Xu Yue, Cao Yingping, Tie Fangfang, Kong Xiuya, Liu Yuchen, Zhang Yaru, Guan Wenna, Hu Na, Wang Honglun, Qin Xiaochun, Wu Zhenying, Fu Chunxiang
School of Chemistry and Chemical Engineering, School of Biological Science and Technology, University of Jinan, Jinan, China.
Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.
Plant Biotechnol J. 2025 May;23(5):1713-1724. doi: 10.1111/pbi.70001. Epub 2025 Feb 18.
The snow lotus species Saussurea involucrata (Kar. & Kir.) Sch.Bip., an endangered traditional Chinese herb, belongs to a genus of the Asteraceae family. Syringin present in S. involucrata stands as one of the predominant bioactive compounds. However, the biosynthetic pathway of syringin remains largely elusive. Here, S. involucrata suspension cell culture was subjected to methyl jasmonate (MeJA) treatment, which stimulated the synthesis of syringin, increasing its content by up to 3.9-fold. Comparative transcriptome analysis revealed that genes involved in syringin biosynthesis were generally upregulated in response to MeJA. Furthermore, two candidate UDP-glycosyltransferase genes, SiUGT72BZ2 and SiUGT72CY1, were identified through phylogenetic tree and expression profiling analyses. Overexpression of SiUGT72BZ2 (BZ2_OE) and SiUGT72CY1 (CY1_OE) in S. involucrata suspension cell cultures led to 15.2- and 5.9-fold higher syringin levels than empty vector control cultures, respectively. Notably, upregulation of SiUGT72BZ2 enhanced the biosynthesis of coniferin as well. In contrast, only trace amounts of coniferin were present in control and CY1_OE cell cultures. Subsequent anti-inflammatory assays using lipopolysaccharide (LPS)-stimulated RAW264.7 cells demonstrated that the extracts from these cell cultures possessed remarkable anti-inflammatory properties. Most strikingly, the BZ2_OE cultures exhibited superior anti-inflammatory effects compared to the control and CY1_OE. In conclusion, our research has not only identified the key enzymes in syringin synthesis but also, through genetic engineering, has generated novel cell culture resources enriched with syringin and coniferin, and enhanced anti-inflammatory activities, highlighting the potential of S. involucrata cell culture as an alternative for wild snow lotus resources.
雪莲花物种天山雪莲（Saussurea involucrata (Kar. & Kir.) Sch.Bip.）是一种濒危的传统中药材，属于菊科的一个属。天山雪莲中存在的紫丁香苷是主要的生物活性化合物之一。然而，紫丁香苷的生物合成途径在很大程度上仍然不清楚。在这里，天山雪莲悬浮细胞培养物用茉莉酸甲酯（MeJA）处理，这刺激了紫丁香苷的合成，其含量增加了3.9倍。比较转录组分析表明，参与紫丁香苷生物合成的基因通常在MeJA处理下上调。此外，通过系统发育树和表达谱分析鉴定了两个候选UDP-糖基转移酶基因SiUGT72BZ2和SiUGT72CY1。在天山雪莲悬浮细胞培养物中过表达SiUGT72BZ2（BZ2_OE）和SiUGT72CY1（CY1_OE），导致紫丁香苷水平分别比空载体对照培养物高15.2倍和5.9倍。值得注意的是，SiUGT72BZ2的上调也增强了松柏苷元的生物合成。相比之下，对照和CY1_OE细胞培养物中仅存在痕量的松柏苷元。随后使用脂多糖（LPS）刺激的RAW264.7细胞进行的抗炎试验表明，这些细胞培养物的提取物具有显著的抗炎特性。最引人注目的是，与对照和CY1_OE相比，BZ2_OE培养物表现出更强的抗炎作用。总之，我们的研究不仅鉴定了紫丁香苷合成中的关键酶，而且通过基因工程产生了富含紫丁香苷和松柏苷元且具有增强抗炎活性的新型细胞培养资源，突出了天山雪莲细胞培养作为野生雪莲资源替代品的潜力。
