Identification of Common Sarcosaprophagous Flies in the Yangtze River Delta by COⅠ Gene

doi:10.12116/j.issn.1004-5619.2020.400611

Abstract

Abstract: Objective

To identify the common sarcosaprophagous flies in the Yangtze River Delta based on mitochondrial cytochrome c oxidase subunit Ⅰ（COⅠ） gene sequence and verify the reliability of this method.

Methods

Seven common genetically stable sarcosaprophagous flies in three families and six genera were collected from large domestic pig carcasses placed in the field and cultured in the laboratory for many generations. The whole genome DNA was extracted and the COⅠ gene fragment was amplified. The forward and reverse sequencing was followed by splicing. The base composition of the amplified fragment and the rate of interspecific evolutionary divergence were analyzed by software such as Mega 7.0.26. The phylogenetic tree of COⅠ gene sequence of common necrophagous flies in the Yangtze River Delta was established by neighbor joining （NJ） method and unweighted pair-group method with arithmetic means （UPGMA） method.

Results

The average base composition of different flies was A（30.14%）, T（38.23%）, C（15.98%）, G（15.65%）. The rate of interspecific evolutionary divergence ranged from 2.2% to 15.3%, the lowest rate was between Chrysomya megacephala and Chrysomya pinguis, the highest rate was between Muscina stabulans and Boettcherisca peregrina.

Conclusion

COⅠ gene can be used to identify the common necrophagous flies in the Yangtze River Delta.

Key words: forensic entomology, cytochrome c oxidase subunit Ⅰ, species identification, sarcosaprophagous flies, Yangtze river delta

CLC Number:

DF795.1

Wang XU, Yu WANG, Man WANG, Ying-hui WANG, Jiang-feng WANG. Identification of Common Sarcosaprophagous Flies in the Yangtze River Delta by COⅠ Gene[J]. Journal of Forensic Medicine, 2021, 37(5): 611-614.

Figures/Tables 4

References 15

1	WANG M， CHU J， WANG Y， et al. Forensic entomology application in China： Four case reports[J]. J Forensic Leg Med，2019，63：40-47. doi：10.1016/j.jflm.2019.03.001.
2	AMENDT J， KRETTEK R， ZEHNER R. Forensic entomology[J]. Sci Nat-Heidelberg，2004，91（2）：51-65. doi：10.1007/s00114-003-0493-5.
3	KAKOULI-DUARTE T， CASEY D G， BURNELL A M. Development of a diagnostic DNA probe for the fruit flies Ceratitis capitata and Ceratitis rosa （Diptera： Tephritidae） using amplified fragment-length polymorphism[J]. J Econ Entomol，2001，94（4）：989-997. doi：10.1603/0022-0493-94.4.989.
4	陈庆，白洁，刘力，等. 北京地区7种常见嗜尸性蝇类的COⅠ基因序列分析及DNA条形码的建立[J].昆虫学报，2009，52（2）：202-209. doi：10.3321/j.issn：0454-6296.2009.02.012.
	CHEN Q， BAI J， LIU L， et al. Sequential analysis of mitochondrial COⅠ gene for seven common sarcosaphagous flies （Diptera） in Beijing and the establishment of their DNA barcodes[J]. Kunchong Xuebao，2009，52（2）：202-209.
5	GUO Y， ZHA L， YAN W， et al. Identification of forensically important sarcophagid flies （Diptera： Sarcophagidae） in China based on COⅠ and period gene[J]. Int J Legal Med，2014，128（1）：221-228. doi：10.1007/s00414-013-0923-7.
6	MÁRQUEZ-ACERO A S， VIDAL-GARCIA J M， GALINDO L F B， et al. Molecular identification of six species of Calliphoridae （Diptera） with forensic interest in Bogotá， Colombia[J]. Papéis Avulsos de Zoologia，2017，57（12）：141-147. doi：10.11606/0031-1049.2017.57.12.
7	张学卫，张智婷，原占国，等. 昆虫线粒体DNA Cyt b基因研究进展[J]. 河北林果研究，2010，25（2）：173-176. doi：10.3969/j.issn.1007-4961.2010.02.017.
	ZHANG X W， ZHANG Z T， YUAN Z G， et al. Mitochondrial DNA Cyt b gene of insect[J]. Hebei Linguo Yanjiu，2010，25（2）：173-176.
8	GILARRIORTUA M， SALONA B M， KOHNEMANN S， et al. Molecular differentiation of central European blowfly species （Diptera， Calliphoridae） using mitochon
9	BHARTI M， SINGH B. DNA-based identification of forensically important blow flies （Diptera： Calliphoridae） from India[J]. J Med Entomol，2017，54（5）：1151-1156. doi：10.1093/jme/tjx084.
10	HEBERT P D， CYWINSKA A， BALL S L， et al. Biological identifications through DNA barcodes[J]. Proc Biol Sci，2003，270（1512）：313-321. doi：10.10 98/rspb.2002.2218.
11	WELLS J D， SPERLING F A. DNA-based identification of forensically important Chrysomyinae （Diptera： Calliphoridae）[J]. Forensic Sci Int，2001，120（1/2）：110-115. doi：10.1016/s0379-0738（01）00414-5.
12	NELSON L A， WALLMAN J F， DOWTON M. Using COⅠ barcodes to identify forensically and medically important blowflies[J]. Med Vet Entomol，2007，21（1）：44-52. doi：10.1111/j.1365-2915.2007.00 664.x.
13	NELSON L A， WALLMAN J F， DOWTON M. Identification of forensically important Chrysomya （Diptera： Calliphoridae） species using the second ribosomal internal transcribed spacer （ITS2）[J]. Forensic Sci Int，2008，177（2/3）：238-247. doi：10.1016/j.forsciint.2008.01.009.
14	赵琳琳，翟仙敦，郑哲，等. 洛阳地区9种嗜尸性丽蝇28S rRNA和COⅠ基因序列分析[J].法医学杂志， 2019，35（2）：181-186. doi：10.12116/j.issn.1004-5619. 2019.02.010.
	ZHAO L L， ZHAI X D， ZHENG Z， et al. Analysis of 28S rRNA and COⅠ gene sequence of nine necrophagous Calliphorid flies from Luoyang[J]. Fayixue Zazhi，2019，35（2）：181-186.
15	卓荦，毛佳雄，陈建山，等. 福建省常见尸食性蝇类的COⅠ及16S rDNA序列鉴定[J].法医学杂志，2020，36（6）：749-754. doi：10.12116/j.issn.1004-5619.2020.06.002.
	ZHUO L， MAO J X， CHEN J S， et al. COⅠ and 16S rDNA sequence identification of common necrophagous flies in Fujian province[J]. Fayixue Zazhi，2020，36（6）：749-754.

