Detection the eDNA of Batrachuperus taibaiensis from the Zhouzhi Heihe River Using a Nested PCR Method and DNA Barcoding

The Taibai stream salamander ( ) is a recently described species of the genus that occurs in the Zhouzhi Heihe River and is endangered in its native range. Here, we have established a method for water environmental DNA (eDNA) analysis of using a series of optimizations. We have designed a specific s...

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Bibliographic Details
Published in:Animals (Basel) 2022-04, Vol.12 (9), p.1105
Main Authors: Ma, Hongying, Zhang, Han, Deng, Jie, Zhao, Hu, Kong, Fei, Jiang, Wei, Zhang, Hongxing, Dong, Xianggui, Wang, Qijun
Format: Article
Language:English
Subjects:
Alcohol
Batarchuperus taibaiensis
Batrachuperus
Batrachuperus taibaiensis
Biodiversity
Deoxyribonucleic acid
DNA
DNA barcoding
eDNA
Endemic species
Environmental DNA
genus-specific primer
Methods
nested PCR
Nucleotide sequence
Polymerase chain reaction
Protected species
Rare species
Rivers
Sequence analysis
Sequences
Water analysis
Water quality
Water sampling
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Summary:The Taibai stream salamander ( ) is a recently described species of the genus that occurs in the Zhouzhi Heihe River and is endangered in its native range. Here, we have established a method for water environmental DNA (eDNA) analysis of using a series of optimizations. We have designed a specific set of primers for the genus to amplify a 160 bp fragment of . The sequences were obtained from nested PCR on eDNA extracted from water samples, after which DNA barcoding was performed according to sequence analysis to determine the presence of the target species in the water. The method was validated using water from the Zhouzhi Heihe River with known populations and found that eDNA can move at least 150 m downstream from its point of origin. This study is the first to establish an optimal method for obtaining the eDNA of from water samples, which provides a theoretical basis for resource investigation and the protection of in future research. It is also an example of the eDNA extraction of other species that live in similar waters and are less genetically diverse between species.
ISSN:2076-2615
2076-2615
DOI:10.3390/ani12091105