土壤因素对活血丹药材品质的影响机理研究

Soil is the natural medium of medicinal materials. The different physicochemical and biological characteristics of soil leads to the discrepancy of medicinal materials quality under different cultivated conditions. Glechoma longituba (Nakai) Kupr. (Labiatae) is an herbaceous medicinal plant. Its dried aerial part, Glechomae herba, is a standard medicinal material in the Chinese Pharmacopoeia and is used for the treatment of cholecystitis, gall-stone, kidney stone, etc. The wild population of G. longituba cannot meet growing need due to the outstanding imbalance between supply and demand. Based on our previous work, populations of G. longituba showed remarkable differences in chemical composition depending on the provenance of plants which is related with environmental and soil factor. However, the main environmental factor, especially soil factor, influencing Glechomae herba quality remains poorly understood. In this project, we aim to collect G. longituba plants and related soil from different locations, to explore the relationship between the Glechomae herba quality formation and soil factors from different locations on the basis of and analysis methods of medicinal materials ingredient analysis and soil physico-chemical properties, to investigate the influence and mechanism of soil factor on the quality of Glechomae herba through single-factor experiment, and to determine the definitive soil factor for high quality G. longituba formation. Based on above-mentioned works, our objective is to establish scientific basis and technical support for the determination of appropriate planting base and standardized production of G. longituba.

土壤作为药材的"天然培养基"，由其理化和生物性质构成的土壤生物作用造成了不同栽培条件下中药材质量的显著差异。活血丹G. longituba (Nakai) Kupr.为唇形科多年生草本植物，干燥地上部分入药，名连钱草，主治胆囊炎、肾结石、膀胱结石等症，市场需求量逐年增大，供不应求。前期研究表明，不同地理种源活血丹有效成分含量差异显著，与环境和土壤性质有关，但控制活血丹药材品质的环境和土壤的具体主控因素不清楚。本研究将采集不同地理种源活血丹植株及土壤样品，借助中药成分分析和土壤理化分析技术，通过数理统计方法，初步筛选影响活血丹药材品质的土壤主控因子；采用单因素试验逐一验证土壤主控因子及其对活血丹品质的影响机理；通过主控因子的多因素综合作用研究，明确优质活血丹生长的最佳土壤条件，以期为活血丹适宜生产区的合理分区和定向培育提供理论依据和技术支持。

活血丹G. longituba (Nakai) Kupr.为唇形科多年生草本植物，其干燥地上部分入药，《中国药典》收录为“连钱草”。不同地理种源活血丹的活性成分含量差异较大，土壤作为植物的“天然培养基”，其理化和生物性质构成的土壤-生物作用与中药材质量关系密切。为探寻高品质活血丹形成的土壤条件，本项目开展了研究工作，目前取得如下主要结果：建立了活血丹内酚酸类（绿原酸、咖啡酸和迷迭香酸）的超声提取条件（40%乙醇溶液、液料比40 mL·g-1、60ºC水浴中超声提取100 min）与高效液相测定方法，优化了原有的活血丹三萜酸类（熊果酸与齐墩果酸）的提取方法与高效液相测定方法和总黄酮检测方法。系统收集活血丹分布区内不同地理种源活血丹及根际土壤并引种后，根据活血丹药材品质与根际土壤理化性质和微生物种类等差异、变化，利用灰色关联度分析与偏最小二乘法（PLS）筛选出了影响活血丹药材品质形成的土壤主要因子——土壤酸碱度、土壤容重、有机质含量、磷和铁。分别研究了其对活血丹产量和质量形成的影响，各因子对活血丹生长及其药材品质的促进均存在阈值，利于活血丹产量形成和活性成分积累的各因子适宜范围分别：土壤pH6.5±1、土壤容重1.0±0.1g·cm-3、土壤有机质含量71.5±18.3 g·kg-1、磷的中浓度处理（1.0 mmol·L-1）和铁的低浓度处理(15 μmol·L-1)。最后利用正交试验确定高品质活血丹的土壤条件为：土壤pH6.5、土壤容重1.0 g·cm-3、土壤有机质含量89.8 g·kg-1；最佳土壤磷铁组合为额外施加0.9 g·kg-1的磷素和0.12 g·kg-1的铁素，其次施加0.9 g·kg-1磷素和不施加铁素的组合及施加0.9 g·kg-1磷素和0.24 g·kg-1铁素的组合亦较有利于活血丹药材品质的提高。研究结果对于高品质中药材定向培育和适宜生产区的合理分区可以提供理论依据和技术支持。