上海口岸截获的粉蝇属DNA条形码鉴定及系统发育研究
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摘要
使用传统形态学分类方法进行干制标本的物种鉴定通常比较困难。本文按照《国境口岸医学媒介昆虫DNA条形码鉴定操作规程》对上海口岸截获的粉蝇属干制标本进行DNA条形码研究。经BOLD System v4比对,结果显示所得序列分别属于5个种:粗野粉蝇Pollenia rudis、伪粗野粉蝇P.pediculata、游荡粉蝇P.vagabunda、匈牙利粉蝇P.hungarica和Pollenia sp.。引入1条COⅠ序列作为外群对全部序列构建系统发育树(NJ树),结果显示不同物种各自聚为一支。因此,COⅠ条形码技术可以弥补传统形态学分类方法的不足,提高蝇类干制标本鉴定的成功率。
Species identification with dry specimen is usually difficult by traditional taxonomy method. According to the operational regulations for DNA barcode identification of medical insects at frontier ports, DNA barcodes of 31 dry specimens of Pollenia species captured at Shanghai port were generated. After comparing with the Barcode of Life Data(BOLD) system v4, all the 31 DNA barcodes are assigned into 5 species: Pollenia rudis(Fabricius, 1794), Pollenia pediculata(Macquart, 1834), Pollenia vagabunda(Meigen, 1826), Pollenia hungarica(Rognes, 1987) and Pollenia sp.. A neighbor-joining tree was created based on the 31 barcodes of Pollenia and an inductive COⅠ sequence as an outgroup, it is revealed that these species can be differentiated by DNA barcodes. Therefore, COⅠ barcoding can overcome the shortages of traditional taxonomy, and improve the success rate in identification of flies with dry specimen.
Species identification with dry specimen is usually difficult by traditional taxonomy method. According to the operational regulations for DNA barcode identification of medical insects at frontier ports, DNA barcodes of 31 dry specimens of Pollenia species captured at Shanghai port were generated. After comparing with the Barcode of Life Data(BOLD) system v4, all the 31 DNA barcodes are assigned into 5 species: Pollenia rudis(Fabricius, 1794), Pollenia pediculata(Macquart, 1834), Pollenia vagabunda(Meigen, 1826), Pollenia hungarica(Rognes, 1987) and Pollenia sp.. A neighbor-joining tree was created based on the 31 barcodes of Pollenia and an inductive COⅠ sequence as an outgroup, it is revealed that these species can be differentiated by DNA barcodes. Therefore, COⅠ barcoding can overcome the shortages of traditional taxonomy, and improve the success rate in identification of flies with dry specimen.
引文
Chen QG,Ding LM,Wang BR,Zhou QM,Zhong T.Investigation report on vector insects in inbound containers at Shanghai Port.Chinese Journal of Frontier Health and Quarantine,1996,19(5):276-277.[陈清国,丁鲁民,王宝荣,周启明,钟韬.上海口岸入境集装箱媒介昆虫调查报告.中国国境卫生检疫杂志,1996,19(5):276-277.]
Fan ZD.A key of common flies in China.2nd ed.Beijing:Science Press,1992,524.[范滋德主编.中国常见蝇类检索表.第2版.北京:科学出版社,1992,524.]
Faulde M,Sobe D,Burghardt H,Wermter R.Hospital infestation by the cluster fly,Pollenia rudis sensu stricto Fabricius 1794 (Diptera:Calliphoridae),and its possible role in transmission of bacterial pathogens in Germany.International Journal of Hygiene and Environmental Health,2001,203(3):201-204.
Folmer O,Black M,Hoeh W,Lutz R,Vrijenhoek R.DNA primers for amplification of mitochondrial cytochrome C oxidase subunit I from diverse metazoan invertebrates.Mol Mar Biol Biotechnol.,1994,3(5):294-299.
Gao B,Wang YP,Xu Q,Zheng FF.COⅠ Sequences analysis for three kinds of mosquitoes at Fujian Port.Chinese Frontier Health Quarantine,2010,33(6):398-410.[高博,王宇平,许卿,郑芳芳.福建口岸三蚊种COⅠ基因序列分析.中国国境卫生检疫杂志,2010,33(6):398-410.]
Jewiss-Gaines A,Marshall SA,Whitworth TL.Cluster flies (Calliphoridae:Polleniinae:Pollenia) of North.Canadian Journal of Arthropod Identification,2012(19):1-4.
