基于线粒体Cytb基因的意大利蝗遗传多样性分析
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摘要
【目的】分析新疆荒漠半荒漠草原意大利蝗Calliptamus italicus不同地理种群的遗传多样性,探讨新疆意大利蝗持续严重发生的分子生物学机制。【方法】测定并获得意大利蝗10个不同地理种群的100条Cytb基因全长序列(1 140 bp),利用DnaSP 5.0和Arlequin 3.5软件分析种群遗传多样性、遗传结构、遗传分化与分子变异,并构建了最大似然法系统发育树及单倍型中介网络图。【结果】在100个序列样本中,共检测出54个单倍型。总群体单倍型多样度H_d为0.890,各地理种群单倍型多样度范围介于0.818-1.000,总群体固定系数F_(st)、遗传分化系数分别为0.017 8、0.018 9,总基因流N_m为13.84,种群间遗传差异(3.31%)远小于种群内的遗传变异(96.69%),结果表明,意大利蝗群体稳定,历史上未出现群体扩张,地理距离可能不是影响种群间遗传距离的重要因素。【结论】新疆意大利蝗不同地理种群间基因交流频繁,通过种群内遗传变异提高适应外界环境能力,可能是其持续严重发生的重要原因之一。
[Objectives] To analyze the genetic variation among different geographic populations of Calliptamus italicus in desert and semi-desert steppe in Xinjiang, and to reveal the molecular and biological mechanisms underlying recent persistent and serious outbreaks of this pest. [Methods] Specimens of C. italicus were collected from 10 different geographic populations to obtain a total of 100 full-length sequences of the mitochondrial cytochrome b(Cytb) gene(1 140 bp). We then used DnaSP 5.0 and Arlequin 3.5 software to clarify the genetic structure, differentiation, and molecular variances, within and among populations. A Maximum Likelihood phylogenetic tree and a diagram of the haplotype network of Cytb were constructed. [Results] We detected 54 haplotypes among the 100 full-length sequences. The total haplotype diversity index(H_d) was 0.890 and haplotype diversity among the different geographical populations ranged from 0.818 to 1.000. The total fixed coefficient(F_(st)), total genetic differentiation coefficient(G_(st)), and total gene flow(N_m) were 0.0178, 0.0189 and 13.84,respectively. In addition, the genetic variation between populations(3.31%) was much less than that within each population.These results indicate that the C. italicus populations investigated in this study are stable in size and have not undergone recent population expansion. Geographic distance may not, therefore, be an important factor affecting the genetic distance between different populations. [Conclusion] Gene exchange occurs frequently between different geographical populations of C.italicus in Xinjiang, increasing the genetic variation within populations and thereby enhancing their ability to adapt to different environmental conditions. This situation may be related to recent persistent and serious outbreaks of C. italicus in Xinjiang.
[Objectives] To analyze the genetic variation among different geographic populations of Calliptamus italicus in desert and semi-desert steppe in Xinjiang, and to reveal the molecular and biological mechanisms underlying recent persistent and serious outbreaks of this pest. [Methods] Specimens of C. italicus were collected from 10 different geographic populations to obtain a total of 100 full-length sequences of the mitochondrial cytochrome b(Cytb) gene(1 140 bp). We then used DnaSP 5.0 and Arlequin 3.5 software to clarify the genetic structure, differentiation, and molecular variances, within and among populations. A Maximum Likelihood phylogenetic tree and a diagram of the haplotype network of Cytb were constructed. [Results] We detected 54 haplotypes among the 100 full-length sequences. The total haplotype diversity index(H_d) was 0.890 and haplotype diversity among the different geographical populations ranged from 0.818 to 1.000. The total fixed coefficient(F_(st)), total genetic differentiation coefficient(G_(st)), and total gene flow(N_m) were 0.0178, 0.0189 and 13.84,respectively. In addition, the genetic variation between populations(3.31%) was much less than that within each population.These results indicate that the C. italicus populations investigated in this study are stable in size and have not undergone recent population expansion. Geographic distance may not, therefore, be an important factor affecting the genetic distance between different populations. [Conclusion] Gene exchange occurs frequently between different geographical populations of C.italicus in Xinjiang, increasing the genetic variation within populations and thereby enhancing their ability to adapt to different environmental conditions. This situation may be related to recent persistent and serious outbreaks of C. italicus in Xinjiang.
引文
Azhbenov VK,Baybussenov KS,Sarbaev AT,Harizanova VB,2015.Preventive approach of phytosanitary control of locust pests in Kazakhstan and adjacent areas.International Conference on Agricultural,Ecological and Medical Sciences.33-37.
