| Hippocampus, which was first documented in the "Ben Cao Jing Ji Zhu", is a well-known traditional Chinese medicine. Five species of the genus Hippocampus were documented by Chinese Pharmacopoeia, and were listed in CITES Appendix II. The records about Hippocampus in the material medica of past dynasties were different. Due to the destruction of morphological characteristics after processing, the traditional identification of Chinese medicine on the basis of morphological characteristics is generally not accurate of difficult to ensure accuracy,Therefore it is very necessary to establish a rapid and accuratereliable method for the differentiation of the Hippocampus. Because it is rapid, accurate and convenient, DNA barcoding is used to identify Chinese medicinal materials. DNA barcoding for species identification is not influenced by the growth phase and morphology, which makes up for deficiencies of traditional morphological classification. Hippocampus were identified in this study using DNA barcoding on the basis of morphological studies. The study laid the foundation for the application of Hippocampus.The herbal literatures indicated that the earliest record for Hippocampus was in the "Ben Cao Jing Ji Zhu", and the ancient literatures.The literatures about morphological description of Hippocampus were few, and wereless could not provide a scientific basis for direct morphological identification of Hippocampus.The market survey of Hippocampus medicinal material in Chengdu lotus pond, Anhui Bozhou and Hebei Anguo was proceeded. The results showed that:1)compendia sales proportion of commercially available medicinal plants inlarger hippocampus, there are a total of10seahorse species were sold in medicine market and five commercially available varieties, are recorded in Chinese Pharmacopoeia the Pharmacopoeia of;2) There is a serious adulteration in medicine market and weight ratio of impurity to H.queenslandicus is46.58%;3)Sale standard for Hippocampus is complete external shape, smooth less prickly, big and clean surface, which is not standard without the strict classification sales,most of the hippocampus complete shape,smoothless prickly,a big,clean surface for the quality is better. On the basis of the market survey, we preliminary established the graphic retrieval table of Select Hippocampus medicine market surveys obtained the object of study lay the foundation for the rapid identification of Hippocampus medicinal materials.DNA barcoding showed thatthe15seahorse species that were identified based on morphological study were nine species, and the study for H.histrix is still in progress.The result suggested the necessity and importance using DNA barcoding to identify Hippocampus medicinal materials. Analysis of of COI gene of4bases in the seahorse breeds9commercially available:showed that A+T average content (59.4%)was much higher than the average G+C content (40.6%), which was consistent with the characteristics of vertebrate mitochondrial DNA. The results of genetic distance.The average intrespecific genetic distance (0.0853) was much greater than the average intraspecific genetic distance0.0033. The phylogenetic tree showed that nine seahorse species formed nine monophyletic, and the different individuals of the same species clusterred together with high bootstrap values. The results indicated that DNA barcoding could identify Hippocampus medicinal materials. In order to verify the feasibility of DNA barcodeing for commercial Hippocampus, we did a random verification experiment, which showed that DNA barcoding could accurately and quickly identify the random sample of Hippocampus. The study indicated that it was feasible using DNA barcoding technology to verify Hippocampus medicinal materials.In summary, mitchondrial COI gene as a standard DNA bacording for Hippocampus could rapidly and accuratly identifiy Hippocampus medicinal materials.The study improved and complemented the morphological study of traditional Chinese medicinal materials Hippocampus.
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