中国部分地区黑胸大蠊遗传多样性与系统发育地理结构研究
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摘要
为了探讨中国部分地区黑胸大蠊(Periplaneta fuliginosa)遗传多样性与系统发育地理结构,采用PCR产物直接测序法测定了9个地理种群88个黑胸大蠊样本的线粒体COI基因长度为658 bp的部分序列片段,开展了黑胸大蠊种群间的遗传多样性、遗传分化、基因流和系统发育地理结构分析。结果显示:黑胸大蠊COI基因共检测到11个变异位点和7个单倍型(Hap1-Hap7),其中单倍型Hap3为9个地理种群所共享;总种群的单倍型多样度(Hd)和核苷酸多样度(Pi)分别为0. 327和0. 0017,种群的中性检验Tajima's D值为-1. 2743,结果不显著,说明黑胸大蠊在较近的历史上未经历群体扩张;总种群的遗传分化系数Gst为0. 1060;基因流Nm为4. 22,表明种群间存在较强的基因交流。系统发育树和单倍型网络图分析发现各地理种群的所有单倍型并没有单独聚在一起,同来自其他地理区域种群的单倍型相隔开; AMOVA分子变异分析显示,其遗传分化主要来自种群内部,种群间未发生明显的遗传分化; Mantel分析表明,遗传距离与地理距离间无显著相关性(R=0. 0038,P=0. 5670)。上述结果表明黑胸大蠊的遗传多样性水平相对较低,且种群间没有显著的系统发育地理结构。
To investigate genetic diversity and phylogeographic structure of Periplaneta fuliginosa in some parts of China,this study applied PCR amplification and sequencing methods to determine a 658 bp fragment of partial COI gene sequences of 88 P. fuliginosa samples from 9 geographic populations. The genetic diversity,genetic differentiation,gene flow and phylogeographic structure were analyzed. The results showed that 11 variable sites and 7 haplotypes( Hap1-Hap7) were detected among all COI sequences of 9 P. fuliginosa geographic populations. Among 7 haplotypes,the haplotype Hap3 was shared by all the 9 populations. The total haplotype diversity( Hd) and nucleotide diversity( Pi) of all populations were 0. 327 and 0. 0017,respectively. The neutrality test Tajima's D was-1. 2743,indicating no sign of population expansion in recent history. The total Gst was 0. 1060. The Nm was 4. 22,showing evidence for extensive gene flow among populations. The phylogenetic tree and haplotype network analyses showed that haplotypes from different geographical regions did not cluster together and separate each region from other regions. The molecular variance analysis( AMOVA) demonstrated that genetic differentiation mainly occurred within populations,and there was no significant genetic differentiation among populations. Furthermore,a Mantel test detected no obvious correlation( R = 0. 0038,P = 0. 5670) between genetic distance and geographic distance among P. fuliginosa populations. The results indicated that genetic diversity of P. fuliginosa was relatively low and there was no phylogeographic structuring among P. fuliginosa populations.
To investigate genetic diversity and phylogeographic structure of Periplaneta fuliginosa in some parts of China,this study applied PCR amplification and sequencing methods to determine a 658 bp fragment of partial COI gene sequences of 88 P. fuliginosa samples from 9 geographic populations. The genetic diversity,genetic differentiation,gene flow and phylogeographic structure were analyzed. The results showed that 11 variable sites and 7 haplotypes( Hap1-Hap7) were detected among all COI sequences of 9 P. fuliginosa geographic populations. Among 7 haplotypes,the haplotype Hap3 was shared by all the 9 populations. The total haplotype diversity( Hd) and nucleotide diversity( Pi) of all populations were 0. 327 and 0. 0017,respectively. The neutrality test Tajima's D was-1. 2743,indicating no sign of population expansion in recent history. The total Gst was 0. 1060. The Nm was 4. 22,showing evidence for extensive gene flow among populations. The phylogenetic tree and haplotype network analyses showed that haplotypes from different geographical regions did not cluster together and separate each region from other regions. The molecular variance analysis( AMOVA) demonstrated that genetic differentiation mainly occurred within populations,and there was no significant genetic differentiation among populations. Furthermore,a Mantel test detected no obvious correlation( R = 0. 0038,P = 0. 5670) between genetic distance and geographic distance among P. fuliginosa populations. The results indicated that genetic diversity of P. fuliginosa was relatively low and there was no phylogeographic structuring among P. fuliginosa populations.