Li HF.Study on risk analysis of health and quarantine and countermeasure of entry containers.Port Health Control,2005,10(5):11-13.[李海峰.进境集装箱卫生检疫风险分析与对策研究.口岸卫生控制,2005,10(5):11-13.]
Hebert PDN,Cywinska A,Ball SL,deWaard JR.Biological identifications through DNA barcodes.Royal Society,2003,270(1 512):313-321.
Hebert PDN,Ratnasingham S,deWaard JR.Barcoding animal life:cytochrome C oxidase subunit 1 divergences among closely related species.Proceedings of the Royal Society B:Biological Sciences,2003,270(Suppl 1):96-99.
Pereira HM et al.Scenarios for global biodiversity in the 21st century.Science,2010,330(6010):1 496-1 450.
Rognes K.Blowflies (Diptera:Calliphoridae) of Fennoscandia and Denmark.Fauna Entomol.Scand,1991,24:1-272.
Rognes K.Revision of the cluster-flies of the Pollenia venturii species-group,with a cladistic analysis of Palaearctic species of Pollenia Robineau-Desvoidy (Diptera:Calliphoridae).Insect Systematics & Evolution,1992,23(3):233-248.
Rognes K.The taxonomy of the Pollenia rudis species-group in the Holarctic Region (Diptera:Calliphoridae).Systematic Entomology,1987,12(4):475-502.
Shi YX,Xiang DP,Li ZH,Hong Y,Li XB,Xing LQ,Guo BX,Huang JC,Zheng K.COⅠ sequences analysis and molecular identification for five species of mosquitoes from Guangdong Port.Chinese Frontier Health Quarantine,2008,31(2):103-107.[师永霞,相大鹏,李祖海,洪烨,李小波,幸芦琴,郭波旋,黄吉城,郑夔.广东国境口岸不同蚊种COⅠ序列分析和分子鉴定方法.中国国境卫生检疫杂志,2008,31(2):103-107.]
Wei XY,Qiu DY,Yue QY,Liu DX.DNA barcoding and morphology identification of Pollenia rudis-a non-recorded fly species in China.Chinese Journal of Vector Biology and Control,2016,27(4):326-329.[魏晓雅,邱德义,岳巧云,刘德星.粗野粉蝇的DNA条形码和形态鉴定.中国媒介生物学及控制杂志,2016,27(4):326-329.]
Wu W,Wang FR,Xia DF.Pollenia pediculata Macquart intercepted from exotic empty containers (Diptera:Calliphoridae).Acta Parasitology et Medica Entomologica Sinica,2009,16(2):126-128.[吴薇,王芙蓉,夏德峰.入境空箱中截获的伪粗野粉蝇(双翅目:丽蝇科).寄生虫与医学昆虫学报,2009,16(2):126-128.]
Yang QG,Cai J,Song GR,Deng YH,Zhou L,He J,Hu SS,Nei GA,Sun LX.Morphology and DNA barcoding identification of Pollenia Labialis intercepted at China Port.Acta Parasitology et Medica Entomologica Sinica,2017,(3):169-173.[杨庆贵,蔡佳,宋桂荣,邓耀华,周磊,何江,胡双双,聂国嫒,孙立新.中国口岸首次截获长阳粉蝇的形态及DNA条形码鉴定.寄生虫与医学昆虫学报,2017,(3):169-173.]
Yue QY,Qiu DY,Huang YW,Hou J,Liu GX,Hu LF.Application of DNA barcoding techniques to the species identification of unknown Diptera pupa.Chinese Frontier Health Quarantine,2011,34(5):343-347.[岳巧云,邱德义,黄艺文,侯捷,刘国雄,胡龙飞.应用DNA条形码技术鉴定未知双翅目蛹.中国国境卫生检疫杂志.2011,34(5):343-347.]
Fan ZD.A key of common flies in China.2nd ed.Beijing:Science Press,1992,524.[范滋德主编.中国常见蝇类检索表.第2版.北京:科学出版社,1992,524.]
Faulde M,Sobe D,Burghardt H,Wermter R.Hospital infestation by the cluster fly,Pollenia rudis sensu stricto Fabricius 1794 (Diptera:Calliphoridae),and its possible role in transmission of bacterial pathogens in Germany.International Journal of Hygiene and Environmental Health,2001,203(3):201-204.
Folmer O,Black M,Hoeh W,Lutz R,Vrijenhoek R.DNA primers for amplification of mitochondrial cytochrome C oxidase subunit I from diverse metazoan invertebrates.Mol Mar Biol Biotechnol.,1994,3(5):294-299.
