State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Centre for Magnetic Resonance, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071, PR China.
J Proteome Res. 2010 Mar 5;9(3):1460-75. doi: 10.1021/pr900995m.
Plant metabonomic analysis is essential for understanding plant systems responses to osmotic stresses. To understand the comprehensive metabolic responses of Salvia miltiorrhiza Bunge (SMB) to continuous and exhaustive water depletion, we characterized the SMB metabonomic variations induced by three different drying processes using the combined NMR and LC-DAD-MS method. NMR results showed that SMB extracts were dominated by 29 primary metabolites such as sugars, carboxylic acids and amino acids, which were comprehensively reported for the first time, and 8 secondary metabolites including polyphenolic acids and diterpenoids. LC-DAD-MS methods detected 44 secondary metabolites, among which 5 polyphenolic acids together with genipin, umbelliferone and tormentic acid were found for the first time in this plant. We found that aqueous methanol was efficient in extracting both primary metabolites and polyphenolic acids, whereas chloroform-methanol was effective in selectively extracting diterpenoids. We further found that air- and sun-drying markedly affected both primary and secondary metabolisms of SMB by enhancing tanshinone and glutamate-mediated proline biosynthesis and altering carbohydrate and amino acid metabolisms. The shikimate-mediated biosynthesis of polyphenolic acids was promoted by air-drying but suppressed by sun-drying. These findings fill the gap of our understandings to the metabolic responses of S. miltiorrhiza Bunge to water depletion and demonstrated effectiveness of the combined NMR and LC-DAD-MS methods in plant metabonomic analysis.
植物代谢组学分析对于理解植物系统对渗透胁迫的反应至关重要。为了了解丹参（SMB）对连续和彻底缺水的综合代谢反应，我们采用 NMR 和 LC-DAD-MS 联合方法，对三种不同干燥过程诱导的 SMB 代谢组学变化进行了表征。NMR 结果表明，SMB 提取物主要由 29 种初级代谢物组成，如糖、羧酸和氨基酸，这是首次全面报道的，还有 8 种次级代谢物，包括多酚酸和二萜。LC-DAD-MS 方法检测到 44 种次级代谢物，其中 5 种多酚酸与京尼平、茴芹素和乌头酸一起，首次在该植物中发现。我们发现水甲醇有效地提取了初级代谢物和多酚酸，而氯仿-甲醇则有效地选择性提取了二萜。我们进一步发现，空气干燥和阳光干燥通过增强丹参酮和谷氨酸介导的脯氨酸生物合成以及改变碳水化合物和氨基酸代谢，显著影响 SMB 的初级和次级代谢。空气干燥促进了莽草酸介导的多酚酸生物合成，但阳光干燥则抑制了这一过程。这些发现填补了我们对丹参对水分亏缺代谢反应理解的空白，并证明了 NMR 和 LC-DAD-MS 联合方法在植物代谢组学分析中的有效性。
