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【目的】探讨青蒿细胞质与质体中的萜类合成途径的相互关系。【方法】用甲羟戊酸(MVA)途径抑制剂洛伐他汀、脱氧木酮糖磷酸(DXP)途径抑制剂磷甘霉素及类固醇合成抑制剂咪康唑对青蒿苗进行处理,对途径终产物——β-胡萝卜素和生育酚的生成以及细胞质内的MVA途径关键酶基因(HMGR、FPS、ADS、CYP71AV1、CPR、SQS)和质体内的DXP途径关键酶基因(DXS、DXR)的转录表达水平的动态变化进行了定量依时分析。【结果】洛伐他汀对MVA途径基因(HMGR、FPS、ADS、CYP71AV1、CPR、SQS)和DXP途径基因(DXS、DXR)的转录均有抑制作用,但对DXP途经终产物(β-胡萝卜素和生育酚)的合成无显著影响,叶片也并未出现白化。磷甘霉素处理后对DXS、DXR转录和β-胡萝卜素、生育酚合成有持续抑制作用,但MVA途径基因转录在3h瞬时提高,青蒿苗同时出现短根和叶片白化现象。添加咪康唑后青蒿素合成基因(ADS、CYP71AV1、CPR)的转录水平分别提高11、5和3倍。【结论】表明青蒿2条萜类合成途径之间存在物质交流,且流动方向可能以叶绿体向胞浆流动为主,抑制青蒿素合成竞争途径可调节细胞代谢向青蒿素合成方向移动。
【Objective】 The aim of this study was to investigate the relationship between terpenoids synthesis pathway in Artemisia annua cytoplasm and plastid. 【Method】 Artemisia annua L. plants were treated with lovastatin, the inhibitor of mevalonate (MVA) pathway, the inhibitor of DXX pathway and miconazole, a steroid synthesis inhibitor, The production of β-carotene and tocopherol as well as the key enzyme genes (DXS, DXR) of the key enzymes of the MVA pathway (HMGR, FPS, ADS, CYP71AV1, CPR and SQS) The dynamic changes of transcriptional expression level were quantitatively analyzed on time. 【Results】 Lovastatin inhibited the transcription of MVA pathway genes (HMGR, FPS, ADS, CYP71AV1, CPR, SQS) and DXP pathway genes (DXS, DXR) And tocopherol) had no significant effect on the synthesis of leaves did not appear albinism. However, the transcription of MVA pathway was transiently increased at 3h, and the short roots and leaf albino were also observed in A. annua L. plants. The transcription of artemisinin-synthesizing genes (ADS, CYP71AV1, CPR) increased by 11, 5 and 3 fold, respectively, after addition of miconazole. 【Conclusion】 The results showed that there was material exchange between two terpenoids in Artemisia annua L., and the flow direction was probably chloroplast to cytoplasm. Inhibition of artemisinin biosynthesis pathway could regulate cell metabolism toward artemisinin synthesis.