基于形态学和DNA条形码的青藏高原东缘鼢鼠物种鉴定

doi:10.16829/j.slxb.150993

摘要/Abstract

摘要： 物种分类和鉴定是生物学研究的基石，综合分子生物学和经典分类学方法进行物种鉴定可提高结果可靠性。本研究采用传统形态学和线粒体细胞色素b基因（Cyt b）序列对青藏高原东缘青海省海东市平安区采集的74个鼢鼠样本进行物种鉴定。头骨形态鉴定结果显示，尾巴被稀疏短白毛的鼢鼠个体（Pac）为甘肃鼢鼠（Eospalax cansus），而尾巴被密毛的个体（Pab）为高原鼢鼠（Eospalax baileyi）。74个鼢鼠样本的Cyt b序列共定义了25个单倍型，结合GenBank中8种鼢鼠的Cyt b序列进行分析，在构建的系统发育树中，已知8种鼢鼠序列各自聚为1个单系群，平安区Pab的16个单倍型与已知高原鼢鼠聚为一支，Pac的8个单倍型与已知甘肃鼢鼠聚为一支。遗传距离结果显示，Pac与已知甘肃鼢鼠遗传距离小于与其他物种的遗传距离（P < 0.001），Pab与已知高原鼢鼠的遗传距离小于与其他物种的遗传距离（P < 0.001）。Cyt b基因分析和形态鉴定结果一致，Cyt b是鼢鼠鉴定的一个有效的DNA条形码基因。综合形态学和DNA条形码鉴定结果，平安区分布着2种鼢鼠：甘肃鼢鼠和高原鼢鼠。在地理分布上，甘肃鼢鼠主要分布在平安区北部邻近黄土高原海拔相对较低的区域，高原鼢鼠则分布在南部邻近青藏高原海拔相对较高的区域。本研究加深了对平安区鼢鼠物种组成和分布特征的了解，并为解决鼢鼠物种形态鉴定难的问题提供了DNA条形码解决方案。

关键词: DNA条形码技术, 鼢鼠, Cyt b, 形态, 系统发育, 遗传距离

Abstract: Species classification and identification constitute the cornerstone of biological research. Integrating DNA barcoding with classical taxonomic approaches can enhance the reliability of species identification. In this study, the traditional morphology and mitochondrial cytochrome b gene (Cyt b) sequence were employed to identify 74 zokor samples collected from Ping’an District, Haidong City, Qinghai Province, on the eastern of the Qinghai-Xizang Plateau. Based on skull morphology, individuals with sparsely coated short white hair on the tail (Pac) were identified as Eospalax cansus, whereas those with densely haired tail (Pab) were identified as Eospalax baileyi. A total of 25 Cyt b haplotypes were defined among the 74 specimens. Phylogenetic analysis based on Cyt b gene sequences of eight zokor species from GenBank showed that each species formed a well-supported monophyletic clade. The 16 haplotypes of Pab and the eight haplotypes of Pac from Ping’an District clustered within the clades of E. baileyii and E. cansu, respectively. The genetic distance between Pac and E. cansus was significantly smaller than that between Pac and other species (P < 0. 001), and similarly, the distance between Pab and E. baileyii was smaller than that between Pab and other species (P < 0. 001). The Cyt b identification results were consistent with those from skull morphology, confirming that the mitochondrial Cyt b is an effective DNA barcode candidate for Myospalacinae. Morphological and DNA barcoding results indicate that, two species, E. baileyii and E. cansus, are distributed in Ping’an District. Geographically, E. cansus occurs mainly at relatively low altitudes in the north, closer to the Loess Plateau, while E. baileyii is found at higher altitudes in the south, nearer the Qinghai-Xizang Plateau. This study improves our understanding of zokor species composition and distribution in Ping’an District, and provides a solution to the challenges of morphological identification in zokors through DNA barcoding.

Key words: DNA barcoding, zokor, Cyt b, Morphology, Phylogenetics, Genetic distance

中图分类号:

Q959

蔡振媛, 孔虹颖, 何振邦, 宋鹏飞, 郭凡, 李斌, 梁程博, 胡天石, 徐波, 林中原, 张同作. 基于形态学和DNA条形码的青藏高原东缘鼢鼠物种鉴定[J]. 兽类学报, 2026, 46(1): 116-128.

CAI Zhenyuan, KONG Hongying, HE Zhenbang, SONG Pengfei, GUO Fan, LI Bin, LIANG Chengbo, HU Tianshi, XU Bo, LIN Zhongyuan, ZHANG Tongzuo. Species identification of zokors in the east of the Qinghai-Xizang Plateau based on both morphology and DNA barcoding[J]. ACTA THERIOLOGICA SINICA, 2026, 46(1): 116-128.

