DNA条形码技术鉴定浙江省常见鼠形动物研究
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摘要
目的利用DNA条形码技术鉴定浙江省常见鼠形动物。方法在浙江省嘉善县、龙游县、云和县和宁海县捕获野外鼠形动物,取鼠耳标本提取DNA,采用通用引物PCR扩增线粒体细胞色素C氧化酶亚基I (COI)基因测序。将测序获得序列与Gen Bank中的序列进行Blast序列同源性比对,采用邻接法构建分子进化树鉴定鼠形动物,并与传统形态学方法鉴定结果进行比较。结果共扩增出22份COI标本,其中18份标本构建的进化树与传统形态学鉴定结果一致;3份标本的鉴定结果存在差异,臭鼩鼱(Suncus murinus)重新鉴定为大麝鼩(Crocidura lasiura),幼年黄毛鼠(Rattus losea)、幼年黄胸鼠(Rattus tanezumi)重新鉴定为幼年褐家鼠(Rattus rattus);1份形态学鉴定为幼年东方田鼠(Microtus fortis)的标本(编号:NH-1)在Gen Bank中同源性较低,有待进一步鉴定。结论 DNA条形码技术能有效更正传统形态学鉴定可能造成的错误,两种方法联合可提高鼠型动物鉴定的准确性。
Objective To apply DNA barcoding to identifying the rodents in Zhejiang Province. Methods Rodents were captured from Jiashan,Longyou,Yunhe and Ninghai counties in Zhejiang Province. The DNA was extracted from ears of rodent samples, and was amplified and sequenced with mitochondrial cytochrome C oxidase subunit I(COI) genes. The obtained sequences were compared with the related sequences in Gen Bank,and neighbour-joining evolutionary tree was constructed. Then the results by DNA barcoding and by morphological identification were compared. Results A total of 22 COI gene samples were amplified. The evolutionary tree constructed by 18 samples was consistent with the morphological identification results and 4 samples were different: Suncus murinus should be Crocidura lasiura, infant rats of Rattus losea and Rattus tanezumi was re-identified as Rattus rattus,infant rats of Microtus fortis(sample number:NH-1) needs further identification. Conclusion DNA barcoding can effectively correct the errors of morphological identification,thus combining the two methods could improve the accuracy of rodent identification.
Objective To apply DNA barcoding to identifying the rodents in Zhejiang Province. Methods Rodents were captured from Jiashan,Longyou,Yunhe and Ninghai counties in Zhejiang Province. The DNA was extracted from ears of rodent samples, and was amplified and sequenced with mitochondrial cytochrome C oxidase subunit I(COI) genes. The obtained sequences were compared with the related sequences in Gen Bank,and neighbour-joining evolutionary tree was constructed. Then the results by DNA barcoding and by morphological identification were compared. Results A total of 22 COI gene samples were amplified. The evolutionary tree constructed by 18 samples was consistent with the morphological identification results and 4 samples were different: Suncus murinus should be Crocidura lasiura, infant rats of Rattus losea and Rattus tanezumi was re-identified as Rattus rattus,infant rats of Microtus fortis(sample number:NH-1) needs further identification. Conclusion DNA barcoding can effectively correct the errors of morphological identification,thus combining the two methods could improve the accuracy of rodent identification.
引文
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[15]陈宝宝,孙养信,安翠红,等.用DNA条形码技术鉴定阿拉善黄鼠和达乌尔黄鼠[J].中国媒介生物学及控制杂志,2014,25(5):432-434.
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[17]刘润吉,张荣波,郭天宇,等. DNA条形码技术在贵州茂兰鼠类鉴定中的应用[J].中华卫生杀虫药械,2014(1):59-62.
[18]刘勇,宋毓,李晓宇.基于线粒体COI基因的DNA条形码技术在昆虫分子鉴定中的应用[J].植物检疫,2010,24(2):46-50.
[19] SHEN Y Y,XIAO C,MURPHY R W. Assessing DNA barcoding as a tool for species identification and data quality control[J].PLo S One,2013,8(2):e57125.
[2]汤永康,傅桂明,黄良,等.浙江省2003年鼠类种群分布监测[J].预防医学,2004,16(12):50.
[3]闫东,鲁亮,金圣浩,等.DNA条形码技术在河北省鼠疫疫源地鼠种鉴定中的应用[J].中国媒介生物学及控制杂志,2014,25(1):21-23.
[4]王安东,尹小平,田延河,等. DNA条形码技术在中国-哈萨克斯坦边境阿拉山口地区鼠类鉴定中的应用[J].中国媒介生物学及控制杂志,2016,27(5):459-462.
[5]刘蓉蓉,葛德燕,鲁亮,等.中国姬鼠属种类的DNA条形码鉴定及其分布[J].中国媒介生物学及控制杂志,2017,28(2):97-103.
[6] HEBERT P D N, CYWINSKA A, BALL S L, et al. Biological identifications through DNA barcodes[J]. Proceedings Biological Sciences,2003,270(1512):313-321.
[7] HEBERT P D N,RATNASINGHAM S,WAARD J R D. Barcoding animal life:cytochrome c oxidase subunit I divergences among closely related species[J]. Proc Biol Sci,2003,270(S1):96-99.
[8] HEBERT P D N,PENTON E H,BURNS J M,et al. Ten species in one:DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator[J]. Proceedings of the National Academy of Sciences of the United States of America,2004,101(41):14812-14817.
[9]肖永刚,陈祥盛.基于线粒体COI基因的17种菱蜡蝉亚科昆虫DNA条形码研究(半翅目:蜡蝉总科:菱蜡蝉科)[J].山地农业生物学报,2014,33(2):44-50.
[10]何锴,王文智,李权,等. 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,2013,21(2):197-205.
[11]马英,李海龙,鲁亮,等. DNA条形码技术在青海海东地区小型兽类鉴定中的应用[J].生物多样性,2012,20(2):193-198.
[12]岳巧云,邱德义,胡佳,等. DNA条形码-医学媒介生物快速准确鉴定的利器[J].检验检疫学刊,2013(5):60-63.
[13]常子丽,刘芳,王建军,等. DNA条形码鉴别内蒙古地区啮齿动物[J].生物技术通报,2013(8):94-98.
[14]马英,李海龙,李超,等.基于DNA条形码技术鉴定青海柴达木盆地小型兽类[J].中国人兽共患病学报,2013,29(12):1186-1190.
[15]陈宝宝,孙养信,安翠红,等.用DNA条形码技术鉴定阿拉善黄鼠和达乌尔黄鼠[J].中国媒介生物学及控制杂志,2014,25(5):432-434.
[16]陈炼,吴琳,王启菲,等. DNA条形码及其在生物多样性研究中的应用[J].四川动物,2016,35(6):942-949.
[17]刘润吉,张荣波,郭天宇,等. DNA条形码技术在贵州茂兰鼠类鉴定中的应用[J].中华卫生杀虫药械,2014(1):59-62.
[18]刘勇,宋毓,李晓宇.基于线粒体COI基因的DNA条形码技术在昆虫分子鉴定中的应用[J].植物检疫,2010,24(2):46-50.
[19] SHEN Y Y,XIAO C,MURPHY R W. Assessing DNA barcoding as a tool for species identification and data quality control[J].PLo S One,2013,8(2):e57125.