目的 在全基因組水平鑒定蒙古黃芪生物調(diào)控因子（Growth-regulating factors，GRF）轉(zhuǎn)錄因子家族成員，開展系統(tǒng)的生物信息學(xué)分析，并探究該家族基因在黃芪不同生長期的表達(dá)模式，為研究GRF轉(zhuǎn)錄因子調(diào)控黃芪次生代謝的分子機(jī)制提供基礎(chǔ)。方法 通過生物信息學(xué)方法鑒定黃芪GRF轉(zhuǎn)錄因子家族，對其編碼蛋白的理化特性、保守結(jié)構(gòu)域組成、潛在亞細(xì)胞定位、系統(tǒng)發(fā)育關(guān)系及啟動子區(qū)順式調(diào)控元件進(jìn)行預(yù)測分析，利用轉(zhuǎn)錄組數(shù)據(jù)研究GRF基因在黃芪不同生長期的表達(dá)模式，并與黃芪根中黃酮皂苷含量數(shù)據(jù)進(jìn)行基因表達(dá)與代謝物積累的相關(guān)性評估。結(jié)果 蒙古黃芪基因組共鑒定獲得9個GRF轉(zhuǎn)錄因子家族成員，分別定位在7條染色體，聚類為5個亞家族。AmGRF蛋白氨基酸殘基數(shù)目在320～642 aa，均表現(xiàn)為親水性特征，不存在跨膜結(jié)構(gòu)域及信號肽序列，亞細(xì)胞定位均在細(xì)胞核。系統(tǒng)進(jìn)化分析顯示，AmGRF轉(zhuǎn)錄因子與大豆GRF親緣關(guān)系較近。AmGRF基因啟動子區(qū)存在大量與光信號響應(yīng)、激素信號轉(zhuǎn)導(dǎo)、生長發(fā)育及逆境脅迫應(yīng)答相關(guān)的順勢作用元件。轉(zhuǎn)錄組數(shù)據(jù)表明，AmGRF基因各成員在不同生長期根部組織的表達(dá)模式存在較大差異，表明各家族成員可能在黃芪不同生長階段中發(fā)揮特異的調(diào)控作用?；虮磉_(dá)與黃芪黃酮皂苷含量相關(guān)性分析顯示，AmGRF3和AmGRF4與黃芪根中黃酮代謝調(diào)控相關(guān)，AmGRF5、AmGRF6和AmGRF8與黃芪根中皂苷代謝調(diào)控相關(guān)。結(jié)論 在全基因組水平對蒙古黃芪GRF基因家族進(jìn)行鑒定和生信分析，通過黃芪不同生長期根中皂苷黃酮含量測定和GRF基因表達(dá)數(shù)據(jù)，鑒定出與黃芪黃酮皂苷代謝相關(guān)的GRF轉(zhuǎn)錄因子，為深入解析該家族調(diào)控蒙古黃芪生長發(fā)育及次生代謝的生物學(xué)功能提供關(guān)鍵靶點(diǎn)。

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

山西省科技成果轉(zhuǎn)化引導(dǎo)專項(xiàng)（202304021301059）；山西省科技重大專項(xiàng)計(jì)劃“揭榜掛帥”項(xiàng)目（202405071301030）；山西省現(xiàn)代農(nóng)業(yè)中藥材產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)（2025）；山西農(nóng)業(yè)大學(xué)2025年度“特”“優(yōu)”農(nóng)業(yè)高質(zhì)量發(fā)展科技支撐工程項(xiàng)目“恒山黃芪微生物接種播種技術(shù)示范”（TYGC25，79-81）