参考文献

Bandelt H J,Forster P,Rohl A,1999. Median-joining networks for inferring intraspecific phylogenies[J]. Molecular Biology and Evolution, 16:37-48. DOI:10. 1093/oxfordjournals. molbev.a026036.
Bickford D,Lohman D J,Sodhi N S,Ng P K,Meier R,Winker K,Ingram K K,Das I,2007. Cryptic species as a window on diversity and conservation[J]. Trends in Ecology & Evolution,22(3):148-155. DOI:10. 1016/j. tree. 2006. 11. 004.
Candek K, Kuntner M, 2015. DNA barcoding gap:Reliable species identification over morphological and geographical scales[J]. Molecular Ecology Resources, 15:268-277. DOI:10. 1111/1755-0998. 12304.
Fan Naichang,Shi Yinzhu,1982. A revision of the zokors of subgenus Eospalax[J]. Acta Theriologica Sinica, 2 (2):183-199.(in Chinese with English abstract)
Felsenstein J,1985. Confidence limits on phylogenies:an approach using the bootstrap[J]. Evolution, 39 (4):783-791. DOI:10. 2307/2408678.
Gao Lianming,2015. Applications of DNA barcoding in biodiversity inventory and assessment[J]. Biodiversity Science, 23 (3):286-287. DOI:10. 17520/biods. 2015070. (in Chinese with English abstract)
Ginestet C, 2011. ggplot2:Elegant graphics for data analysis[J]. Journal of the Royal Statistical Society Series A:Statistics in Society, 174 (1):245-246. DOI:10. 1111/j. 1467-985X. 2010. 00676_9. x.
Guo Limin,Xin Jinxiao,Shi Zhanwen,Zhang Fenchun,Jiang Haiwei,Bai Zhonghui,2019. Evaluation on effect of DNA barcoding on identification of rodent species in Spermophilus alaschanicus plague foci of Pingchuan district[J]. Bulletin of Disease Control & Prevention (China),34 (6):1-3,24. DOI:10. 13215/j.cnki. jbyfkztb. 1911001. (in Chinese with English abstract)
He Kai, Wang Wenzhi, Li Quan, Luo Peipeng, Sun Yuehua, Jiang Xuelong, 2013. DNA barcoding in surveys of small mammal community:a case study in Lianhuashan,Gansu Province,China[J]. Biodiversity Science,21(2):197-205. DOI:10. 3724/SP. J. 1003. 2013. 09160. (in Chinese with English abstract)
Hebert P D,Cywinska A,Ball S L,Dewaard J R,2003. Biological identifications through DNA barcodes[J]. Proceedings of the Royal Society of London Series B:Biological Sciences, 270(1512):313-321. DOI:10. 1098/rspb. 2002. 2218.
Jiang F,Jin Q,Liang L,Zhang A B,Li Z H,2014. Existence of species complex largely reduced barcoding success for invasive species of Tephritidae:a case study in Bactrocera spp.[J]. Molecular Ecology Resources, 14:1114-1128. DOI:10. 1111/1755-0998. 12259.
Kimura M,1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences[J]. Journal of Molecular Evolution,16 (2):111-120.DOI:10. 1007/BF01731581.
Kress W J, GarcíA-Robledo C, Uriarte M, Erickson D L, 2015. DNA barcodes for ecology, evolution, and conservation[J]. Trends in Ecology & Evolution,30 (1):25-35. DOI:10. 1016/j. tree. 2014. 10. 008.
Leasi F,Sevigny J L,Laflamme E M,Artois T,Curini-Galletti M,De Jesus Navarrete A, Di Domenico M, Goetz F, Hall J A, Hochberg R, JöRger K M, 2018. Biodiversity estimates and ecological interpretations of meiofaunal communities are biased by the taxonomic approach[J]. Communications Biology, 1(1):112. DOI:10. 1038/s42003-018-0119-2.
Li Baoguo,Chen Fuguan,1992. A taxonomic revision of the subgenus Eospalax genus Myospalax[J]. Journal of Northwest University (Natural Science Edition),22 (3):339-344. (in Chinese with English abstract)
