基于DNA条形码丽蝇科49种常见种的分类鉴定

doi:10.11853/j.issn.1003.8280.2017.04.005

摘要/Abstract

摘要： 目的建立丽蝇科49种常见种的DNA条形码鉴定方法，通过获得其DNA条形码数据为丽蝇科种类鉴定提供技术支持和参比数据，并解决个别雌虫、残缺个体和幼虫等无法准确鉴定的难题。方法对2012-2015年采自全国各地的丽蝇科49种166份标本进行形态学鉴定，并获取相应种类雌雄个体的DNA条形码数据，与已有序列共同构建系统进化树，验证DNA条形码鉴定与形态学鉴定结果的一致性。结果丽蝇科49种的166条序列均提交至GenBank数据库，其中28种丽蝇DNA条形码序列为全球范围内首次获得。丽蝇科同种不同个体间差异<1%，不同种间的DNA条形码差异度随形态学差异的增大而增高。丽蝇科常见种类DNA条形码鉴定结果与形态学鉴定结果一致。结论 DNA条形码是传统分类学的有力补充，特别是针对昆虫的残体、非成虫态及雌性个体均可鉴定，可提高物种鉴定的准确度，对国境口岸准确快速地鉴定媒介生物种类有重要意义。

关键词: 丽蝇科, DNA条形码, 分类鉴定

Abstract: Objective To establish the DNA barcoding identification techniques for 49 common fly species in Calliphoridae, and provide references data for the identification of these species with DNA barcodes, as to solve the difficulties in the accurate identification of the females, individuals with lost morphological features, and larvae of these species. Methods Totally 166 individuals in Calliphoridae were identified with the morphological characters which were collected from different areas in China during 2012 to 2015, DNA barcodes of the males and females were obtained, an N-J tree was constructed in combination with published data. Consistency of the identification results by morphology and DNA barcodes were tested as well. Results All the 166 sequences of these 49 species were submitted to the public data base, DNA barcodes sequences of 28 species were obtained for the first time. The differences of the intra-species was less than 1%, and the inter-species DNA barcode differences increasing with the morphology differences, and the identification results based on DNA barcodes were consistent with that based on morphology. Conclusion The application of DNA barcodes were strong supplemental to the morphology identification of the species, especially to the incomplete, non-adult and female individuals, and can increase accuracy of identification, and it is very important for the rapid and accurate species identification of the medical vectors at the ports.

Key words: Calliphoridae, DNA barcodes, Species identification

中图分类号:

R384.2

邱德义, 廖俊蕾, 魏晓雅, 岳巧云. 基于DNA条形码丽蝇科49种常见种的分类鉴定[J]. 中国媒介生物学及控制杂志, 2017, 28(4): 322-331.

QIU De-yi, LIAO Jun-lei, WEI Xiao-ya, YUE Qiao-yun. Identification of 49 common fly species in Calliphoridae based on DNA barcodes[J]. Chines Journal of Vector Biology and Control, 2017, 28(4): 322-331.