Boivin T,Bouvier JC,Beslay D,Sauphanor B,2004.Variability in diapause propensity within populations of a temperate insect species:interactions between insecticide resistance genes and photoperiodism.Biological Journal of the Linnean Society,83(3):341-351.
Chen QL,Tang XS,Yao WJ,Lu SQ,2009.Bioinformatics analysis of the complete sequences of cytochrome b of Takydromus sylvaticus and modeling the tertiary structure of encoded protein.International Journal of Biological Sciences,5(6):596-602.
Chen YL,Liu JP,1979.The Control of Locust in Xinjiang.Urumqi:Volksverlag Xinjiang.9-10.[陈永林,刘举鹏,1979.新疆蝗虫及其防治.乌鲁木齐:新疆人民出版社.9-10.]
Chen YL,2000.Resurgent control and ecological management of locust.Bulletin of Chinese Academy of Sciences,(5):341-345.[陈永林,2000.蝗虫再猖獗的控制与生态学治理.中国科学院院刊,(5):341-345.]
Dai JX,2005.Cytochrome b gene and molecular systematology research of insects.Sichuan Journal of Zoology,24(2):222-225.[戴金霞,2005.线粒体Cytb基因与昆虫分子系统学研究.四川动物,24(2):222-225.]
Dou J,Zhang RY,Liu M,Qian X,Xiao HW,Roman J,Ji R,2017.Comparison of flight muscle and energy consumption of pre-and post-flight in Calliptamus italicus(Orthoptera:Acrididae).Pratacultural Science,34(8):1721-1726.[窦洁,张若燕,刘敏,钱雪,肖宏伟,Roman Jashenko,季荣,2017.飞行前后意大利蝗飞行肌及能源消耗比较.草业科学,34(8):1721-1726.]
Excoffier L,Lischer HEL,2010.Arlequin suite ver 3.5.A new series of programs to perform population genetics analyses under Linux and Windows.Molecular Ecology Resources,10(3):564-567.
FAO Locust Watch Release,2011.http://www.fao.org/ag/locustsCCA/en/1010/1018/1075/.
Gray MW,1989.Origin and evolution of mitochondrial DNA.Annual Review of Cell Biology,5(5):25.
Guo XZ,1993.Forage resources in the South Mountain area in Urumqi.Xinjiang Animal Husbandry,(4):23-25.[郭选政,1993.乌鲁木齐南山地区的牧草资源.新疆畜牧业,(4):23-25.]
Ha M,Han XH,2014.The current situation of the grassland locusts and the strategies for prevention and treatment in the border area of Tacheng region,Xinjiang.Journal of Agriculture,4(3):7-10.[哈玛尔,韩小虎,2014.新疆塔城地区边境草原蝗虫发生现状及防治对策.农学学报,4(3):7-10.]
Huang H,Zhu EL,2001.The serious outbreak of locusts in Kazakhstan.Word Agriculture,(6):46-47.[黄辉,朱恩林,2001.哈萨克斯坦蝗灾严重发生.世界农业,(6):46-47.]
Kimura M,1980.A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.Journal of Molecular Evolution,16(2):111-120.
Kokanova EO,2014.The current state of natural foci of swarming locusts(Orthoptera,Acrididae)in Turkmenistan.Entomological Review,94(6):826-828.
Leigh JW,Bryant D,2015.PopART:Full-feature software for haplotype network construction.Methods in Ecology and Evolution,6(9):1110-1116.
Li JB,Ren ZM,2009.Genetic diversity among Schlechtendalia chinensis individuals revealed by Cytb sequences.Journal of Fudan University(Natural Science),48(5):680-686.[李继变,任竹梅,2009.角倍蚜mtDNA Cytb基因遗传多样性分析.复旦学报(自然科学版),48(5):680-686.]
Li PX,Lin J,Mai D,Wu JG,Liu YQ,Zheng JH,2017.Effects of future climate change on potential geographic distribution of Calliptamus italicus(L.)in Xinjiang.Plant Protection,43(3):90-96.[李培先,林峻,麦迪·库尔曼,吴建国,刘永强,郑江华,2017.气候变化对新疆意大利蝗潜在分布的影响.植物保护,43(3):90-96.]
Li R,Han LL,Wang H,Zhao KJ,Mang YL,Zhang HY,Fan D,2015.Analysis of genetic differentiation among different geographic populations of Aphis glycines(Hemiptera:Aphididae)based on mtDNA COII gene sequences.Chinese Journal of Applied Entomology,52(5):1203-1214.[李冉,韩岚岚,王红,赵奎军,莽逸伦,张红玉,樊东,2015.基于线粒体COII基因的大豆蚜不同地理种群遗传分化研究.应用昆虫学报,52(5):1203-1214.]
