title: Identification of ungulates used in a traditional Chinese medicine with DNA barcoding technology.
abstract: Horns of Saiga antelope (Saiga tatarica) have always been an ingredient of "Lingyangjiao", a traditional Chinese medicine (TCM). Persistent hunting for Saiga antelope has already threatened the survival of critical endangered populations in wild. To control the growing pressure, CITES and Chinese government have legislated for monitoring the trade of Saiga horns. However, similar ungulate horns are difficult to identify by their morphological characteristics, which has impeded the law enforcement. Besides Saiga antelope, other seven ungulate species which have similar horns are also sold and marked as "Lingyangjiao" in TCM markets to offset shortage of Saiga antelope horns. Such species are Gazella subgutturosa, Pantholops hodgsonii, Procapra picticaudata, Procapra gutturosa, Procapra przewalskii, Capra hircus, and Ovis aries. Our study aimed at implementing DNA barcoding technology to diagnose Saiga horns and the substitutes. We successfully extracted genomic DNA from horn samples. We recovered COI sequences of 644 bp with specific primers and 349 bp with nested PCR primers designed for degraded horn samples. The mean interspecific genetic distance of data set of the 644-bp full barcodes and the 349-bp mini-barcodes was 14.96% and 15.38%, respectively, and the mean intraspecific distance was 0.24% and 0.20%, respectively. Each species formed independent clades in neighbor-joining (NJ) phylogenetic tree of the two data sets with >99% supporting values, except P. gutturosa and P. przewalskii. The deep genetic distances gap and clear species clades in NJ tree of either full barcodes or mini-barcodes suggest that barcoding technology is an effective tool to diagnose Saiga horns and their substitutes. Barcoding diagnosis protocol developed here will simplify diagnosis of "Lingyangjiao" species and will facilitate conservation of endangered ungulates involved in TCM "Lingyangjiao" markets, especially the Saiga antelope.
authors.affiliation: Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China ; University of Chinese Academy of Sciences No. 19 (A) Yuquan Road, Beijing, 100049, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China ; University of Chinese Academy of Sciences No. 19 (A) Yuquan Road, Beijing, 100049, China ; Endangered Species Scientific Commission of the People's Republic of China Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: School of Resources and Environmental Engineering, Anhui University No. 111 Jiu Long Road, Hefei, 230601, Anhui Province, China.
authors.forename: Chunlin
authors.initials: C
authors.lastname: Li
authors.affiliation: Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China.
authors.forename: Xiaoge
authors.initials: X
authors.lastname: Ping
authors.affiliation: Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China ; University of Chinese Academy of Sciences No. 19 (A) Yuquan Road, Beijing, 100049, China.
authors.forename: Shaopeng
authors.initials: S
authors.lastname: Cui
authors.affiliation: Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences Chaoyang District, No. 1 Beichen West Road, Beijing, 100101, China.
authors.forename: Songhua
authors.initials: S
authors.lastname: Tang
authors.affiliation: Wildlife Conservation Office of Altay Prefecture 836500, Altay, Xinjiang, China.
authors.forename: Hongjun
authors.initials: H
authors.lastname: Chu
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, No. 26 Hexing Road Xiangfang District Harbin, 150040, Heilongjiang Province, China.
authors.forename: Binwan
authors.initials: B
authors.lastname: Liu
keywords: Conservation genetics; DNA barcode; DNA identification; illegal trade; traditional Chinese medicines; wildlife forensics