Li Xiaochen,Wang Tingzheng,1996. Taxonomy and phylogeny of subgenus Eospalax[J]. Journal of Shaanxi Normal University(Natural Science Edition), 24 (3):79-82. (in Chinese with English abstract)
Lin Y H, Waddell P J, Penny D, 2002. Pika and vole mitochondrial genomes increase support for both rodent monophyly and glires[J]. Gene, 294 (1-2):119-129. DOI:10. 1016/S0378-1119 (02) 00695-9.
Liu D X, Yan J Y, Wang H J, Jiang F, Song P F, Cai Z Y,Zhang T Z,2021. Microbial biogeography along the gastrointestinal tract segments of sympatric subterranean rodents (Eospalax baileyi and Eospalax cansus)[J]. Animals, 11 (11):3297.DOI:10. 3390/ani11113297.
Liu J L,Dujsebayeva T N,Chirikova M A,Gong X,Li D J,Guo X G, 2021. Does the Dzungarian racerunner (Eremias dzungarica Orlova,Poyarkov,Chirikova,Nazarov,Munkhbaatar,Munkhbayar & Terbish,2017) occur in China? Species delimitation and identification with DNA barcoding and morphometric analyses[J]. Zoological Research, 42 (3):287. DOI:10. 24272/j. issn. 2095-8137. 2020. 318.
Liu Li, Zhou Yanshan, Chu Bin, Wang Guizhen, Hua Limin, 2018. Classification of two zokor species based on mitochondrial gene, morphological and habitat indices[J]. Acta Theriologica Sinica, 38 (4):402-410. DOI:10. 16829/j. slxb. 150188.(in Chinese with English abstract)
Lou Qian,Xin Tianyi,Song Jingyuan,2020. Application of DNA barcoding technology in the whole industrial chain of traditional Chinese medicine[J]. Acta Pharmaceutica Sinica, 55 (8):1784-1791. DOI:0. 16438/j. 0513-4870. 2020-0290. (in Chinese with English abstract)
Norris R W,Zhou K Y,Zhou C Q,Yang G,Kipatrick C W,Honeycutt R L, 2004. The phylogenetic position of the zokors(Myospalacinae) and comments on the families of muroids(Rodentia)[J]. Molecular Phylogenetics and Evolution, 31(3):972-978. DOI:10. 1016/j. ympev. 2003. 10. 020.
Rozas J, Ferrer-Mata A, SáNchez-Delbarrio J C, Guirao-Rico S,Librado P, Ramos-Onsins S E, SáNchez-Gracia A, 2017.DnaSP 6:DNA sequence polymorphism analysis of large data sets[J]. Molecular Biology and Evolution, 34 (12):3299-3302. DOI:10. 1093/molbev/msx248.
Tamura K,Stecher G,Kumar S,2021. MEGA11:molecular evolutionary genetics analysis version 1l[J]. Molecular Biology and Evolution, 38 (7):3022-3027. DOI:10. 1093/molbev/msab120.
Thompson J D,Gibson T J,Plewniak F,Jeanmougin F,Higgins D G,1997. The CLUSTAL X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 25 (24):4876-4882. DOI:10. 1093/NAR/25. 24. 4876.
Wang Chan, Pan Yongtai, Wang Jing, Liu Fayang, Yao Baohui,Su Junhu, 2020. Selection of plateau zokor (Eospalax baileyi) DNA barcodes[J]. Pratacultural Science, 37 (12):2574-2583. DOI:10. 11829/j. issn. 1001-0629. 2020-0054. (in Chinese with English abstract)
Wang Jialian, Yang Guang, Liu Hai, Zhou Kaiya, Wei Fuwen, 2003. Application of mitochondrial DNA sequences in the species identification of common dolphins (Genus Delphinus) in Chinese waters[J]. Acta Theriologica Sinica, 23 (2):120-126. DOI:10. 16829/j. slxb. 2003. 02. 006. (in Chinese with English abstract)
Xing Chao,Lin Yi,Zhou Zhiqiang,Zhao Lianjun,Jiang Shiwei,Lin Zhenzhen,Xu Jiliang,Zhan Xiangjiang,2023. The establishment of terrestrial vertebrate genetic resource bank and species identification base on DNA barcoding in Wanglang National Nature Reserve[J]. Biodiversity Science,31 (7):87-99. DOI:10. 17520/biods. 2022661. (in Chinese with English abstract)
