大黄藤中黄藤素生物合成途径关键酶基因的筛选  

Screening of key enzyme genes on the palmatine biosynthetic pathway in Fibraurea recisa

作　　者：周兴乾 耿应敏 张体操 郑兰平 ZHOU Xing-qian;GENG Ying-min;ZHANG Ti-cao;ZHENG Lan-ping(College of Chinese Materia Medica and Yunnan Key Laboratory of Southern Medicinal Utilization,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南中医药大学中药学院暨云南省南药可持续利用重点实验室,云南昆明650500

出　　处：《药学学报》2024年第6期1873-1882,共10页Acta Pharmaceutica Sinica

基　　金：云南省应用基础研究中医联合专项(202101AZ070001-056);云南省基础研究项目(202301AT070254);云南省南药可持续利用重点实验室开放课题(202005AG070157).

摘　　要：黄藤素是大黄藤的主要功效成分,是典型的小檗碱类异喹啉类生物碱,具有广泛的抗炎、抗菌活性。本研究运用代谢组学与转录组学研究,筛选了与黄藤素合成显著相关的差异表达基因(DEGs),并对其中8个DEGs进行了实时荧光定量PCR(qRT-PCR)验证。代谢组学研究共检测到106种生物碱,其中包含23种异喹啉类生物碱。黄藤素在根与叶的对比组中位居差异代谢物差异倍数前十位,其在根中的相对含量约是叶中的47.5倍。通过转录组学研究,共188条基因注释到异喹啉类生物碱的生物合成途径上,其中36条基因具有显著差异,根与叶的对比组包含33条DEGs,30条DEGs在黄藤素的生物合成途径上。最后,通过代谢组学与转录组学关联分析,筛选到4条基因序列的表达模式与黄藤素具有显著相关性。qRT-PCR实验结果表明8个DEGs的表达趋势均与转录组结果一致。本研究不仅丰富了大黄藤的组学数据,也为黄藤素的生物合成研究奠定了基础,并进一步为其他异喹啉类生物碱生物合成途径上关键酶基因的解析提供了参考。Palmatine,the main effective ingredient of Fibraurea recisa,is a typical berberine isoquinoline alkaloid with extensive anti-inflammatory and antibacterial activities.In this work,the studies of metabolomics and transcriptomics were utilized to detect differentially expressed genes(DEGs)that are significantly associated with the synthesis of palmatine.In addition,eight of these DEGs were verified by quantitative real-time PCR(qRTPCR).A total of 106 alkaloids were detected in the metabolomics study,including 23 isoquinoline alkaloids.Palmatine ranked in the top ten of differential metabolites in the group of root vs leaf,and its relative content in root was about 47.5 times higher than that in leaf.In the transcriptomics study,a total of 188 genes were annotated to the pathway of isoquinoline alkaloid biosynthesis.Among them,there were 36 DEGs were significantly different.In the comparison group of root and leaf,a total of 33 DEGs were significantly different,and 30 DEGs were annotated on the biosynthetic pathway of palmatine.Finally,the results of the correlation analysis between metabolomics and transcriptomics showed that the expression patterns of four gene sequences were screened to be significantly correlated with palmatine.The results of qRT-PCR experiments showed that the expression trends of eight DEGs were consistent with the results of transcriptomic.This study not only enriched the omics data of F.recisa,but also established the foundation for the study of the synthetic biology of palmatine.It further provided a reference for the analysis of the key enzyme genes on the biosynthetic pathway of other isoquinoline alkaloids.

关 键 词：大黄藤 多组学 生物合成途径 黄藤素 异喹啉类生物碱 

分 类 号：R931[医药卫生—生药学]