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【目的】研究青蒿素生物合成基因在不同发育时期和不同组织中的表达规律,探讨青蒿素生物合成的时空调节机制。【方法】利用实时荧光定量PCR技术定量追踪分析青蒿素主要及协同合成途径基因在盛叶期、开花前、开花后不同的发育时期,以及在根、茎、叶、花不同组织中的表达模式。【结果】各基因的表达水平在6月份时最低,随后开始上升,在8月份(开花前)达到高峰,与6月份比较,转录水平显著提高3～15倍(均P<0.01),其中青蒿素特异合成酶基因ADS和CYP71AV1上升幅度最大,为最低水平的12和15倍,9月份(开花后)则呈下降趋势。处于盛花期的青蒿在根、茎、叶、花各个组织中都能检测到青蒿素合成基因的表达,叶片中ADS基因mRNA水平较其他组织显著升高2倍左右(均P<0.01),显示组织表达特异性。【结论】青蒿素生物合成基因的发育表达模式与青蒿素的合成规律一致。ADS和CYP71AV1基因的表达在青蒿素生物合成过程中可能起关键作用。
【Objective】 The aim of this study was to investigate the expression pattern of artemisinin biosynthesis genes at different developmental stages and in different tissues and to explore the spatial and temporal regulation mechanism of artemisinin biosynthesis. 【Method】 Real-time quantitative PCR was used to quantitatively analyze the expression of major and synergistic synthetic pathway genes of artemisinin at different developmental stages, such as the stage of blooming, pre-flowering, post-flowering, and in different tissues of roots, stems, leaves and flowers mode. 【Result】 The results showed that the expression level of each gene was the lowest in June and then began to rise. The peak reached the peak in August before flowering, and the transcription level increased 3 ~ 15 times (all P <0.01) The genes of artemisinin-specific synthase (ADS) and CYP71AV1 increased the most at the 12th and 15th times of the lowest levels, and declined in September (after flowering). Artemisia annua L. in full flowering stage could detect the expression of artemisinin synthetic gene in all tissues of roots, stems, leaves and flowers, and the mRNA level of ADS gene in leaves was significantly increased about 2 times (P <0.01) , Showing tissue expression specificity. 【Conclusion】 The pattern of development and expression of artemisin biosynthesis genes is consistent with that of artemisinin. The expression of ADS and CYP71AV1 genes may play a key role in artemisin biosynthesis.