Yang Da, Song Yeyang, Lu Anjin, Qin Lin, Tan Daopeng, Zhang Qianru, He Yuqi, Lu Yanliu
Key Lab of the Basic Pharmacology of the Ministry of Education, Zunyi Medical University, 6 West Xue-Fu Road, Zunyi 563009, China.
Guizhou Engineering Research Center of Industrial Key-technology for Dendrobium Nobile, Zunyi Medical University, 6 West Xue-Fu Road, Zunyi 563000, China.
Metabolites. 2023 Mar 6;13(3):389. doi: 10.3390/metabo13030389.
Kimura et Migo is a famous plant with a high medicinal value which has been recorded in the Chinese Pharmacopoeia (2020 Edition). The medicinal properties of are based on its chemical composition. However, there are no reports on how different cultivation methods affect its chemical composition. In order to reveal this issue, samples of the were collected in this study through tree epiphytic cultivation, stone epiphytic cultivation, and greenhouse cultivation. Polysaccharides were determined by phenol sulfuric acid method and secondary metabolites were detected by the UPLC-MS technique. In addition, with regards to metabolomics, we used multivariate analyses including principal component analysis (PCA) and orthogonal partial least squares analysis (OPLS-DA) to screen for differential metabolites which met the conditions of variable importance projection values >1, fold change >4, and < 0.05. The differential metabolites were taken further for metabolic pathway enrichment analysis, which was based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and validated by antioxidant activity. Comparing the three groups of samples according to the standards of the ChP (2020 edition), the results showed that the polysaccharide content of the samples from stony epiphytic cultivation and greenhouse cultivation was significantly higher than that of the samples from live tree epiphytic cultivation. Metabolomic analysis revealed that there were 185 differential metabolites among the 3 cultivation methods, with 99 of the differential metabolites being highest in the stone epiphytic cultivation. The results of the metabolic pathway enrichment analysis showed that the different cultivation strategies mainly effected four carbohydrate metabolic pathways, five secondary metabolite synthesis pathways, six amino acid metabolic pathways, one nucleotide metabolism pathway, three cofactor and vitamin metabolism pathways, and one translation pathway in genetic information processing. Furthermore, from stone epiphytic cultivation which had the best antioxidant activity was implicated in differential metabolite production. This study revealed the effects of different cultivation methods on the chemical composition of and also provided a reference for establishing the quality control standards to aid its development and utilization.
金粟兰是一种著名的具有高药用价值的植物，已被载入《中国药典》（2020年版）。其药用特性基于其化学成分。然而，关于不同栽培方法如何影响其化学成分尚无报道。为揭示这一问题，本研究通过树附生栽培、石附生栽培和温室栽培收集了金粟兰样本。采用苯酚硫酸法测定多糖，用超高效液相色谱-质谱联用技术检测次生代谢产物。此外，在代谢组学方面，我们使用了包括主成分分析（PCA）和正交偏最小二乘法分析（OPLS-DA）在内的多变量分析方法，以筛选出满足变量重要性投影值>1、变化倍数>4且P<0.05条件的差异代谢物。对差异代谢物进一步进行基于京都基因与基因组百科全书（KEGG）数据库的代谢途径富集分析，并通过抗氧化活性进行验证。按照《中国药典》（2020年版）标准比较三组样本，结果表明，石附生栽培和温室栽培样本的多糖含量显著高于树附生栽培样本。代谢组学分析显示，三种栽培方法中有185种差异代谢物，其中99种差异代谢物在石附生栽培中含量最高。代谢途径富集分析结果表明，不同栽培策略主要影响四条碳水化合物代谢途径、五条次生代谢物合成途径、六条氨基酸代谢途径、一条核苷酸代谢途径、三条辅因子和维生素代谢途径以及遗传信息处理中的一条翻译途径。此外，石附生栽培的金粟兰具有最佳抗氧化活性，这与差异代谢物的产生有关。本研究揭示了不同栽培方法对金粟兰化学成分的影响，也为建立质量控制标准以促进其开发利用提供了参考。
