Key Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, Zhejiang, China.
Tianjin Tasly Holding Group Co., Ltd., Tianjin 300410, China.
Gene. 2024 Mar 20;899:148094. doi: 10.1016/j.gene.2023.148094. Epub 2023 Dec 22.
Salvia miltiorrhiza, a prominent traditional Chinese medicinal resource, has been extensively employed in the management of cardiovascular and cerebrovascular ailments. Ensuring the consistency of S. miltiorrhiza raw materials revolves around the imperative task of maintaining stable tanshinones content and composition. An effective approach in this regard involves the utilization of endophytic fungi as inducers. Within this context, our study spotlights an endophytic fungus, Penicillium steckii DF33, isolated from the roots of S. miltiorrhiza. Remarkably, this fungus has demonstrated a significant capacity to boost the biosynthesis and accumulation of tanshinones. The primary objective of this investigation is to elucidate the underlying regulatory mechanism by which DF33 enhances and regulates the biosynthesis and accumulation of tanshinones. This is achieved through its influence on the differential expression of crucial CYP450 genes within the S. miltiorrhiza hairy roots system. The results revealed that the DF33 elicitor not only promotes the growth of hairy roots but also enhances the accumulation of tanshinones. Notably, the content of cryptotanshinone was reached 1.6452 ± 0.0925 mg g, a fourfold increase compared to the control group. Our qRT-PCR results further demonstrate that the DF33 elicitor significantly up-regulates the expression of most key enzyme genes (GGPPS, CPS1, KSL1, CYP76AH1, CYP76AH3, CYP76AK1, CYP71D411) involved in the tanshinone biosynthesis pathway. This effect is particularly pronounced in certain critical CYP450 genes and Tanshinone ⅡA synthase (SmTⅡAS), with their expression levels peaking at 7 days or 14 days, respectively. In summary, endophytic P. steckii DF33 primarily enhances tanshinone biosynthesis by elevating the expression levels of pivotal enzyme genes associated with the modification and transformation stages within the tanshinone biosynthesis pathway. These findings underscore the potential of employing plant probiotics, specifically endophytic and root-associated microbes, to facilitate the biosynthesis and transformation of vital constituents in medicinal plants, and this approach holds promise for enhancing the quality of traditional Chinese medicinal materials.
丹参，一种重要的传统中药资源，广泛应用于心脑血管疾病的治疗。确保丹参原料的一致性，关键在于维持丹参酮含量和组成的稳定。在这方面，一种有效的方法是利用内生真菌作为诱导剂。在这方面，我们的研究重点是一种内生真菌，即从丹参根部分离得到的青霉 steckii DF33。值得注意的是，这种真菌具有显著提高丹参酮生物合成和积累的能力。本研究的主要目的是阐明 DF33 增强和调节丹参酮生物合成和积累的潜在调控机制。这是通过其对丹参毛状根系统中关键 CYP450 基因的差异表达的影响来实现的。结果表明，DF33 诱导子不仅促进了毛状根的生长，而且还增强了丹参酮的积累。值得注意的是，隐丹参酮的含量达到 1.6452±0.0925mg/g，比对照组增加了四倍。我们的 qRT-PCR 结果进一步表明，DF33 诱导子显著上调了大多数关键酶基因（GGPPS、CPS1、KSL1、CYP76AH1、CYP76AH3、CYP76AK1、CYP71D411）的表达，这些基因参与了丹参酮的生物合成途径。这种作用在某些关键的 CYP450 基因和丹参酮 ⅡA 合酶（SmTⅡAS）中尤为明显，它们的表达水平分别在 7 天或 14 天达到峰值。总之，内生青霉 steckii DF33 主要通过提高丹参酮生物合成途径中修饰和转化阶段相关关键酶基因的表达水平来增强丹参酮的生物合成。这些发现强调了利用植物益生菌，特别是内生菌和根际微生物，促进药用植物中重要成分的生物合成和转化的潜力，这一方法有望提高传统中药材料的质量。
