青阳参C21甾体类生物合成关键基因挖掘  

Mining of Key Genes for Biosynthesis of C21 Steroids of Cynanchum otophyllum

作　　者：耿应敏 周兴乾 吴丽丽 张体操 郑兰平 GENG Yingmin;ZHOU Xingqian;WU Lili;ZHANG Ticao;ZHENG Lanping(College of Chinese Materia Medica,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南中医药大学中药学院,昆明650500

出　　处：《中国药学杂志》2025年第5期447-457,共11页Chinese Pharmaceutical Journal

基　　金：云南省基础研究项目资助(202301AT070254);云南省应用基础研究中医联合专项资助(202101AZ070001-056)。

摘　　要：目的 解析青阳参C_(21)甾体类化合物的生物合成途径。方法 本研究利用代谢组学和转录组学技术,比较分析青阳参根、茎、叶中C_(21)甾体类化合物的相对含量,再通过京都基因与基因组百科全书(KEGG)数据库注释,筛选与C_(21)甾体类生物合成相关的基因。最后通过实时荧光定量聚合酶链式反应(quantitative real-time PCR, qRT-PCR)对部分差异表达基因(differentially expressed genes, DEGs)进行基因表达验证。结果 青阳参苷元在根中显著上调,且在根中的相对含量约为叶中的73.10倍,约为茎中的19.05倍。转录组学分析发现,青阳参中有269条DEGs注释到了C_(21)甾体类生物合成相关通路。通过分析各比较组间注释到的DEGs,在C_(21)甾体类相关合成通路中筛选出了18个关键酶,且由87条基因编码,其中AACT等酶是其生物合成途径上游阶段的关键酶。进一步对其中8条DEGs进行qRT-PCR验证,其表达趋势与对应的转录组数据结果一致。结论 本研究系统解析了青阳参中C_(21)甾体类化合物的生物合成途径,筛选出多个关键酶及编码基因,丰富了青阳参的组学数据。这些发现为进一步研究青阳参C_(21)甾体类化合物的生物合成机制奠定了基础。OBJECTIVE To investigate the biosynthetic pathway of C_(21) steroidal compounds in Cynanchum otophyllum Schneid..METHODS Metabolomics and transcriptomics were used to compare and analyze the relative contents of C_(21) steroids in the roots,stems and leaves of C.otophyllum.Then,the genes related to C_(21) steroid biosynthesis in C.otophyllum were screened through Kyoto Encyclopedia of Genes and Genomes(KEGG)database annotation.Finally,some differentially expressed genes(DEGs)were verified by quantitative real-time PCR.RESULTS The qingyangshengenin content in the roots was significantly up-regulated,with the relative qingyangshengenin content in the roots being approximately 73.10 times higher than that in the leaves and 19.05 times higher than that in the stems.Transcriptomic analysis revealed that 269 DEGs annotated C_(21) steroidal biosynthetic pathways.By analyzing the DEGs annotated between the comparison groups,18 key enzymes were screened out in the C_(21) steroidal synthesis pathway,which were encoded by 87 genes,among which AACT and other enzymes were the key enzymes in the upstream stage of the biosynthesis pathway.Quantitative real-time PCR was performed to verify the expression trend of eight DEGs,which was consistent with the corresponding transcriptome data.CONCLUSION The biosynthetic pathway of C_(21) steroidal compounds is systematically analyzed in this study,and several key enzymes and coding genes are screened,which enrich the omics data of C.otophyllum.These findings lay a foundation for further study on the biosynthesis mechanism of C_(21) steroid compounds of C.otophyllum.

关 键 词：青阳参 青阳参苷元 代谢组学 转录组学 生物合成途径 

分 类 号：R29[医药卫生—民族医学] R931.6[医药卫生—中医学]