引文
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[16]柳建发,汪世平,蒋雯雯,等.黑胸大蠊的发育生物学研究进展[J].宁波大学学报(理工版),2012,25(1):125-127.
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[18]陈壮志,何苗,张成桂,等.基于COI及Cytb基因的6种蜚蠊分子系统关系[J].广州化工,2016,11(10):48-51.
[19]陈壮志,杨志斌,李贵轲,等.基于COI和18S rRNA基因对蜚蠊物种进行分子鉴定[J].广州化工,2016,44(13):104-107.
[20]CHO S Y,SUH K I,BAE Y J. DNA barcode library and its efficacy for identifying food-associated insect pests in Korea[J]. Entomological Research,2013,43(5):253-261.
[21]FOLMER O,BLACK M,HOEH W,et al. DNA primers for amplification of mitochondrial cytochrome coxidase subunit I from diverse metazoan invertebrates[J]. Molecular Marine Biology&Biotechnology,1994,3(5):294-299.
[22]LEIGH J W,BRYANT D. Popart:full-feature software for haplotype network construction[J]. Methods in Ecology&Evolution,2015,6(9):1110-1116.
[23]HEBERT P D N,CYWINSKA A,BALL S L,et al. Biological identifications through DNA barcodes[J]. Proceedings of the Royal Society B,2003,270(1512):313-321.
[24]JINFU W,CHAOHUI H. Molecular divergence of the mitochondrial cytochrome oxidase II gene in three mosquitoes[J]. Journal of the American Mosquito Control Association,2002,18(4):301-306.
[25]XIAO Y,ZHANG Y,GAO T,et al. Genetic diversity in the mt DNA control region and population structure in the small yellow croaker Larimichthys polyactis[J]. Environmental Biology of Fishes,2009,85(4):303-314.
[26]YAMAGUCHI K,NAKAJIMA M,TANIGUCHI N. Loss of genetic variation and increased population differentiation in geographically peripheral populations of Japanese char Salvelinus leucomaenis[J].Aquaculture,2010,308(1):S20-S27.
[27]魏中华,郭明霞,徐娟,等.居室黑胸大蠊食性和食物范围研究[J].安徽农业科学,2015,43(2):72-73.
[28]李春选,马恩波,郑先云.中国4种蝗虫不同种群的遗传分化[J].遗传学报,2003,30(3):234-244.
[29]ROUSSET F. Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance[J]. Genetics,1997,145(4):1219-1228.
[30]MILLAR C I,LIBBY W J,FALK D A,et al. Strategies for conserving clinal,ecotypic,and disjunct population diversity in widespread species[M]. New Oxford:Oxford University Press,1991:149-170.
[31]WHITLOCK M C,MCCAULEY D E. Indirect measures of gene flow and migration:Fst≠1/(4Nm+1). Heredity[J]. Heredity,1999,82(2):117-125.
[2]BAUMHOLTZ M A,PARISH L C,WITKOWSKI J A,et al. The medical importance of cockroaches[J]. International Journal of Dermatology,1997,36(2):90-96.
[3]SAICHUA P,PINMAI K,SOMRITHIPOL S,et al. Isolation of medically important fungi from cockroaches trapped at Thammasat Chalermprakiat Hospital[J]. Thammasat Medical Journal,2008,8(3):345-351.
[4]SHAHRAKI G H,PARHIZKAR S,NEJAD A R S. Cockroach infestation and factors affecting the estimation of cockroach population in urban communities[J]. International Journal of Zoology,2013,2013(2):1-6.
[5]TATFENG Y M,USUANLELE M U,ORUKPE A,et al. Mechanical transmission of pathogenic organisms:the role of cockroaches[J]. Journal of Vector Borne Diseases,2005,42(4):129-134.
[6]吴仕筠,米长钟,郭靖,等.黑胸大蠊的形态学及显微结构研究[J].中国民族民间医药,2008,17(8):14-16.
[7]徐绍锐,汪世平,吴仕筠,等.黑胸大蠊不同发育阶段可溶性抗原特征分析[J].中国人兽共患病学报,2004,20(4):291-294.
[8]PESOLE G,GISSI C,DE CHIRICO A. et al. Nucleotide substitution rate of mammalian mitochondrial genomes[J]. Journal of Molecular Evolution,1999,48(4):427-434.
