Identification of 24 Rana Species Including Rana dybowskii and Rana chensinensis Based on COⅠ Sequences
10.13422/j.cnki.syfjx.20211015
- VernacularTitle:基于COⅠ基因对东北林蛙、中国林蛙等蛙属24种的鉴定
- Author:
Meng-hu WANG
1
;
Yi-fan SUN
2
;
Liang XU
3
;
Ting-guo KANG
3
;
Teng-teng ZHANG
1
;
Ya-feng ZUO
1
;
Li-ting ZHU
3
;
Xiang-song MENG
1
;
Jian TANG
1
;
Qian XU
4
Author Information
1. Bozhou University，Bozhou 236800，China
2. Bozhou Vocational and Technical College， Bozhou 236800，China
3. Liaoning University of Traditional Chinese Medicine，Dalian 116600，China
4. The Second Affiliated Hospital of Anhui University of Chinese Medicine，Hefei 230061，China
- Publication Type:Research Article
- Keywords:
Rana dybowskii;
R. chensinensis;
Ranidae;
Rana;
DNA barcoding;
mitochondrial cytochrome C oxidase subunit Ⅰ （COⅠ） gene
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(16):150-158
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To identify 24 Rana species such as Rana dybowskii by mitochondrial cytochrome C oxidase subunit I （COⅠ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of Rana species as well as the discovery of new species. Method:R. dybowskii， R. chensinensis， R. amurensis， R. culaiensis， and R. huanrenesis， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification and sequencing. A total of 50 COⅠ gene sequences were obtained. Then 163 COⅠ gene sequences for 24 species of Rana and one COⅠ gene sequence for Pelophylax， Odorrana， Nidirana， Hylarana， and Amolops were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of COⅠ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The COⅠ gene sequences of 24 Rana species including R. dybowskii were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between R. sangzhiensis and R. zhengi was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. R. sangzhiensis and R. zhengi were clustered into one branch and some R. dybowskii and R. uenoi into one branch. There were two separate branches for R. chensinensis and the other species were all clustered independently. Conclusion:COⅠ-based DNA barcoding enabled the identification of 24 species of Rana including R.dybowskii. The findings supported that R. sangzhiensis， R. zhengi， R. coreana， and R. kunyuensis were the same species. One branch of R. chensinensis might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that COⅠ-based DNA barcoding allows not only the identification of 24 species of Rana including R. dybowskii but also the classification of ranidae species and the discovery of new species or subspecies.