参考文献

[1] 范滋德,陈之梓,方建明,等. 中国动物志. 昆虫纲. 第6卷. 双翅目:丽蝇科[M]. 北京:科学出版社,1997:120-125.
[2] 吴薇,夏德峰,郑炜,等. 浙江省丽蝇科区系研究[J]. 广东农业科学,2014,41(9):88-90.
[3] Crosskey RW, Lane RP. House-flies, blow-flies and their allies (Calyptrate:Diptera)[M]//Lane RP, Crosskey RW. Medical Insects and Arachnids. London:Chapman and Hall,1993:403-428.
[4] Hall M, Wall R. Myiasis of humans and domestic animals[J]. Adv Parasitol,1995,35:257-334.
[5] Sherman RA,Hall MJR,Thomas S. Medicinal maggots:an ancient remedy for some contemporary afflictions[J]. Annu Rev Entomol,2000,45(1):55-81.
[6] Amendt J, Krettek R, Zehner R. Forensic entomology[J]. Naturwissenschaften,2004,91(2):51-65.
[7] Rognes K. Blowflies (Diptera:Calliphoridae) of Fennoscandia and Denmark[M]. New York:Brill Academic Publishers, 1991,24:1-272.
[8] Stevens J, Wall R. Species, sub-species and hybrid populations of the blowflies Lucilia cuprina and Lucilia sericata (Diptera:Calliphoridae)[J]. Proc Roy Biol Soc B,1996,263(1375):1335-1341.
[9] Harvey ML,Dadour IR,Gaudieri S. Mitochondrial DNA cytochrome oxidase I gene:potential for distinction between immature stages of some forensically important fly species (Diptera) in western Australia[J]. Forensic Sci Int,2003,131(2/3):134-139.
[10] Chen WY, Hung TH, Shiao SF. Molecular identification of forensically important blow fly species (Diptera:Calliphoridae) in Taiwan[J]. J Med Entomol,2004,41(1):47-57.
[11] Saigusa K, Takamiya M, Aoki Y. Species identification of the forensically important flies in Iwate prefecture, Japan based on mitochondrial cytochrome oxidase gene subunitⅠ (COⅠ) sequences[J]. Legal Med,2005,7(3):175-178.
[12] Wells JD, Stevens JR. Application of DNA-based methods in forensic entomology[J]. Annu Rev Entomol,2008,53(1):103-120.
[13] Hebert PDN,Cywinska A,Ball SL,et al. Biological identifications through DNA barcodes[J]. Proc Biol Sci,2003,270(1512):313-321.
[14] Hebert PDN, Ratnasingham S, deWaard JR. Barcoding animal life:cytochrome c oxidase subunit 1 divergences among closely related species[J]. Pro Biol Sci,2003,270 Suppl 1:S96-99.
[15] Hebert PDN, Penton EH, Burns JM, et al. Ten species in one:DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator[J]. Proc Natl Acad Sci USA,2004,101(41):14812-14817.
[16] Hajibabaei M, Janzen DH, Burn JM, et al. DNA barcodes distinguish species of tropical Lepidoptera[J]. Proc Natl Acad Sci USA,2006,103(4):968-971.
[17] Ratnasingham S, Hebert PDN. BOLD:the barcode of life data system (http://www.barcodinglife.org)[J]. Mol Ecol Res,2007,7(3):355-364.
[18] Stoeckle M. Taxonomy, DNA, and the barcode of life[J]. Bio Sci,2003,53(9):796-797.
[19] GilArriortua M, Bordas MIS, Cainé LM, et al. Cytochrome b as a useful tool for the identification of blowflies of forensic interest (Diptera:Calliphoridae)[J]. Forensic Sci Int,2013,228(1/3):132-136.
[20] Rolo EA, Oliveira AR, Dourado CG, et al. Identification of sarcosaprophagous Diptera species through DNA barcoding in wildlife forensics[J]. Forensic Sci Int,2013,228(1/3):160-164.
[21] Will KW, Rubinoff D. Myth of the molecule:DNA barcodes for species cannot replace morphology for identification and classification[J]. Cladistics,2004,20(1):47-55.
[22] Monaghan MT, Balke M, Gregory TR, et al. DNA-based species delineation in tropical beetles using mitochondrial and nuclear markers[J]. Philos Trans R Soc Lond B Biol Sci,2005, 360(1462):1925-1933.