[9]ROKAS A,LADOUKAKIS E,ZOUROS E. Animal mitochondrial DNA recombination revisited[J]. Trends in Ecology&Evolution,2003,18(8):411-417.
[10]PAKENDORF B,STONEKING M. Mitochondrial DNA and human evolution[J]. Annual Review of Genomics and Human Genetics,2005,6:165-183.
[11]杨小强.基于线粒体COI序列的中国东南地区马六甲肉食螨的系统地理结构研究[D].南昌:南昌大学,2013.
[12]任秋平.基于匿名核基因和线粒体基因初步探讨中国南方地区双纹小蠊的种群结构[D].南京:南京师范大学,2013.
[13]王红,徐忠新,韩岚岚,等.基于线粒体COI基因序列的大豆食心虫中国东北地理种群遗传多样性分析[J].昆虫学报,2014,57(9):1051-1060.
[14]徐绍锐.黑胸大蠊生物学特性及药用价值的研究[D].长沙:中南大学,2005.
[15]徐绍锐,汪世平,吴仕筠,等.黑胸大蠊室内繁殖、发育的生物学特性研究[J].中国人兽共患病学报,2006,22(10):913-917.
[16]柳建发,汪世平,蒋雯雯,等.黑胸大蠊的发育生物学研究进展[J].宁波大学学报(理工版),2012,25(1):125-127.
[17]黄利,吴仕筠.黑胸大蠊水提取物对H22荷瘤小鼠肿瘤的抑制作用[J].吉首大学学报(自然版),2016,37(3):85-89.
[18]陈壮志,何苗,张成桂,等.基于COI及Cytb基因的6种蜚蠊分子系统关系[J].广州化工,2016,11(10):48-51.
[19]陈壮志,杨志斌,李贵轲,等.基于COI和18S rRNA基因对蜚蠊物种进行分子鉴定[J].广州化工,2016,44(13):104-107.
[20]CHO S Y,SUH K I,BAE Y J. DNA barcode library and its efficacy for identifying food-associated insect pests in Korea[J]. Entomological Research,2013,43(5):253-261.
[21]FOLMER O,BLACK M,HOEH W,et al. DNA primers for amplification of mitochondrial cytochrome coxidase subunit I from diverse metazoan invertebrates[J]. Molecular Marine Biology&Biotechnology,1994,3(5):294-299.
[22]LEIGH J W,BRYANT D. Popart:full-feature software for haplotype network construction[J]. Methods in Ecology&Evolution,2015,6(9):1110-1116.
[23]HEBERT P D N,CYWINSKA A,BALL S L,et al. Biological identifications through DNA barcodes[J]. Proceedings of the Royal Society B,2003,270(1512):313-321.
[24]JINFU W,CHAOHUI H. Molecular divergence of the mitochondrial cytochrome oxidase II gene in three mosquitoes[J]. Journal of the American Mosquito Control Association,2002,18(4):301-306.
[25]XIAO Y,ZHANG Y,GAO T,et al. Genetic diversity in the mt DNA control region and population structure in the small yellow croaker Larimichthys polyactis[J]. Environmental Biology of Fishes,2009,85(4):303-314.
[26]YAMAGUCHI K,NAKAJIMA M,TANIGUCHI N. Loss of genetic variation and increased population differentiation in geographically peripheral populations of Japanese char Salvelinus leucomaenis[J].Aquaculture,2010,308(1):S20-S27.
[27]魏中华,郭明霞,徐娟,等.居室黑胸大蠊食性和食物范围研究[J].安徽农业科学,2015,43(2):72-73.
[28]李春选,马恩波,郑先云.中国4种蝗虫不同种群的遗传分化[J].遗传学报,2003,30(3):234-244.
[29]ROUSSET F. Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance[J]. Genetics,1997,145(4):1219-1228.
[30]MILLAR C I,LIBBY W J,FALK D A,et al. Strategies for conserving clinal,ecotypic,and disjunct population diversity in widespread species[M]. New Oxford:Oxford University Press,1991:149-170.
[31]WHITLOCK M C,MCCAULEY D E. Indirect measures of gene flow and migration:Fst≠1/(4Nm+1). Heredity[J]. Heredity,1999,82(2):117-125.
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