天麻熄风胶囊的DNA条形码鉴定研究

Studies for DNA Barcoding Identification of the Tianma Xifeng Capsule

目的:探讨天麻熄风胶囊的DNA条形码鉴定方法与价值。方法:2015年1月到11月,选择3批天麻熄风胶囊(批号20150918、200150919、20151019)进行显微鉴定与DNA条形码鉴定,选择的基因序列为天麻素线粒体A基因,进行基因测序与遗传分析。结果:显微鉴定显示天麻细胞无色,呈现长卵形、长椭圆形或类圆形,遇碘液显棕色;蔓荆子非腺毛2-3个细胞,顶端细胞基部稍粗;甘草纤维成束、壁厚,周围薄壁细胞可形成晶纤维;半夏椭圆形细胞存在于粘液细胞中。3批天麻熄风胶囊的天麻素线粒体A基因特定序列中,A含量为27.9%、C为24.9%、G为18.3%、T为28.9%,G-T含量明显高于G/C含量(43.2%)(P<0.05)。遗传分析显示20150918批次的种间最小K-2P距离为0.006,种间最大K-2P距离为0.241;20150919批次种间最小K-2P距离为0.008,最大K-2P距离为0.074;20151019批次的种间最小K-2P距离为0.008,种间最大K-2P距离为0.133。结论:天麻熄风胶囊成分复杂,显微鉴定技术存在一定的缺陷,DNA条形码鉴定能有效反映胶囊成分,且能很好判定药物批次的种内与种间变异,具有很好的鉴别价值。

Objective： To investigate the DNA barcoding identification ways and effects for the Tianma Xifeng Capsule. Methods： From January 2015 to November 2015, selected 3 batches （batch 20150918, 200150919 and 20151019） of Tianma Xifeng capsules for microscopic identification and DNA barcoding, the gene sequences were selected with gastrodin mitochondria A gene, and were given the gene sequencing and genetic analysis. Results： Microscopic identification displayed that the Tianma cell were colorless and showed long oval, oblong or round, in case of iodine it showed brown; Non-glandular hairs had 2-3 cells for viticis, and the apical cell base was thicker; Banxia Oval cells were present in the mucus cells; licorice fiber was bundles, wall thickness, and the surrounding parenchyma cells can form crystal fibers, the average contents of A gene for the specific sequence A, C, G, T, in the 3 batches of capsules Xifeng Tianma were 27.9%, 24.9%, 18.3%, to 28.9%, the G-T contents （56.8%） were significantly higher than the G/C content （43.2%） （P 〈0. 05）. Genetic analysis revealed that the smallest K-2P distance in the 20150918 batch was 0.006, the maximum K-2P distance was 0.241; and for the batches of 200150919 and 20151019 were 0.008, 0.074 and 0.008, 0.133, respectively. Conclusion： The composition for Tianma Xifeng capsule is complex, and the identification for it with microscopic technology has some flaws, while the DNA barcoding has a good discriminating value since it can effectively reflect the capsule ingredient, and determine within the good batch of drugs and variation among species.