目的 研究秃叶黄檗Phellodendron chinense var. glabriusculum不同组织部位异喹啉类生物碱成分含量差异及生物合成相关基因。方法 采用高效液相色谱法对秃叶黄檗根皮、茎皮、叶进行含量测定，结合转录组测序筛选关键基因，探讨秃叶黄檗异喹啉生物碱合成机制。结果 秃叶黄檗中异喹啉类生物碱在根皮、茎皮与叶中含量差异显著。转录组测序组装获得85 908条单基因（Unigenes），其中49 114（57.17%）条被成功注释，参与异喹啉类生物碱成分合成的38个差异表达基因被成功鉴定，并与小檗碱等异喹啉类成分进行相关性分析，显示8个基因与小檗碱含量高度正相关，其中主要包括CNMT、NCS、TDC、BBE、TYNA等基因，这些基因可能对于小檗碱基因的合成积累具有重要影响作用，对差异表达基因进行qRT-PCR，结果与转录组数据表达模式具有一致性。结论 揭示了秃叶黄檗不同组织部位异喹啉生物碱合成通路相关基因的显著差异表达模式，为进一步研究异喹啉生物碱生物合成调控分子机制提供线索。

Objective To study the content difference of isoquinoline alkaloids in different tissue parts of Phellodendron chinense var. glabriusculum and the genes related to biosynthesis Methods High performance liquid chromatography (HPLC) was used to determine the contents of root bark, stem bark and leaves of Amur barberberry, and transcriptome sequencing was used to screen key enzyme genes to explore the synthesis mechanism of isoquinoline alkaloids in P. chinense var. glabriusculum. Result The contents of isoquinoline alkaloids in root bark, stem bark and leaves were significantly different. 85 908 Unigenes were obtained by transcriptome sequencing, of which 49 114 (57.17%) were successfully annotated. 38 differentially expressed genes that catalyzed the production of isoquinoline alkaloids were successfully identified, and correlation analysis was conducted with isoquinoline components such as berberine. Eight genes were highly positively correlated with berberine content, including CNMT, NCS, TDC, BBE, TYNA and other genes, which may have important effects on the synthesis and accumulation of berberine genes. The expression pattern of differentially expressed genes was consistent with that of transcriptome data by qRT-PCR. Conclusion The significantly different expression patterns of isoquinoline alkaloid synthesis pathway related genes in different tissue parts of P. chinense var. glabriusculum were revealed, providing clues for further research on the molecular mechanism of isoquinoline alkaloid biosynthesis regulation.

R286.2

四川省中医药管理局项目（2023zd017）