Yang Qianqian,Liu Suwen,Yu Xiaoping,2018. Research progress on DNA barcoding analysis methods[J]. Chinese Journal of Applied Ecology, 29 (3):1006-1014. DOI:10. 13287/j. 1001-9332. 201803. 032. (in Chinese with English abstract)
Zhang Caiyun, Huang Shanshan, Yan Haifei, 2017. Applications of DNA barcoding in Chinese materia medica identification[J]. Chinese Traditional and Herbal Drugs, 48 (11):2306-2312.DOI:10. 7501/j. issn. 0253-2670. 2017. 11. 026. (in Chinese with English abstract)
Zhao Fang, Deng Xiaogong, Zhang Tongzuo, Su Jianping, Lin Gonghua, 2015. Molecular authentication of Sailonggu and its resource distribution in Qinghai-Xizang Plateau[J]. China Journal of Chinese Materia Medica, 40 (3):399-403. DOI:10. 4268/cjcmm20150306. (in Chinese with English abstract)
Zhao Xianxian, Li Bang, Lin Gonghua, Ma Wanjun, Ju Hailan,Su Jianping,Zhang Tongzuo,2017. Molecular identification of voles in the east of the Qinghai-Xizang Plateau[J]. Acta Theriologica Sinica, 37 (1):44-52. DOI:10. 16829/j.slxb. 201701002. (in Chinese with English abstract)
Zhou C Q, Zhou K Y, Zhang S L, 2004. Molecular authentication of the animal crude drug Sailonggu (bone of Myospalax baileyi)[J]. Biological and Pharmaceutical Bulletin,27 (11):1850. DOI:10. 1248/bpb. 27. 1850.
Zhu Lin, Li Ting, Chen Bingyao, Yang Guang, 2020. The morphological and mtDNA molecular identification of Fin whale and Omura's whale[J]. Acta Theriologica Sinica,40 (1):37-46.DOI:10. 16829/j. slxb. 150349. (in Chinese with English abstract)
王加连,杨光,刘海,周开亚,魏辅文,2003. 线粒体 DNA 序列分析在中国水域真海豚物种鉴定中的初步应用[J]. 兽类学报, 23 (2):120-126. DOI:10. 16829/j. slxb. 2003. 02. 006.
王缠,潘永泰,王静,刘发央,姚宝辉,苏军虎,2020. 高原鼢鼠DNA 条形码筛选[J]. 草业科学,37 (12):2574-2583. DOI:10. 11829/j. issn. 1001-0629. 2020-0054.
中国科学院西北高原生物研究所,1989. 青海经济动物志[M]. 西宁:青海人民出版社.
邢超,林依,周智强,赵联军,蒋仕伟,林蓁蓁,徐基良,詹祥江,2023. 基于 DNA 条形码技术构建王朗国家级自然保护区陆生脊椎动物遗传资源数据库及物种鉴定[J]. 生物多样性,31(7):87-99. DOI:10. 17520/biods. 2022661.
朱林,李婷,陈炳耀,杨光,2020. 利用形态学及 mtDNA 基因分子鉴定长须鲸和大村鲸[J]. 兽类学报,40 (1):37-46. DOI:10. 16829/j. slxb. 150349.
刘丽,周延山,楚彬,王贵珍,花立民,2018. 基于线粒体基因、形态学和栖息地指标的两种鼢鼠分类研究[J]. 兽类学报,38(4):402-410. DOI:10. 16829/j. slxb. 150188.
李保国,陈服官,1992. 鼢鼠属凸颅亚属的分类整理[J]. 西北大学学报 (自然科学版),22 (3):339-344.
李晓晨,王廷正,1996. 论鼢鼠属 Eospalax 亚属的分类及系统演化[J]. 陕西师范大学学报 (自然科学版),24 (3):79-82.
杨倩倩, 刘苏汶, 俞晓平, 2018. DNA 条形码分析方法研究进展[J]. 应用生态学报, 29 (3):1006-1014. DOI:10. 13287/j. 1001-9332. 201803. 032.
何锴,王文智,李权,罗培鹏,孙悦华,蒋学龙,2013. DNA 条形码技术在小型兽类鉴定中的探索:以甘肃莲花山为例[J]. 生物多样性, 21 (2):197-205. DOI:10. 3724/SP. J. 1003. 2013. 09160.
张彩云,黄珊珊,颜海飞,2017. DNA 条形码技术在中药鉴定中的应用进展[J]. 中草药,48 (11):2306-2312. DOI:10. 7501/j. issn. 0253-2670. 2017. 11. 026.
赵芳,邓小弓,张同作,苏建平,林恭华,2015. 青藏高原地区塞隆骨资源的分子鉴定和地理分布[J]. 中国中药杂志,40 (3):399-403. DOI:10.4268/cjcmm20150306.
赵贤贤,李邦,林恭华,马万军,巨海兰,苏建平,张同作, 2017. 青藏高原东部地区田鼠物种的分子鉴定[J]. 兽类学报, 37 (1):44-52. DOI:10. 16829/j. slxb. 201701002.
娄千,辛天怡,宋经元,2020. DNA 条形码技术在中药全产业链的应用进展[J]. 药学学报, 55 (8):1784-1791. DOI:0. 16438/j. 0513-4870. 2020-0290.
高连明, 2015. DNA 条形码在生物多样性编目与评价中的应用[J]. 生物多样性, 23 (3):286-287. DOI:10. 17520/biods. 2015070.
郭丽民,席进孝,师占文,张奋春,姜海威,白仲慧,2019.DNA 条形码对平川区阿拉善黄鼠疫源地鼠种鉴定的效果评价[J]. 疾病预防控制通报,34 (6):1-3,24. DOI:10. 13215/j. cnki. jbyfkztb. 1911001.
樊乃昌,施银柱,1982. 中国鼢鼠 (EOSPALAX) 亚属分类研究[J]. 兽类学报,2 (2):183-199.

