DNA条形码技术在青海海东地区小型兽类鉴定中的应用

doi:10.3724/SP.J.1003.2012.06188

生物多样性 ›› 2012, Vol. 20 ›› Issue (2): 193-198. DOI: 10.3724/SP.J.1003.2012.06188

所属专题：青藏高原生物多样性与生态安全

DNA条形码技术在青海海东地区小型兽类鉴定中的应用

马英¹, 李海龙¹, 鲁亮²^,^*(), 刘起勇²

1 青海省地方病预防控制所, 西宁 811602
2 中国疾病预防控制中心传染病预防控制所, 北京 102200

收稿日期:2011-10-20 接受日期:2012-03-10 出版日期:2012-03-20 发布日期:2012-04-09
通讯作者: 鲁亮
作者简介:* E-mail: luliang28@sohu.com
基金资助:
国家科技支撑计划资助项目(2008BAI56B02);国家科技重大项目(20082N0004-010);国家自然科学基金(31060279)

Application for identification of small mammals by DNA barcoding in Haidong area, Qinghai Province, China

Ying Ma¹, Hailong Li¹, Liang Lu²^,^*(), Qiyong Liu²

1 Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602
2 National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102200

Received:2011-10-20 Accepted:2012-03-10 Online:2012-03-20 Published:2012-04-09
Contact: Liang Lu

摘要/Abstract

摘要：

为弥补传统形态分类方法的不足, 探究应用DNA条形码技术进行分子生物学鉴定的可行性, 本研究用DNA条形码技术检测了青海省海东地区3目6科14属18种110只小型兽类的COI基因部分序列。分析所测COI基因序列可知: 种内遗传距离≤3%, 种间遗传距离5-10%, 属间遗传距离12-19%, 种间遗传距离显著大于种内遗传距离。NJ树显示同种个体聚为有很高支持度的单一分支。有6个个体(4只黄胸鼠、2只小家鼠)在现场鉴定中被误定为其他种类。研究结果表明使用条形码技术能纠正形态学鉴定中的错误, 也说明动物线粒体COI基因是一个有效的DNA条形码标准基因。

关键词: DNA条形码, 小兽, COI基因

Abstract

Because of the shortcomings of morphological classification, we studied the feasibility of DNA barcoding in the identification of small mammals. Small mammals belonging to 3 orders, 6 families and 14 genera were collected from the Haidong region in Qinghai Province. We analyzed COI gene sequences from 110 small mammal samples and the results showed that the average intraspecific genetic distance was small than 3%, average interspecific distance ranged from 5% to 10% and intergenus distance 12-19%. The average interspecific genetic distance was significantly greater than the intraspecific genetic distance. A Neighbor-Joining tree showed that samples of the same species formed monophyletic groups with high support value. The NJ tree implied that 6 individuals were misidentified in the field. Our results suggest that the DNA barcoding can increase identification accuracy relative to using morphology alone, and also that the mitochondrial COI gene is an efficacious candidate for DNA barcoding.

Key words: DNA barcoding, small mammals, COI gene

马英, 李海龙, 鲁亮, 刘起勇 (2012) DNA条形码技术在青海海东地区小型兽类鉴定中的应用. 生物多样性, 20, 193-198. DOI: 10.3724/SP.J.1003.2012.06188.

Ying Ma, Hailong Li, Liang Lu, Qiyong Liu (2012) Application for identification of small mammals by DNA barcoding in Haidong area, Qinghai Province, China. Biodiversity Science, 20, 193-198. DOI: 10.3724/SP.J.1003.2012.06188.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2012.06188

https://www.biodiversity-science.net/CN/Y2012/V20/I2/193

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参考文献 20

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物种 Species	样本来源 Source of samples	样本数量 Number of samples	序列数目 Number of sequence
啮齿目 Rodentia 仓鼠科 Circetidae 长尾仓鼠 Cricetulus longicaudatus	互助县、平安县、循化县、民和县 Huzhu, Ping’an, Xunhua and Minhe counties	14	14
高原松田鼠 Pitymys irene	互助县、平安县 Huzhu and Ping’an counties	6	6
柴达木根田鼠 Microtus limnophilus	互助县、平安县 Huzhu and Ping’an counties	9	9
洮州绒鼠 Caryomys eva	循化县 Xunhua County	5	5
甘肃鼢鼠 Myospalax cansus	互助县、民和县 Huzhu and Minhe counties	6	6
子午沙鼠 Meriones meridianus	民和县 Minhe County	5	5
鼠科 Muridae 黄胸鼠 Rattus tanezumi	循化县 Xunhua County	4	4
褐家鼠 R. norvegicus	互助县、循化县 Huzhu and Xunhua counties	4	4
社鼠 Niviventer confucianus	循化县 Xunhua County	4	4
小家鼠 Mus musculus	平安县、循化县 Ping’an and Xunhua counties	9	9
朝鲜姬鼠 Apodemus peninsulae	互助县、平安县、循化县、民和县 Huzhu, Ping’an, Xunhua and Minhe counties	13	13
黑线姬鼠 A. agrarius	民和县 Minhe County	2	2
松鼠科 Sciuridae
小飞鼠 Pteromys volans	互助县 Huzhu County	3	3
花鼠 Tamias sibiricus	循化县 Xunhua County	2	2
林跳鼠科 Zapodidae 中国蹶鼠 Sicista concolor	平安县 Ping’an County	2	2
兔形目 Lagomorpha
鼠兔科 Ochotonidae
甘肃鼠兔 Ochotona cansus	互助县、平安县、循化县 Huzhu, Ping’an and Xunhua counties	10	10
食虫目 Insectivora
鼩鼱科 Soricidae
山地背纹鼩鼱 Sorex bedfordiae	互助县、循化县 Huzhu and Xunhua counties	13	13
鼩鼱(待定种) Sorex sp.	循化县 Xunhua County	1	1