Liu ZG,Ji T,Shen F,Liang Q,Luo YX,2015.Genetic diversity of geographic populations of Apis cerana cerana estimated by mitochondrial COII gene sequences.Journal of Environmental Entomology,37(3):567-575.[刘振国,吉挺,沈芳,梁勤,罗岳雄,2015.基于线粒体COII基因序列的中华蜜蜂地理种群的遗传多样性研究.环境昆虫学报,37(3):567-575.]
Lohman DJ,Peggie D,Pierce NE,Meier R,2008.Phylogeography and genetic diversity of a widespread Old World butterfly,Lampides boeticus(Lepidoptera:Lycaenidae).Bmc Evolutionary Biology,8(1):301.
Men QL,Xue GX,Mu D,Hu QL,Huang MY,2017.Mitochondrial DNA markers reveal high genetic diversity and strong genetic differentiation in populations of Dendrolimus kikuchii Matsumura(Lepidoptera:Lasiocampidae).PLoS ONE,12(6):e0179706.
Millar CI,Libby WJ,Falk DA,Holsinger KE,1991.Strategies for conserving clinal,ecotypic,and disjunct population diversity in widespread species∥Falk DA,Holsinger KE(eds.).Genetics and Conservation of Rare Plants.England:Oxford University Press.149-170.
Rousset F,1997.Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance.Genetics,145(4):1219-1228.
Rozas J,Sánchez-Delbarrio JC,Messeguer X,Rozas R,2003.DnaSP,DNA polymorphism analyses by the coalescent and other methods.Bioinformatics,19(18):2496-2497.
Excoffier L,Lischer HE,2010.Arlequin suite ver 3.5:a new series of programs to perform population genetics analyses under Linux and Windows.Molecular Ecology Resources,10(3):564-567.
Staden R,1996.The Staden sequence analysis package.Molecular Biotechnology,5(3):233-241.
Sun W,Zhang ZT,Lei CP,Dong H,Qian HT,Cong B,2013.Genetic diversity and genetic differentiation among different geographic populations of Oedaleus infernalis Saussure(Orthoptera:Acridoidea).Journal of Shenyang Agricultural University,44(6):748-753.[孙嵬,张柱亭,类成平,董辉,钱海涛,丛斌,2013.不同地理种群黄胫小车蝗的遗传多样性及遗传分化研究.沈阳农业大学学报,44(6):748-753.]
Tajima F,1989.Statistical method for testing the neutral mutation hypothesis by DNA polymorphism.Genetics,123(3):585-595.
Tamura K,Stecher G,Peterson D,Filipski A,Kumar S,2013.MEGA6:molecular evolutionary genetics analysis verson 6.0.Molecular Biology and Evolution,30(12):2725-2729.
Tang XT,Zheng FS,Lu MX,Du YZ,2014.New ideas about genetic differentiation of Chilo suppressalis(Lepidoptera:Pyralidae)populations in China based on the mtDNA cytochrome b gene.Mitochondrial DNA.Doi:10.3109/19401736.2014.958670.
Wang H,Han LL,Xu ZX,Ye LF,Wang L,Zhao KJ,Wang KQ,2015.Absence of geographic population structure in the soybean pod borer Leguminivora glycinivorella(Tortricidae).Journal of the Lepidopterists’Society,69(3):173-182.
Wang XY,Zhou LH,2016.Genetic diversity and population history among geographic populations of Spodoptera exigua in North China based on mtDNA Cytb gene sequences.Acta Ecologica Sinica,36(8):2337-2347.[王兴亚,周俐宏,2016.基于mtDNA Cytb基因序列的我国北方地区甜菜夜蛾遗传多样性与种群历史分析.生态学报,36(8):2337-2347.]
Zhang SM,Zhao YX,1996.The Geographical Distribution of Agricultural and Forest Insects in China.Beijing:China Agriculture University Press.23-24.[章士美,赵泳祥,1996.中国农林昆虫地理分布.北京:中国农业出版社.23-24.]
Zhang XW,Zhang ZT,Yuan ZG,Zhang QM,Gao BJ,2010.Mitochondrial DNA Cytb gene of insect.Hebei Journal of Forestry and Orchard Research,25(2):173-176.[张学卫,张智婷,原占国,张清敏,高宝嘉,2010.昆虫线粒体DNA Cytb基因研究进展.河北林果研究,25(2):173-176.]
