Xu H, Fabricant D S, Piersen C E, Bolton J L, Pezzuto J M, Fong H, Totura S, Farnsworth N R, Constantinou A I
UIC/NIH Center for Botanical Dietary Supplements Research, University of Illinois at Chicago, Chicago, Illinois, USA.
Phytomedicine. 2002 Dec;9(8):757-62. doi: 10.1078/094471102321621403.
Traditional taxonomic methods of botanical identification that rely primarily on morphological observations cannot be used efficiently when only powdered plant materials are available. Thus, our objectives were to determine if we could apply a molecular approach to: a) produce unique DNA profiles that are characteristic of the species, and b) determine if the geographical area or time of collection influences these DNA profiles. Towards this end, random amplified polymorphic DNA (RAPD) analyses were performed on a number of botanicals currently used for women's health. The test materials included samples from three species each of the genera Cimicifuga (Actaea) and Trifolium, as well as samples of Vitex agnus-castus L., Glycyrrhiza glabra L., Gingko biloba L., Valeriana officinalis L., Angelica sinensis (Oliv.) Diels, Viburnum prunifolium L., Humulus lupulus L., Vaccinium macrocarpon Ait., Panax ginseng C.A. Mey. Cimicifuga racemosa (L.) Nutt. and Trifolium pratense L. are currently under clinical investigation in our basic research laboratories and medical clinic for the relief of post-menopausal symptoms. Characteristic profiles produced with the OPC-15 primer could distinguish the three Cimicifuga species: C. racemosa, C. americana and C. rubifolia. Similar results were obtained with the three Trifolium species: Trifolium pratense L., Trifolium incarnatum L., and Trifolium repens L. Accessions of cultivated T. pratense collected from the same field at different times, produced identical profiles. Accessions of Cimicifuga species collected from different geographical areas produced similar but not identical DNA profiles; however, species-specific DNA fragments were identified. These results demonstrate that RAPD analysis can be applied to distinguish species when only powdered material is available for testing. This methodology can be applied to identify species of commercial value regardless of collection time or geographic area.
主要依靠形态学观察的传统植物分类鉴定方法，在仅能获得植物粉末材料时无法有效使用。因此，我们的目标是确定是否可以应用分子方法来：a）生成该物种特有的独特DNA图谱，以及b）确定采集的地理区域或时间是否会影响这些DNA图谱。为此，我们对目前用于女性健康的多种植物进行了随机扩增多态性DNA（RAPD）分析。测试材料包括升麻属（类叶升麻属）和三叶草属各三个物种的样本，以及穗花牡荆、光果甘草、银杏、缬草、当归、李叶荚蒾、啤酒花、大果越橘、人参的样本。总状升麻和红车轴草目前正在我们的基础研究实验室和诊所进行临床研究，用于缓解绝经后症状。用OPC - 15引物产生的特征图谱可以区分三种升麻属物种：总状升麻、美洲升麻和齿叶升麻。三种三叶草属物种：红车轴草、绛车轴草和白车轴草也得到了类似结果。在不同时间从同一田地采集的栽培红车轴草样本产生了相同的图谱。从不同地理区域采集的升麻属物种样本产生了相似但不完全相同的DNA图谱；然而，鉴定出了物种特异性的DNA片段。这些结果表明，当仅能获得粉末状材料用于测试时，RAPD分析可用于区分物种。这种方法可用于鉴定具有商业价值的物种，而不论采集时间或地理区域如何。