物种 Species	样本来源 Source of samples	样本数量 Number of samples	序列数目 Number of sequence
啮齿目 Rodentia 仓鼠科 Circetidae 长尾仓鼠 Cricetulus longicaudatus	互助县、平安县、循化县、民和县 Huzhu, Ping’an, Xunhua and Minhe counties	14	14
高原松田鼠 Pitymys irene	互助县、平安县 Huzhu and Ping’an counties	6	6
柴达木根田鼠 Microtus limnophilus	互助县、平安县 Huzhu and Ping’an counties	9	9
洮州绒鼠 Caryomys eva	循化县 Xunhua County	5	5
甘肃鼢鼠 Myospalax cansus	互助县、民和县 Huzhu and Minhe counties	6	6
子午沙鼠 Meriones meridianus	民和县 Minhe County	5	5
鼠科 Muridae 黄胸鼠 Rattus tanezumi	循化县 Xunhua County	4	4
褐家鼠 R. norvegicus	互助县、循化县 Huzhu and Xunhua counties	4	4
社鼠 Niviventer confucianus	循化县 Xunhua County	4	4
小家鼠 Mus musculus	平安县、循化县 Ping’an and Xunhua counties	9	9
朝鲜姬鼠 Apodemus peninsulae	互助县、平安县、循化县、民和县 Huzhu, Ping’an, Xunhua and Minhe counties	13	13
黑线姬鼠 A. agrarius	民和县 Minhe County	2	2
松鼠科 Sciuridae
小飞鼠 Pteromys volans	互助县 Huzhu County	3	3
花鼠 Tamias sibiricus	循化县 Xunhua County	2	2
林跳鼠科 Zapodidae 中国蹶鼠 Sicista concolor	平安县 Ping’an County	2	2
兔形目 Lagomorpha
鼠兔科 Ochotonidae
甘肃鼠兔 Ochotona cansus	互助县、平安县、循化县 Huzhu, Ping’an and Xunhua counties	10	10
食虫目 Insectivora
鼩鼱科 Soricidae
山地背纹鼩鼱 Sorex bedfordiae	互助县、循化县 Huzhu and Xunhua counties	13	13
鼩鼱(待定种) Sorex sp.	循化县 Xunhua County	1	1

引物名称 Primer name	引物序列(5′-3′) Primer sequence 5′-3′	引物间比例 Ratio
BatL5310	CCTACTCRGCCATTTTACCTATG	1
R6036R	ACTTCTGGGTGTCCAAAGAATCA	1
VF1LFt1
LepF1_t1	TGTAAAACGACGGCCAGTATTCAACCAATCATAAAGATATTGG	1
VF1_t1	TGTAAAACGACGGCCAGTTCTCAACCAACCACAAAGACATTGG	1
VF1d_t	TGTAAAACGACGGCCAGTTCTCAACCAACCACAARGAYATYGG	1
VF1i_t1	TGTAAAACGACGGCCAGTTCTCAACCAACCAIAAIGAIATIGG	3
VR1LRt1
LepRI_t1	CAGGAAACAGCTATGACTAAACTTCTGGATGTCCAAAAAATCA	1
VR1d_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGGCCRAARAAYCA	1
VR1_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGGCCAAAGAATCA	1
VR1i_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGICCIAAIAAICA	3

引物名称 Primer name	引物序列(5′-3′) Primer sequence 5′-3′	引物间比例 Ratio
BatL5310	CCTACTCRGCCATTTTACCTATG	1
R6036R	ACTTCTGGGTGTCCAAAGAATCA	1
VF1LFt1
LepF1_t1	TGTAAAACGACGGCCAGTATTCAACCAATCATAAAGATATTGG	1
VF1_t1	TGTAAAACGACGGCCAGTTCTCAACCAACCACAAAGACATTGG	1
VF1d_t	TGTAAAACGACGGCCAGTTCTCAACCAACCACAARGAYATYGG	1
VF1i_t1	TGTAAAACGACGGCCAGTTCTCAACCAACCAIAAIGAIATIGG	3
VR1LRt1
LepRI_t1	CAGGAAACAGCTATGACTAAACTTCTGGATGTCCAAAAAATCA	1
VR1d_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGGCCRAARAAYCA	1
VR1_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGGCCAAAGAATCA	1
VR1i_t1	CAGGAAACAGCTATGACTAGACTTCTGGGTGICCIAAIAAICA	3

DNA条形码技术在青海海东地区小型兽类鉴定中的应用

Application for identification of small mammals by DNA barcoding in Haidong area, Qinghai Province, China

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