Boivin T,Bouvier JC,Beslay D,Sauphanor B,2004.Variability in diapause propensity within populations of a temperate insect species:interactions between insecticide resistance genes and photoperiodism.Biological Journal of the Linnean Society,83(3):341-351.
Chen QL,Tang XS,Yao WJ,Lu SQ,2009.Bioinformatics analysis of the complete sequences of cytochrome b of Takydromus sylvaticus and modeling the tertiary structure of encoded protein.International Journal of Biological Sciences,5(6):596-602.
Chen YL,Liu JP,1979.The Control of Locust in Xinjiang.Urumqi:Volksverlag Xinjiang.9-10.[陈永林,刘举鹏,1979.新疆蝗虫及其防治.乌鲁木齐:新疆人民出版社.9-10.]
Chen YL,2000.Resurgent control and ecological management of locust.Bulletin of Chinese Academy of Sciences,(5):341-345.[陈永林,2000.蝗虫再猖獗的控制与生态学治理.中国科学院院刊,(5):341-345.]
Dai JX,2005.Cytochrome b gene and molecular systematology research of insects.Sichuan Journal of Zoology,24(2):222-225.[戴金霞,2005.线粒体Cytb基因与昆虫分子系统学研究.四川动物,24(2):222-225.]
Dou J,Zhang RY,Liu M,Qian X,Xiao HW,Roman J,Ji R,2017.Comparison of flight muscle and energy consumption of pre-and post-flight in Calliptamus italicus(Orthoptera:Acrididae).Pratacultural Science,34(8):1721-1726.[窦洁,张若燕,刘敏,钱雪,肖宏伟,Roman Jashenko,季荣,2017.飞行前后意大利蝗飞行肌及能源消耗比较.草业科学,34(8):1721-1726.]
Excoffier L,Lischer HEL,2010.Arlequin suite ver 3.5.A new series of programs to perform population genetics analyses under Linux and Windows.Molecular Ecology Resources,10(3):564-567.
FAO Locust Watch Release,2011.http://www.fao.org/ag/locustsCCA/en/1010/1018/1075/.
Gray MW,1989.Origin and evolution of mitochondrial DNA.Annual Review of Cell Biology,5(5):25.
Guo XZ,1993.Forage resources in the South Mountain area in Urumqi.Xinjiang Animal Husbandry,(4):23-25.[郭选政,1993.乌鲁木齐南山地区的牧草资源.新疆畜牧业,(4):23-25.]
Ha M,Han XH,2014.The current situation of the grassland locusts and the strategies for prevention and treatment in the border area of Tacheng region,Xinjiang.Journal of Agriculture,4(3):7-10.[哈玛尔,韩小虎,2014.新疆塔城地区边境草原蝗虫发生现状及防治对策.农学学报,4(3):7-10.]
Huang H,Zhu EL,2001.The serious outbreak of locusts in Kazakhstan.Word Agriculture,(6):46-47.[黄辉,朱恩林,2001.哈萨克斯坦蝗灾严重发生.世界农业,(6):46-47.]
Kimura M,1980.A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences.Journal of Molecular Evolution,16(2):111-120.
Kokanova EO,2014.The current state of natural foci of swarming locusts(Orthoptera,Acrididae)in Turkmenistan.Entomological Review,94(6):826-828.
Leigh JW,Bryant D,2015.PopART:Full-feature software for haplotype network construction.Methods in Ecology and Evolution,6(9):1110-1116.
Li JB,Ren ZM,2009.Genetic diversity among Schlechtendalia chinensis individuals revealed by Cytb sequences.Journal of Fudan University(Natural Science),48(5):680-686.[李继变,任竹梅,2009.角倍蚜mtDNA Cytb基因遗传多样性分析.复旦学报(自然科学版),48(5):680-686.]
Li PX,Lin J,Mai D,Wu JG,Liu YQ,Zheng JH,2017.Effects of future climate change on potential geographic distribution of Calliptamus italicus(L.)in Xinjiang.Plant Protection,43(3):90-96.[李培先,林峻,麦迪·库尔曼,吴建国,刘永强,郑江华,2017.气候变化对新疆意大利蝗潜在分布的影响.植物保护,43(3):90-96.]
Li R,Han LL,Wang H,Zhao KJ,Mang YL,Zhang HY,Fan D,2015.Analysis of genetic differentiation among different geographic populations of Aphis glycines(Hemiptera:Aphididae)based on mtDNA COII gene sequences.Chinese Journal of Applied Entomology,52(5):1203-1214.[李冉,韩岚岚,王红,赵奎军,莽逸伦,张红玉,樊东,2015.基于线粒体COII基因的大豆蚜不同地理种群遗传分化研究.应用昆虫学报,52(5):1203-1214.]
