Screening of key enzyme genes on the palmatine biosynthetic pathway in Fibraurea recisa
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(qRT-PCR).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.
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