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茉莉酸甲酯和水杨酸对转基因丹参毛状根中丹参酮产量及生物合成基因表达的影响

Effects of methyl jasmonate and salicylic acid on tanshinone production and biosynthetic gene expression in transgenic Salvia miltiorrhiza hairy roots.

作者信息

Hao Xiaolong, Shi Min, Cui Lijie, Xu Chao, Zhang Yanjie, Kai Guoyin

机构信息

Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai, People's Republic of China.

出版信息

Biotechnol Appl Biochem. 2015 Jan-Feb;62(1):24-31. doi: 10.1002/bab.1236. Epub 2014 Jun 13.

Abstract

Tanshinone is a group of active diterpenes, which are widely used in the treatment of cardiovascular disease. In this study, methyl jasmonate (MJ) and salicylic acid (SA) were used to investigate their effects on tanshinone accumulation and biosynthetic gene expression in the hairy roots of geranylgeranyl diphosphate synthase (SmGGPPS) overexpression line (G50) in Salvia miltiorrhiza. High-performance liquid chromatography analysis showed that total tanshinone content in G50 was obviously increased by 3.10-fold (11.33 mg/g) with MJ at 36 H and 1.63 times (5.95 mg/g) after SA treatment for 36 H in comparison with their mimic treatment control. Furthermore, quantitative reverse-transcription PCR analysis showed that the expression of isopentenyl-diphosphate delta-isomerase (SmIPPI), SmGGPPS, copalyl diphosphate synthase (SmCPS), and kaurene synthase-like (SmKSL) increased significantly with MJ treatment. However, the expression of SmIPPI reached the highest level at 144 H, whereas those of SmGGPPS, SmCPS, and SmKSL only increased slightly with SA treatment. The two elicitor treatments suggested that tanshinone accumulation positively correlated to the expression of key genes such as SmGGPPS, SmCPS, and SmKSL. Meanwhile, the study also indicated that it was a feasible strategy to combine elicitor treatment with transgenic technology for the enhancement of tanshinone, which paved the way for further metabolic engineering of tanshinone biosynthesis.

摘要

丹参酮是一类活性二萜类化合物,广泛用于治疗心血管疾病。在本研究中,使用茉莉酸甲酯(MJ)和水杨酸(SA)来研究它们对丹参香叶基香叶基二磷酸合酶(SmGGPPS)过表达系(G50)毛状根中丹参酮积累和生物合成基因表达的影响。高效液相色谱分析表明,与模拟处理对照相比,G50中的总丹参酮含量在MJ处理36小时后明显增加3.10倍(11.33mg/g),SA处理36小时后增加1.63倍(5.95mg/g)。此外,定量逆转录PCR分析表明,MJ处理后异戊烯基二磷酸δ-异构酶(SmIPPI)、SmGGPPS、柯巴基二磷酸合酶(SmCPS)和贝壳杉烯合酶样(SmKSL)的表达显著增加。然而,SmIPPI的表达在144小时达到最高水平,而SmGGPPS、SmCPS和SmKSL的表达在SA处理后仅略有增加。这两种诱导处理表明,丹参酮积累与SmGGPPS、SmCPS和SmKSL等关键基因的表达呈正相关。同时,该研究还表明,将诱导处理与转基因技术相结合是提高丹参酮的可行策略,为丹参酮生物合成的进一步代谢工程铺平了道路。

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