Gao B,Wang YP,Xu Q,Zheng FF.COⅠ Sequences analysis for three kinds of mosquitoes at Fujian Port.Chinese Frontier Health Quarantine,2010,33(6):398-410.[高博,王宇平,许卿,郑芳芳.福建口岸三蚊种COⅠ基因序列分析.中国国境卫生检疫杂志,2010,33(6):398-410.]
Jewiss-Gaines A,Marshall SA,Whitworth TL.Cluster flies (Calliphoridae:Polleniinae:Pollenia) of North.Canadian Journal of Arthropod Identification,2012(19):1-4.
Li HF.Study on risk analysis of health and quarantine and countermeasure of entry containers.Port Health Control,2005,10(5):11-13.[李海峰.进境集装箱卫生检疫风险分析与对策研究.口岸卫生控制,2005,10(5):11-13.]
Hebert PDN,Cywinska A,Ball SL,deWaard JR.Biological identifications through DNA barcodes.Royal Society,2003,270(1 512):313-321.
Hebert PDN,Ratnasingham S,deWaard JR.Barcoding animal life:cytochrome C oxidase subunit 1 divergences among closely related species.Proceedings of the Royal Society B:Biological Sciences,2003,270(Suppl 1):96-99.
Pereira HM et al.Scenarios for global biodiversity in the 21st century.Science,2010,330(6010):1 496-1 450.
Rognes K.Blowflies (Diptera:Calliphoridae) of Fennoscandia and Denmark.Fauna Entomol.Scand,1991,24:1-272.
Rognes K.Revision of the cluster-flies of the Pollenia venturii species-group,with a cladistic analysis of Palaearctic species of Pollenia Robineau-Desvoidy (Diptera:Calliphoridae).Insect Systematics & Evolution,1992,23(3):233-248.
Rognes K.The taxonomy of the Pollenia rudis species-group in the Holarctic Region (Diptera:Calliphoridae).Systematic Entomology,1987,12(4):475-502.
Shi YX,Xiang DP,Li ZH,Hong Y,Li XB,Xing LQ,Guo BX,Huang JC,Zheng K.COⅠ sequences analysis and molecular identification for five species of mosquitoes from Guangdong Port.Chinese Frontier Health Quarantine,2008,31(2):103-107.[师永霞,相大鹏,李祖海,洪烨,李小波,幸芦琴,郭波旋,黄吉城,郑夔.广东国境口岸不同蚊种COⅠ序列分析和分子鉴定方法.中国国境卫生检疫杂志,2008,31(2):103-107.]
Wei XY,Qiu DY,Yue QY,Liu DX.DNA barcoding and morphology identification of Pollenia rudis-a non-recorded fly species in China.Chinese Journal of Vector Biology and Control,2016,27(4):326-329.[魏晓雅,邱德义,岳巧云,刘德星.粗野粉蝇的DNA条形码和形态鉴定.中国媒介生物学及控制杂志,2016,27(4):326-329.]
Wu W,Wang FR,Xia DF.Pollenia pediculata Macquart intercepted from exotic empty containers (Diptera:Calliphoridae).Acta Parasitology et Medica Entomologica Sinica,2009,16(2):126-128.[吴薇,王芙蓉,夏德峰.入境空箱中截获的伪粗野粉蝇(双翅目:丽蝇科).寄生虫与医学昆虫学报,2009,16(2):126-128.]
Yang QG,Cai J,Song GR,Deng YH,Zhou L,He J,Hu SS,Nei GA,Sun LX.Morphology and DNA barcoding identification of Pollenia Labialis intercepted at China Port.Acta Parasitology et Medica Entomologica Sinica,2017,(3):169-173.[杨庆贵,蔡佳,宋桂荣,邓耀华,周磊,何江,胡双双,聂国嫒,孙立新.中国口岸首次截获长阳粉蝇的形态及DNA条形码鉴定.寄生虫与医学昆虫学报,2017,(3):169-173.]
Yue QY,Qiu DY,Huang YW,Hou J,Liu GX,Hu LF.Application of DNA barcoding techniques to the species identification of unknown Diptera pupa.Chinese Frontier Health Quarantine,2011,34(5):343-347.[岳巧云,邱德义,黄艺文,侯捷,刘国雄,胡龙飞.应用DNA条形码技术鉴定未知双翅目蛹.中国国境卫生检疫杂志.2011,34(5):343-347.]