洛阳地区常见嗜尸性丽蝇的COI序列分析
|
摘要
目的应用多态性较高的线粒体细胞色素C氧化酶辅酶I(CO I)对洛阳地区常见丽蝇科嗜尸性蝇类大头金蝇和丝光绿蝇进行序列分析,以期为嗜尸性蝇类的分子鉴定提供依据。方法收集洛阳地区常见大头金蝇和丝光绿蝇标本8只,经形态学分类鉴定后,Chelex-100法提取样本DNA,扩增目的片段后测序,使用相应软件对所得序列分析,建立种内及种间进化分歧率,并构建系统发育树。结果 8个样本扩增获得CO I的长度为647 bp,CO I种间平均进化分歧率0.116,种间范围0.114~0.117,种内范围0~0.002。结论 CO I基因可以较好区分所涉及的蝇种,获得的基因序列是蝇类基因库的有益补充。
Objective To analyze the common Calliphoridae flies from Luoyang by high polymorphisms of mt DNA CO I gene for identifying the species of necrophagous flies. Methods Eight flies including C.Megacephala and L. sericata were collected and classified by entomologists with traditional morphological characteristics. The DNA of flies was extracted with Chelex-100 method. The fragments of CO I were amplified and sequenced. All the sequences were analyzed by related software packages for nucleotide composition,genetic distance computation and phylogenetic tree construction. Results The result of amplification with 8 samples showed that length of the obtained CO I sequences were 647 bp. The interspecific difference in CO I ranged from 0. 114 to 0. 117 with an average of 0. 116,while the intraspecific difference was 0 to 0. 002. Conclusion The results indicated that congeneric species could be separated by CO I gene,which could be a supplement of necrophagous flie gene pool.
Objective To analyze the common Calliphoridae flies from Luoyang by high polymorphisms of mt DNA CO I gene for identifying the species of necrophagous flies. Methods Eight flies including C.Megacephala and L. sericata were collected and classified by entomologists with traditional morphological characteristics. The DNA of flies was extracted with Chelex-100 method. The fragments of CO I were amplified and sequenced. All the sequences were analyzed by related software packages for nucleotide composition,genetic distance computation and phylogenetic tree construction. Results The result of amplification with 8 samples showed that length of the obtained CO I sequences were 647 bp. The interspecific difference in CO I ranged from 0. 114 to 0. 117 with an average of 0. 116,while the intraspecific difference was 0 to 0. 002. Conclusion The results indicated that congeneric species could be separated by CO I gene,which could be a supplement of necrophagous flie gene pool.
引文
[1]Archer MS,Wallman JF.Context effects in forensic entomology and use of sequential unmasking in casework[J].J Forensic Sci,2016,61(5):1270-1277.
[2]Samerjai C,Sanit S,Sukontason K,et al.Morphology of immature stages of flesh flies,Boettcherisca nathani,and Lioproctia pattoni,(Diptera:Sarcophagidae)[J].Acta Trop,2016,163:109-120.
[3]石坚,郭亚东,匡栩源,等.应用16S r DNA短序列片段鉴定常见嗜尸性丽蝇[J].法医学杂志,2012,28(4):281-286.
[4]Brown K,Thorne AW,Harvey M.Calliphora vicina(Diptera:Calliphoridae)pupae:a timeline of external morphological development and a new age and PMI estimation tool[J].Int J Legal Med,2015,129(4):835-850.
[5]Nelson LA,Lambkin CL,Batterham P,et al.Beyond barcoding:a mitochondrial genomics approach to molecular phylogenetics and diagnostics of blowflies(Diptera:Calliphoridae)[J].Gene,2012,511(2):131-142.
[6]Harvey ML,Gaudieri S,Villet MH,et al.A global study of forensically significant calliphorids:implications for identification[J].Forensic Sci Int,2008,177(1):66-76.
[7]刘钦来.嗜尸性丽蝇分子标记的检测及发育规律的研究[D].长沙:中南大学,2013.
[8]吕宙,唐瑞,杨永强,等.嗜尸性昆虫群落演替及其法医学应用研究进展[J].中国法医学杂志,2015,30(2):166-170.
[9]Folmer O,Black M,Hoeh W,et al.DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates[J].Mol Mar Biol Biotechnol,1994,3(5):294-299.
[10]Kumar S,Stecher G,Tamura K.MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Mol Biol Evol,2016,33(7):1870-1874.
[11]Amendt J,Krettek R,Zehner R.Forensic entomology[J].Naturwissenschaften,2004,91(2):51-65.
[12]杨静波,姜建军,王江峰.mt DNA COI和ND5基因用于鉴别常见嗜尸性蝇类[J].中国法医学杂志,2012,27(3):201-204.
[13]蔡继峰.苍蝇群落演替及Mt DNA分子标记的法医学研究[D].成都:四川大学,2005.
[14]李进华,蔡继峰,郭亚东,等.利用COI序列序列鉴定常见嗜尸性蝇种[J].中国法医杂志,2011,26(6):443-446.
[15]蔡继峰,廖玍,刘敏等.嗜尸性苍蝇mt DNA中COI和COII基因序列检测及法医学应用[J].四川大学学报(医学版),2005,36(3):393-396.
[2]Samerjai C,Sanit S,Sukontason K,et al.Morphology of immature stages of flesh flies,Boettcherisca nathani,and Lioproctia pattoni,(Diptera:Sarcophagidae)[J].Acta Trop,2016,163:109-120.
[3]石坚,郭亚东,匡栩源,等.应用16S r DNA短序列片段鉴定常见嗜尸性丽蝇[J].法医学杂志,2012,28(4):281-286.
[4]Brown K,Thorne AW,Harvey M.Calliphora vicina(Diptera:Calliphoridae)pupae:a timeline of external morphological development and a new age and PMI estimation tool[J].Int J Legal Med,2015,129(4):835-850.
[5]Nelson LA,Lambkin CL,Batterham P,et al.Beyond barcoding:a mitochondrial genomics approach to molecular phylogenetics and diagnostics of blowflies(Diptera:Calliphoridae)[J].Gene,2012,511(2):131-142.
[6]Harvey ML,Gaudieri S,Villet MH,et al.A global study of forensically significant calliphorids:implications for identification[J].Forensic Sci Int,2008,177(1):66-76.
[7]刘钦来.嗜尸性丽蝇分子标记的检测及发育规律的研究[D].长沙:中南大学,2013.
[8]吕宙,唐瑞,杨永强,等.嗜尸性昆虫群落演替及其法医学应用研究进展[J].中国法医学杂志,2015,30(2):166-170.
[9]Folmer O,Black M,Hoeh W,et al.DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates[J].Mol Mar Biol Biotechnol,1994,3(5):294-299.
[10]Kumar S,Stecher G,Tamura K.MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Mol Biol Evol,2016,33(7):1870-1874.
[11]Amendt J,Krettek R,Zehner R.Forensic entomology[J].Naturwissenschaften,2004,91(2):51-65.
[12]杨静波,姜建军,王江峰.mt DNA COI和ND5基因用于鉴别常见嗜尸性蝇类[J].中国法医学杂志,2012,27(3):201-204.
[13]蔡继峰.苍蝇群落演替及Mt DNA分子标记的法医学研究[D].成都:四川大学,2005.
[14]李进华,蔡继峰,郭亚东,等.利用COI序列序列鉴定常见嗜尸性蝇种[J].中国法医杂志,2011,26(6):443-446.
[15]蔡继峰,廖玍,刘敏等.嗜尸性苍蝇mt DNA中COI和COII基因序列检测及法医学应用[J].四川大学学报(医学版),2005,36(3):393-396.