列，亚细胞定位均在细胞核。系统进化分析显示，AmGRF转录因子与大豆GRF亲缘关系较近。AmGRF基因启动子区存在大量与光信号响应、激素信号转导、生长发育及逆境胁迫应答相关的顺势作用元件。转录组数据表明，AmGRF基因各成员在不同生长期根部组织的表达模式存在较大差异，表明各家族成员可能在黄芪不同生长阶段中发挥特异的调控作用。基因表达与黄芪黄酮皂苷含量相关性分析显示，AmGRF3和AmGRF4与黄芪根中黄酮代谢调控相关，AmGRF5、AmGRF6和AmGRF8与黄芪根中皂苷代谢调控相关。结论 在全基因组水平对蒙古黄芪GRF基因家族进行鉴定和生信分析，通过黄芪不同生长期根中皂苷黄酮含量测定和GRF基因表达数据，鉴定出与黄芪黄酮皂苷代谢相关的GRF转录因子，为深入解析该家族调控蒙古黄芪生长发育及次生代谢的生物学功能提供关键靶点。

To identify the growth-regulating factors (GRF) transcription factor family at the whole genome level of Astragalus membranaceus var. mongholicus by bioinformatics methods, and analyze its expression pattern at different growth stages, providing a basis for studying the molecular mechanism of GRF transcription factors regulating the secondary metabolism of A. membranaceus var. mongholicus. Methods The GRF transcription factor family members of A. membranaceus var. mongholicus were identified, and their protein physicochemical properties, protein conserved motifs, subcellular localization, phylogenetic analysis, and cis-acting element prediction were analyzed by bioinformatics methods. The transcriptome data were used to study the expression pattern of the GRF gene in different growth stages of A. membranaceus var. mongholicus. Results A total of nine GRF transcription factors were identified, which were located on seven chromosomes and clustered into five subfamilies. The amino acid number of AmGRFs was 320—642 aa. They were all hydrophilic proteins without transmembrane structure and signal peptide structure, and all of which are located in the nucleus. Phylogenetic analysis showed that AmGRF transcription factors were closely related to soybean GRFs. There were a large number of cis-acting elements related to light signal response, hormone signal transduction, growth and development, and stress response to adverse conditions in the promoter region of AmGRF genes. Transcriptome data showed that the expression patterns of AmGRF genes in root tissues at different growth stages were quite different. This indicated that each family member may play a specific regulatory role in different growth stages of A. membranaceus var. mongholicus. Correlation analysis between gene expression and flavonoid and saponin content of A. membranaceus var. mongholicus showed that AmGRF3 and AmGRF4 were related to the regulation of flavonoid metabolism, while AmGRF5, AmGRF6 and AmGRF8 were related to the regulation of saponin metabolism in A. membranaceus var. mongholicus root. Conclusion The identification and bioinformatics analysis of the GRF gene family of A. membranaceus var. mongholicus were conducted at the whole-genome level. Based on the determination of saponin and flavonoid contents in the roots of A. membranaceus var. mongholicus at different growth stages and the expression data of GRF genes, the GRF transcription factors related to the metabolism of flavonoid saponins in A. membranaceus var. mongholicus were identified, providing key targets for in-depth analysis of the biological functions of this family in regulating the growth and development as well as secondary metabolism of A. membranaceus var. mongholicus.

R286.12

山西省科技成果转化引导专项（202304021301059）；山西省科技重大专项计划“揭榜挂帅”项目（202405071301030）；山西省现代农业中药材产业技术体系建设专项（2025）；山西农业大学2025年度“特”“优”农业高质量发展科技支撑工程项目“恒山黄芪微生物接种播种技术示范”（TYGC25，79-81）