DNA barcoding of Artiodactyla in cases involved in wildlife for species identification

doi:10.3969/j.issn.1000-2006.202004013

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (6): 27-32.doi: 10.3969/j.issn.1000-2006.202004013

Special Issue: 野生动物保护与疫病防控专题

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DNA barcoding of Artiodactyla in cases involved in wildlife for species identification

LIU Dawei(), HOU Senlin^*(), ZHOU Yongwu, FEI Yiling, HE Jian

Key Laboratory of Wildlife Evidence Technology, Forest Police Forensic Identification Center, National Forestry and Grassland Administration,Nanjing Forest Police College, Nanjing 210023, China

Received:2020-04-08 Revised:2020-08-28 Online:2020-11-30 Published:2020-12-07
Contact: HOU Senlin E-mail:dwliu@nfpc.edu.cn;senlin_hou@163.com

Abstract

Abstract:

【Objective】In this study, we aimed to identify Artiodactyla species in cases of wildlife destruction, using mitochondrial DNA barcoding. 【Method】COI and Cyt b sequences in 67 samples from 16 species were amplified. Animal samples were identified using Blast search on the GenBank sequences. Intra-and inter-specific genetic distances were compared, and phylogenetic trees were constructed using MEGA 7.0. 【Result】In a homology analysis, the Cyt b sequences of Capreolus pygargus and Capreolus capreouls were 99.64%-99.82% homologous; this sequence could not distinguish two closely related species, whereas other COI and Cyt b sequences could. The intra-specific genetic distance was 0-1.4% for COI and 0-0.5% for Cyt b, whereas the inter-specific genetic distance was 1.8%-18.8% for COI and 4.0%-15.8% for Cyt b. An obvious barcoding gap was detected in both markers for all species. The phylogenetic tree showed that samples from the same species formed monophyletic groups with high support values. The gaps between different species were clear, and all samples from each species formed a single branch with a high support value.【Conclusion】COI and Cyt b may serve as molecular markers for Artiodactyla identification in cases involving wildlife resources. To minimize the risk of false identification, the two DNA barcodes could be used simultaneously, especially for the closely related species.

Key words: Artiodactyla, DNA barcoding, species identification, COI, Cyt b, gene fragment

CLC Number:

Q34
S718.6

LIU Dawei, HOU Senlin, ZHOU Yongwu, FEI Yiling, HE Jian. DNA barcoding of Artiodactyla in cases involved in wildlife for species identification[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(6): 27-32.

Figures/Tables 5

Table 1

Table 2

Table 3

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References 29

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物种 species	保护级别 protected category	样本数 number of samples	编号 No.	动物体部位 body part
斑羚Naemorhedus goral	II	3	NG1-3	角、肌肉
北山羊Capra ibex	I	5	CI1-5	角、肌肉
藏羚Pantholops hodgsonii	I	3	PH1-3	头骨
鹅喉羚Gazella subgutturosa	II	4	GS1-4	皮张
高鼻羚羊Saiga tatarica	I	6	ST1-6	角
河麂Hydropotes inermis	II	5	HI1-5	肌肉
黒麂 Muntiacus crinifrons	I	2	MC1-2	肌肉
鬣羚Capricornis sumatraensis	II	5	CS1-5	肌肉
羚牛 Budorcas taxicolor	I	3	BT1-3	头骨
马鹿Cervus elaphus	II	5	CE1-5	角
毛冠鹿Elaphodus cephalophus	三有	6	EC1-6	皮张
梅花鹿Cervus nippon	I	5	CN1-5	角
西伯利亚狍Capreolus pygargus	三有	6	CC1-6	肌肉
驼鹿Alces alces	II	3	AA1-3	角
小麂Muntiacus reevesi	三有	4	MR1-4	角、肌肉
马麝Moschus chrysogaster	I	2	MC'1-2	肌肉

物种 species	序列比对结果(相似度/%) sequences alignment result (similarities)
物种 species	相关物种COI基因片段 COI of concerned species	相关物种Cyt b基因片段 Cyt b of concerned species
斑羚	斑羚(99.75~100)	斑羚(98.74~100)
北山羊	北山羊(99.11~99.82)	北山羊(99.58~99.64)
藏羚	藏羚(99.47~99.82)	藏羚(99.47~99.82)
鹅喉羚	鹅喉羚(96.80~100)	鹅喉羚(99.29~99.82)
高鼻羚羊	高鼻羚羊(99.29~100)	高鼻羚羊(98.75~100)
河麂	河麂(98.04~99.64)	河麂(99.29~99.82)
黒麂	黒麂(99.47~100)	黒麂(98.93~99.11)
鬣羚	鬣羚(99.11~100)	鬣羚(98.75~100)
羚牛	羚牛(99.11~99.82)	羚牛(98.58~99.82)
马鹿	马鹿(97.51~99.64)	马鹿(98.76~99.64)
毛冠鹿	毛冠鹿(98.40~100)	毛冠鹿(98.40~100)
梅花鹿	梅花鹿(98.22~99.64)	梅花鹿(98.40~99.64)
西伯利亚狍	西伯利亚狍 (99.11~99.64)	西伯利亚狍、欧洲狍 (99.64~99.82)
驼鹿	驼鹿(98.75~100)	驼鹿(98.74~100)
小麂	小麂(96.98-99.82)	小麂(98.93~99.82)
马麝	马麝(98.93-99.29)	马麝(98.92~99.10)

DNA barcoding of Artiodactyla in cases involved in wildlife for species identification

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遗传距离 genetic distance	种内/% intra species		种间/% inter species
遗传距离 genetic distance	COI	Cyt b	COI	Cyt b
最小值maximum	0.0	0.0	1.8	4.0
最大值minimum	1.4	0.5	18.8	15.8
平均值mean	0.1	0.0	12.1	10.9
标准偏差S.D.	0.3	0.1	2.1	1.9

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