科	属	种	拉丁名
丽蝇科	金蝇属	大头金蝇	Chrysomya megacephala
		肥躯金蝇	Chrysomya pinguis
	裸金蝇属	绯颜裸金蝇	Chrysomya rufifacies
	阿丽蝇属	巨尾阿丽蝇	Chrysomya grahami
	绿蝇属	丝光绿蝇	Lucilia sericata
麻蝇科	别麻蝇属	棕尾别麻蝇	Boettcherisca peregrina
蝇科	厩腐蝇属	厩腐蝇	Muscina stabulans

科	属	种	拉丁名
丽蝇科	金蝇属	大头金蝇	Chrysomya megacephala
		肥躯金蝇	Chrysomya pinguis
	裸金蝇属	绯颜裸金蝇	Chrysomya rufifacies
	阿丽蝇属	巨尾阿丽蝇	Chrysomya grahami
	绿蝇属	丝光绿蝇	Lucilia sericata
麻蝇科	别麻蝇属	棕尾别麻蝇	Boettcherisca peregrina
蝇科	厩腐蝇属	厩腐蝇	Muscina stabulans

蝇类	大头金蝇	绯颜裸金蝇	巨尾阿丽蝇	厩腐蝇	棕尾别麻蝇	丝光绿蝇
大头金蝇	-
绯颜裸金蝇	6.5	-
巨尾阿丽蝇	8.7	9.7	-
厩腐蝇	12.4	14.0	11.5	-
棕尾别麻蝇	13.6	13.6	14.6	15.3	-
丝光绿蝇	9.2	11.5	7.2	10.8	12.6	-
肥躯金蝇	2.2	7.7	9.2	12.4	14.2	9.7

蝇类	大头金蝇	绯颜裸金蝇	巨尾阿丽蝇	厩腐蝇	棕尾别麻蝇	丝光绿蝇
大头金蝇	-
绯颜裸金蝇	6.5	-
巨尾阿丽蝇	8.7	9.7	-
厩腐蝇	12.4	14.0	11.5	-
棕尾别麻蝇	13.6	13.6	14.6	15.3	-
丝光绿蝇	9.2	11.5	7.2	10.8	12.6	-
肥躯金蝇	2.2	7.7	9.2	12.4	14.2	9.7

蝇类	A	T	C	G	A+T	C+G
大头金蝇	30.40	38.45	15.35	15.81	68.85	31.15
绯颜裸金蝇	30.55	38.16	15.65	15.65	68.71	31.29
巨尾阿丽蝇	30.85	37.99	15.96	15.20	68.84	31.16
厩腐蝇	29.03	38.45	16.72	15.81	67.48	32.52
棕尾别麻蝇	29.33	37.69	17.02	15.96	67.02	32.98
丝光绿蝇	30.55	39.06	15.05	15.35	69.61	30.39
肥躯金蝇	30.24	37.84	16.11	15.81	68.08	31.92
均值	30.14	38.23	15.98	15.65	68.37	31.63