目的 研究禿葉黃檗Phellodendron chinense var. glabriusculum不同組織部位異喹啉類生物堿成分含量差異及生物合成相關(guān)基因。方法 采用高效液相色譜法對禿葉黃檗根皮、莖皮、葉進(jìn)行含量測定，結(jié)合轉(zhuǎn)錄組測序篩選關(guān)鍵基因，探討禿葉黃檗異喹啉生物堿合成機(jī)制。結(jié)果 禿葉黃檗中異喹啉類生物堿在根皮、莖皮與葉中含量差異顯著。轉(zhuǎn)錄組測序組裝獲得85 908條單基因（Unigenes），其中49 114（57.17%）條被成功注釋，參與異喹啉類生物堿成分合成的38個差異表達(dá)基因被成功鑒定，并與小檗堿等異喹啉類成分進(jìn)行相關(guān)性分析，顯示8個基因與小檗堿含量高度正相關(guān)，其中主要包括CNMT、NCS、TDC、BBE、TYNA等基因，這些基因可能對于小檗堿基因的合成積累具有重要影響作用，對差異表達(dá)基因進(jìn)行qRT-PCR，結(jié)果與轉(zhuǎn)錄組數(shù)據(jù)表達(dá)模式具有一致性。結(jié)論 揭示了禿葉黃檗不同組織部位異喹啉生物堿合成通路相關(guān)基因的顯著差異表達(dá)模式，為進(jìn)一步研究異喹啉生物堿生物合成調(diào)控分子機(jī)制提供線索。

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

四川省中醫(yī)藥管理局項目（2023zd017）