Biodiv Sci ›› 2023, Vol. 31 ›› Issue (7): 22661.  DOI: 10.17520/biods.2022661

• Original Papers: Animal Diversity • Previous Articles     Next Articles

The establishment of terrestrial vertebrate genetic resource bank and species identification based on DNA barcoding in Wanglang National Nature Reserve

Chao Xing1,2, Yi Lin2,3, Zhiqiang Zhou4, Lianjun Zhao4, Shiwei Jiang5, Zhenzhen Lin2,*(), Jiliang Xu1,*(), Xiangjiang Zhan2   

  1. 1. College of Ecology and Nature Conservation, Beijing Forestry University, Beijing 100083
    2. Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101
    3. University of Chinese Academy of Sciences, Beijing 100049
    4. Wanglang National Nature Reserve Administration, Pingwu, Sichuan 622550
    5. Pingwu Forestry and Grassland Bureau, Pingwu, Sichuan 622550
  • Received:2022-11-25 Accepted:2023-04-14 Online:2023-07-20 Published:2023-04-25
  • Contact: *E-mail:;


Aims: DNA barcoding using the short, highly conserved regions of the genome to make species-level identifications, is widely used in the species identification and diversity assessment of plants and animals in biodiversity conservation research. Wanglang National Nature Reserve is a global biodiversity hot spot located in the eastern part of Qinghai-Tibet Plateau with high level of biodiversity and has a leading role in the wildlife conservation in China. Although a lot of survey data of species diversity have been collected, genetic resource of terrestrial vertebrates in the reserve remains largely unclear. To address this, our study aims to establish a DNA barcoding database for terrestrial vertebrate species for the reserve.

Methods: We collected genetic samples mainly using non-invasive sampling along the line transects in Wanglang National Nature Reserve and from museum specimens. We extracted DNA from the samples to obtain the barcode sequences by PCR. Each DNA barcode sequences were blasted against NCBI database for species identification. For those sequences that we could not directly get the matches in the reference database, we constructed phylogenetic trees using MEGA 11.0 with the aim to identify the species based on the sequences of its relative species from the public database.

Results: The total of 314 samples including tissues from carcasses, feathers, feces and eggshells were collected and 74 species were identified using 16 pairs DNA barcoding primers. The sequences of mitochondrial COI, Cytb, 12S-16S, 16S genes and sequences of nuclear RAG1 gene of Scutiger wanglangensis were firstly reported. Compared with other wildlife monitoring techniques such as camera traps, drift fences and pitfall traps in the same time interval, DNA barcoding is more efficient in all taxon groups and has better performance in bird monitoring. Finally, A terrestrial vertebrate genetic resource bank contains 314 substantial samples and 216 DNA barcodes has been established.

Conclusion: Our study established an elementary DNA barcoding database of 74 terrestrial vertebrate species in the reserve, highlights the utility of DNA barcoding technique in wild animal diversity surveys, and provides a basic catalog for assessing local species diversity to improve conservation research and management in the reserve.

Key words: DNA barcoding, Wanglang National Nature Reserve, terrestrial vertebrates, biodiversity, barcode database