Identification of Saigae Tataricae Cornu in Chinese materia medica preparation using DNA barcoding / 中草药

ABSTRACT

Objective: To discuss the feasibility of using partial sequence of mitochondrial cytochrome C oxidase subunit I (COI) gene as DNA barcoding to identify Saigae Tataricae Cornu (STC) in Chinese materia medica (CMM) preparation. Methods: The DNA barcoding identification was constructed in STC samples from eight different manufactories. The genome DNA was extracted by DNA extraction kit from adequately ground samples. The COI nucleotide sequences were PCR amplified with the Primers LCO1490 and HC02198 or specified primer 0703 and sequenced bi-directionally. The BLAST comparison was made in GeneBank, and the phylogenetic trees were constructed with the homologisation tree (DNAMAN) and Neighbor-Joining (NJ, MEGA 5.2) method. Saiga tatarica (gb|JN632700.1) and STC were selected as reference medicinal materials and the genetic distance was calculated using Kimura-2-parameter. Results: The lengths of partial mitochondrial COI gene collected from STC in eight samples of CMM preparation were about 658 bp. The phylogenetic tree showed that the samples of CMM preparation and reference substance assembled distinctly, and were completely separated from the outgroup. The intraspecific genetic distances of these samples ranged in 0-0.064 with an average of 0.020, and the interspecific genetic distances ranged in 0.167-0.195. Conclusion: The mitochondrial COI gene is a valid DNA barcoding gene for STC identification in CMM preparation. But other organs in antelope also contain the same gene. In order to ensure the accuracy of the results, other texst methods also need to be used cooperatively.