Liu ZG,Ji T,Shen F,Liang Q,Luo YX,2015.Genetic diversity of geographic populations of Apis cerana cerana estimated by mitochondrial COII gene sequences.Journal of Environmental Entomology,37(3):567-575.[刘振国,吉挺,沈芳,梁勤,罗岳雄,2015.基于线粒体COII基因序列的中华蜜蜂地理种群的遗传多样性研究.环境昆虫学报,37(3):567-575.]
Lohman DJ,Peggie D,Pierce NE,Meier R,2008.Phylogeography and genetic diversity of a widespread Old World butterfly,Lampides boeticus(Lepidoptera:Lycaenidae).Bmc Evolutionary Biology,8(1):301.
Men QL,Xue GX,Mu D,Hu QL,Huang MY,2017.Mitochondrial DNA markers reveal high genetic diversity and strong genetic differentiation in populations of Dendrolimus kikuchii Matsumura(Lepidoptera:Lasiocampidae).PLoS ONE,12(6):e0179706.
Millar CI,Libby WJ,Falk DA,Holsinger KE,1991.Strategies for conserving clinal,ecotypic,and disjunct population diversity in widespread species∥Falk DA,Holsinger KE(eds.).Genetics and Conservation of Rare Plants.England:Oxford University Press.149-170.
Rousset F,1997.Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance.Genetics,145(4):1219-1228.
Rozas J,Sánchez-Delbarrio JC,Messeguer X,Rozas R,2003.DnaSP,DNA polymorphism analyses by the coalescent and other methods.Bioinformatics,19(18):2496-2497.
Excoffier L,Lischer HE,2010.Arlequin suite ver 3.5:a new series of programs to perform population genetics analyses under Linux and Windows.Molecular Ecology Resources,10(3):564-567.
Staden R,1996.The Staden sequence analysis package.Molecular Biotechnology,5(3):233-241.
Sun W,Zhang ZT,Lei CP,Dong H,Qian HT,Cong B,2013.Genetic diversity and genetic differentiation among different geographic populations of Oedaleus infernalis Saussure(Orthoptera:Acridoidea).Journal of Shenyang Agricultural University,44(6):748-753.[孙嵬,张柱亭,类成平,董辉,钱海涛,丛斌,2013.不同地理种群黄胫小车蝗的遗传多样性及遗传分化研究.沈阳农业大学学报,44(6):748-753.]
Tajima F,1989.Statistical method for testing the neutral mutation hypothesis by DNA polymorphism.Genetics,123(3):585-595.
Tamura K,Stecher G,Peterson D,Filipski A,Kumar S,2013.MEGA6:molecular evolutionary genetics analysis verson 6.0.Molecular Biology and Evolution,30(12):2725-2729.
Tang XT,Zheng FS,Lu MX,Du YZ,2014.New ideas about genetic differentiation of Chilo suppressalis(Lepidoptera:Pyralidae)populations in China based on the mtDNA cytochrome b gene.Mitochondrial DNA.Doi:10.3109/19401736.2014.958670.
Wang H,Han LL,Xu ZX,Ye LF,Wang L,Zhao KJ,Wang KQ,2015.Absence of geographic population structure in the soybean pod borer Leguminivora glycinivorella(Tortricidae).Journal of the Lepidopterists’Society,69(3):173-182.
Wang XY,Zhou LH,2016.Genetic diversity and population history among geographic populations of Spodoptera exigua in North China based on mtDNA Cytb gene sequences.Acta Ecologica Sinica,36(8):2337-2347.[王兴亚,周俐宏,2016.基于mtDNA Cytb基因序列的我国北方地区甜菜夜蛾遗传多样性与种群历史分析.生态学报,36(8):2337-2347.]
Zhang SM,Zhao YX,1996.The Geographical Distribution of Agricultural and Forest Insects in China.Beijing:China Agriculture University Press.23-24.[章士美,赵泳祥,1996.中国农林昆虫地理分布.北京:中国农业出版社.23-24.]
Zhang XW,Zhang ZT,Yuan ZG,Zhang QM,Gao BJ,2010.Mitochondrial DNA Cytb gene of insect.Hebei Journal of Forestry and Orchard Research,25(2):173-176.[张学卫,张智婷,原占国,张清敏,高宝嘉,2010.昆虫线粒体DNA Cytb基因研究进展.河北林果研究,25(2):173-176.]