虎杖的资源、品质与药效的相关性研究

【作者】 马云桐；

【导师】 万德光；

【作者基本信息】 成都中医药大学 ， 中药学， 2006， 博士

【摘要】 中药虎杖为蓼科植物虎杖Polygonum cuspidatum Sieb．et Zucc．根及根茎，在我国23个省、市有分布，但汾州并非文献记载的地道产地，近代虎杖以西南、陕南等地为其主产区。对虎杖资源的调查表明，资源正在遭受较为严重的破坏。为实现虎杖资源的可持续利用，应对资源进行分区优化配置，重点发展以西南、陕南为中心的虎杖优质药材商品基地。 本文对虎杖不同产地、不同季节、不同部位的有效成分含量的比较，研究结果表明有效成份的含量韧皮部高于木质部，成分的积累以9月为高峰期。部分产地及商品药材中大黄素的含量较《中国药典》规定标准略低，而虎杖苷的含量高出10～15倍。对虎杖的重金属限量考察结果符合国家限量标准。并建立了虎杖药材的HPLC指纹图谱，各产地的不同药材相似度较高。 首次综合运用RAPD、ISSR、SRAP标记技术构建了虎杖的DNA指纹图谱；虎杖的种质资源具有较为丰富的遗传多样性，22个引物共得到98条扩增DNA片段，扩增片段90.8％具有多态性，其中有17个引物(77.3％)扩增产物具多态性。通过聚类分析表明：虎杖的种质遗传多样性同地理分布有一定的相关性。首次对虎杖内生真菌进行了研究，虎杖内生真菌种群在类群与数量上具有多样性，两种成分的积累与拟青霉菌属、地霉菌属分布具有拮抗性的趋势，其原因有待进一步研究。 首次对虎杖生长的不同环境因子与虎杖中大黄素、虎杖苷的相关性进行了研究，氮和有机质是影响大黄素与虎杖苷含量的共同因素，K肥有利于大黄素含量的提高。对气候因子的分析，气温、日照因子对虎杖苷与大黄素的积累起主导作用，气温对虎杖苷有正向促进作用，日照时效对虎杖苷与大黄素有制约作用。Fe、Mg、K矿质元素有利于大黄素的积累，Al有利于虎杖苷的形成；“Fe、Al协同”因子和“促光合矿质”因子对虎杖中大黄素、虎杖苷含量增加贡献率高，植株体内Fe、Al、Ca呈同步富集的趋势；而Mg的富集，Mn、Zn、Cu的排泌可促进大黄素的合成，K富集使虎杖苷的含量下降。因此，在选择种植基地时，应在荫蔽、温暖的环境，加强对土壤Zn、Ca、Fe、Mg、K、Al的人为调控，种植中应注意补充速效性的N、P、K肥料三要素，以确保虎杖的质量。 首次建立了虎杖品质与药效相关性的数学模型，该模型较好的反应与印证了前面各章所做研究与分析的结果。模型的预测值与实测值十分相近，这对虎杖的种植基地的选择具有重要的指导意义。更多还原

【Abstract】 Huzhang, Rhizoma et Radix Polygoni Cuspidati, was widely used in China for a long time. According to the investigation on the resources of Polygonum cuspidatum, it was discovered that the Polygonaceae plant, Polygonum cuspidatum Sieb. Et Zucc. distributed in 23 provinces in China totally. However, the resources had been severely damaged in these places by degrees. The original productive place was not in Fenzhou as ancient documents recorded, but was in Southwest China and south Shanxi province. Therefore, the distribution of the resources should be districtly optimized for continuous utilization. We suggest to set up some planting bases of Huzhangi with high quality on these areas.After researching on the active components of Huzhang in different productive areasand different seasons, we found that the contents active components in phloem was higher than in xylem. The active components accumulated most in September in plants. Compared with the standard of Huzhang in Chinese Pharmacopoeia, the content of emodin in Huzhang from part of the production areas or from market was slightly decreased, while the content of polydatin was significantly 10-15 times more than that of the standard. Meanwhile, contents of heavy metals in Huzhang accorded with limitation standard as Chinese administration declared. As a good result, it will benefit for exportion of resveratol, which was extracted from Polygonum cuspidatum. The HPLC fingerprints of Polygonum cuspidatum was established for the first time. Results indicated that the HPLC fingerprints of Huzhang from different productive places were similar.RAPD, ISSR, SRAP marker techniques were applicated to study the genetic diversity of germplasm resources of Polygonum cuspidatum and its DNA fingerprints was established. With RAPD, ISSR, SRAP marker, the DNA fingerprints of Ploygonum cuspidatum were established for the first time. The germplasm resources of Polygonum cuspidatum has a rich genetic diversity. We harvested 98 DNA fragments with 22 primers. 90.8% of the DNA fragments had genetic diversity, while products from 17 primers of the total 22 primers had genetic diversity. Based on cluster analyzing, we found that the genetic diversity of germplasm resources of Polygonum cuspidatum were related to its geographic distribution. For the first time the endogeny fungal populations in Polygonum cuspidatum were studied. Results show that the fungals taxa and quantity showed varieties. The accumulation of two components更多还原