[23] Foley DH, Wilkerson RC, Cooper RD, et al. A molecular phylogeny of Anopheles annulipes (Diptera:Culicidae) sensu lato:the most species-rich anopheline complex[J]. Mol Phylogenet Evol,2007,43(1):287-297.
[24] Blacket MJ, Semeraro L, Malipatil MB. Barcoding Queensland fruit flies (Bactrocera tryoni):impediments and improvements[J]. Mol Ecol Res,2012,12(3):428-436.
[25] Nelson LA, Wallman JF, Dowton M. Using COⅠ barcodes to identify forensically and medically important blowflies[J]. Med Vet Entomol,2007,21(1):44-52.
[26] Wallman JF, Donnellan SC. The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera:Calliphoridae) in southeastern Australia[J]. Forensic Sci Int,2001,120(1/2):60-67.
[27] Wells JD, Wall R, Stevens JR. Phylogenetic analysis of forensically important Lucilia flies based on cytochrome oxidaseⅠ sequence:a cautionary tale for forensic species determination[J]. Int J Legal Med,2007,121(3):229-233.
[28] Meiklejohn KA, Wallman JF, Dowton M. DNA-based identification of forensically important Australian Sarcophagidae (Diptera)[J]. Int J Legal Med,2011,125(1):27-32.
[29] Smith MA, Woodley NE, Janzen DH, et al. DNA barcodes reveal cryptic host-specificity within the presumed polyphagous members of a genus of parasitoid flies (Diptera:Tachinidae)[J]. Proc Natl Acad Sci USA,2006,103(10):3657-3662.
[30] 岳巧云,邱德义,黄艺文,等. DNA条形码技术在未知昆虫幼虫种类鉴定中的应用[J]. 中国卫生检验杂志,2011,21(3):615-617.
[31] 刘德星,聂维忠,邱德义,等. 应用DNA条形码技术快速鉴定入境船舶上截获的昆虫蛹[J]. 检验检疫学刊,2014,24(5):53-57.
[32] Besansky NJ, Severson DW, Ferdig MT. DNA barcoding of parasites and invertebrate disease vectors:what you don't know can hurt you[J]. Trends Parasitol,2003,19(12):545-546.
[33] 范滋德. 中国常见蝇类检索表[M]. 2版. 北京:科技出版社, 1992:457-580.
[34] 薛万琦,赵建铭. 中国蝇类[M]. 沈阳:辽宁科学技术出版社, 1998:1366-1506.
[35] Yue QY, Wu KL, Qiu DY, et al. A formal re-description of the cockroach Hebardina concinna anchored on DNA barcodes confirms wing polymorphism and identifies morphological characters for field identification[J]. PLoS One,2014,9(9):e106789. DOI:10.1371/journal.pone.0106789.
[36] Folmer O,Black M,Hoeh W,et al. DNA primers for amplification of mitochondrial cytochrome c oxidase subunitⅠ from diverse metazoan invertebrates[J]. Mol Mar Biol Biotechnol,1994,3(5):294-299.
[37] Kumar S, Nei M, Dudley J, et al. MEGA:a biologist-centric software for evolutionary analysis of DNA and protein sequences[J]. Brief Bioinform,2008,9(4):299-306.
[38] GilArriortua M, Bordas MIS, K-hnemann S, et al. Molecular differentiation of central European blowfly species (Diptera:Calliphoridae) using mitochondrial and nuclear genetic markers[J]. Forensic Sci Int,2014,242:274-282.
[39] Renaud AK, Savage J, Adamowicz SJ. DNA barcoding of northern Nearctic Muscidae (Diptera) reveals high correspondence between morphological and molecular species limits[J]. BMC Ecol,2012,12:24.
[40] Sonet G, Jordaens K, Braet Y, et al. Why is the molecular identification of the forensically important blowfly species Lucilia caesar and L. illustris (family Calliphoridae) so problematic?[J]. Forensic Sci Int,2012,223(1/3):153-159.
[41] Wells JD,Sperling FAH. DNA-based identification of forensically important Chrysomyinae (Diptera:Calliphoridae)[J]. Forensic Sci Int,2001,120(1/2):110-115.
[42] Reibe S, Schmitz J, Madea B. Molecular identification of forensically important blowfly species (Diptera:Calliphoridae) from Germany[J]. Parasitol Res,2009,106(1):257-261.