[1]	刘铸, 张璐, 郭秋颖, 张智慧, 蒋纹静, 田新民, 金志民, 张隽晟. 分布于贵州的一种小麝鼩分类地位的厘定[J]. 兽类学报, 2025, 45(4): 537-547.
[2]	蒲小燕, 戴重阳, 王雅轩, 魏登邦. 低氧环境下高原鼢鼠及SD大鼠肺和骨骼肌中SP‑A的表达模式[J]. 兽类学报, 2025, 45(3): 283-294.
[3]	阿布都克尤木·卡德尔, 孙婷, 王大伟, 刘晓辉, 汪秀会, 宋英. 抗凝血剂对褐家鼠种群遗传多样性及抗药性的影响[J]. 兽类学报, 2025, 45(1): 78-85.
[4]	陈顺德, 杨思雨, 青娇, 范荣辉, 唐刻意, 廖锐, 郭克疾, 张梦斐, 吴南飞, 刘少英. 中国西藏发现麝鼩属(劳亚食虫目：鼩鼱科)两新物种[J]. 兽类学报, 2024, 44(5): 529-550.
[5]	刘莹洵, 彭步青, 王旭明, 廖锐, 潘璇, 刘少英. 中国兽类一新纪录：尼泊尔长尾鼩鼱[J]. 兽类学报, 2024, 44(5): 551-557.
[6]	李玉鹏, 戴宇飞, 吴福星, 祝茜. 福建长乐一头搁浅柏氏中喙鲸的形态学记录[J]. 兽类学报, 2024, 44(4): 506-514.
[7]	高艺, 吴禹岐, 韩美凤, 郭秋颖, 蔡赫, 张春凤, 金志民, 陈欢, 张隽晟, 刘铸. 长尾蹶鼠形态学和分子系统学初步分析[J]. 兽类学报, 2024, 44(4): 515-522.
[8]	祝常悦, 钟旭凯, 王昱心, 李文永, 齐旭明, 范朋飞. 海南长臂猿的毛色变化[J]. 兽类学报, 2024, 44(1): 1-13.
[9]	张佳钰, 安志芳, 王志洁, 陈晓琦, 魏登邦. 高海拔环境抑制高原鼢鼠和高原鼠兔的胆汁酸代谢[J]. 兽类学报, 2023, 43(5): 553-567.
[10]	杨思雨, 谢菲, 唐刻意, 付长坤, 刘洋, 刘少英, 陈顺德. 福建、浙江和湖南三省兽类新纪录——安徽麝鼩[J]. 兽类学报, 2023, 43(5): 620-625.
[11]	刘铸, 钱洪安, 王再山, 韩美凤, 蔡赫, 张春凤, 时艳菁, 金志民, 陈欢, 张隽晟. 基于Cyt b基因和形态学探讨大麝鼩的亚种分化[J]. 兽类学报, 2023, 43(4): 422-429.
[12]	刘强, 王海军, 赵越, 陈世玉, 姜广顺. 中国东北豹头骨三维重建及测量参数[J]. 兽类学报, 2023, 43(3): 315-321.
[13]	覃雯, 杨传华, 蔡振媛. 高原鼢鼠和甘肃鼢鼠胆汁无机元素含量分析[J]. 兽类学报, 2023, 43(2): 229-234.
[14]	安康, 包明芳, 姚宝辉, 康宇坤, 谭宇尘, 王艳莉, 苏军虎. MTNR1a和MTNR1b基因在繁殖期与非繁殖期雄性高原鼢鼠HPG轴上的表达[J]. 兽类学报, 2022, 42(4): 410-419.
[15]	谢菲, 万韬, 唐刻意, 王旭明, 陈顺德, 刘少英. 中国拟家鼠分类与分布厘订[J]. 兽类学报, 2022, 42(3): 270-285.