美棘蓟马体内Wolbachia感染的时空分布
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摘要
【目的】明确共生菌Wolbachia在美棘蓟马Echinothrips americanus体内的时空分布动态。【方法】基于Wolbachia的外膜蛋白基因(wsp)序列设计特异引物构建标准质粒,用绝对荧光定量PCR的方法测定美棘蓟马不同发育阶段(卵、若虫、预蛹、蛹、成虫)和雌雄成虫不同组织(头、胸、腹、腹末3节)中Wolbachia的wsp基因拷贝数,分析美棘蓟马共生菌Wolbachia的感染情况。【结果】美棘蓟马体内Wolbachia拷贝数随美棘蓟马发育进程逐渐增大,雌性成虫期Wolbachia拷贝数显著高于卵和若虫期;雌雄成虫不同组织中的拷贝数存在差异,雌性成虫腹部Wolbachia拷贝数显著高于头部、胸部和腹末3节,雄性成虫胸部和腹部Wolbachia拷贝数显著高于头部和腹末3节。美棘蓟马性别和组织与体内Wolbachia拷贝数存在显著的交互作用。【结论】本研究结果表明,美棘蓟马体内的Wolbachia感染状态不仅受宿主发育阶段的影响,而且与性别及分布位置有很大关系,为进一步了解该蓟马的发生、定植和扩散机理提供理论依据。
【Aim 】 To demonstrate spatio-temporal infection dynamics of Wolbachia in the poinsettia thrips,Echinothrips americanus. 【Methods】Specific primers were designed based on the sequence of Wolbachia surface protein gene( wsp) to construct the standard plasmid. The absolute real-time quantitative PCR was used to determine the copy number of wsp in different developmental stages( egg,nymph,prepupa,pupa and adult) and adult tissues( head,thorax,abdomen and terminal three segments of abdomen) of both sexes of the thrips. 【Results】The copy number of Wolbachia increased with the development of E. americanus, and the copy number of Wolbachia in female adult was significantly higher than those in egg and nymphal stages. The copy numbers of Wolbachia in different adult tissues were different. In female adults the copy number of Wolbachia in abdomen was significantly higher than those in head,thorax and the terminal three segments of abdomen,while in male adults the copy numbers in thorax and abdomen were significantly higher than those in head and the terminal three segments of abdomen. Sex and tissue had significant interactions with the copy number of Wolbachia in E. americanus. 【Conclusion】The results indicate that the infection of Wolbachia in E. americanus is affected not only by the developmental stage of host,but also by host sex and tissue. This study provides a theoretical basis for understanding the occurrence,establishment and spreading of this thrips.
【Aim 】 To demonstrate spatio-temporal infection dynamics of Wolbachia in the poinsettia thrips,Echinothrips americanus. 【Methods】Specific primers were designed based on the sequence of Wolbachia surface protein gene( wsp) to construct the standard plasmid. The absolute real-time quantitative PCR was used to determine the copy number of wsp in different developmental stages( egg,nymph,prepupa,pupa and adult) and adult tissues( head,thorax,abdomen and terminal three segments of abdomen) of both sexes of the thrips. 【Results】The copy number of Wolbachia increased with the development of E. americanus, and the copy number of Wolbachia in female adult was significantly higher than those in egg and nymphal stages. The copy numbers of Wolbachia in different adult tissues were different. In female adults the copy number of Wolbachia in abdomen was significantly higher than those in head,thorax and the terminal three segments of abdomen,while in male adults the copy numbers in thorax and abdomen were significantly higher than those in head and the terminal three segments of abdomen. Sex and tissue had significant interactions with the copy number of Wolbachia in E. americanus. 【Conclusion】The results indicate that the infection of Wolbachia in E. americanus is affected not only by the developmental stage of host,but also by host sex and tissue. This study provides a theoretical basis for understanding the occurrence,establishment and spreading of this thrips.
引文
Adachi-Hagimori T,Miura K,Stouthamer R,2008.A new cytogenetic mechanism for bacterial endosymbiont-induced parthenogenesis in Hymenoptera.Proc.R.Soc.Lond.B Biol.Sci.,275(1652):2667-2673.
Arakaki N,Miyoshi T,Noda H,2001.Wolbachia-mediated parthenogenesis in the predatory thrips Fanklintothrips vespiformis(Thysanoptera:Insecta).Proc.R.Soc.Lond.B Biol.Sci.,268(1471):1011-1016.
Baldo L,Lo N,Werren J,2005.Mosaic nature of the Wolbachia surface protein.J.Bacteriol.,187(15):5406-5418.
Baumann L,Baumann P,1994.Growth kinetics of the endosymbiont Buchnera aphidicola in the aphid Schizaphis graminum.Appl.Environ.Microbiol.,60(9):3440-3444.
Bordenstein SR,Marshall ML,Fry AJ,Kim U,Wernegreen JJ,2006.The tripartite associations between bacteriophage,Wolbachia,and arthropods.PLo S Pathog.,2(5):384-393.
Braig H,Zhou W,Dobson S,O'Neill S,1998.Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis.J.Bacteriol.,180(9):2373-2378.
Dobson SL,Bourtzis K,Braig HR,Jones BF,Zhou WG,Rousset F,O'Neill SL,1999.Wolbachia infections are distributed throughout insect somatic and germ line tissues.Insect Biochem.Mol.,29(2):153-160.
Dossi FCA,da Silva EP,C8nsoli FL,2014.Population dynamics and growth rates of endosymbionts during Diaphorina citri(Hemiptera,Liviidae)ontogeny.Microb.Ecol.,68(4):881-889.
Douglas A,Bouvaine S,Russell R,2011.How the insect immune system interacts with an obligate symbiotic bacterium.Proc.R.Soc.Lond.B Biol.Sci.,278(1704):333-338.
Fraune S,Bosch T,2010.Why bacteria matter in animal development and evolution.Bio Essays,32(7):571-580.
Harris HL,Brennan LJ,Keddie BA,Braig HR,2010.Bacterial symbionts in insects:balancing life and death.Symbiosis,51(1):37-53.
Hilgenboecker K,Hammerstein P,Schlattmann P,Telschow A,Werren J,2008.How many species are infected with Wolbachia?-a statistical analysis of current data.FEMS Microbiol.Lett.,281(2):215-220.
Hoffmann M,Coy M,Gibbard HK,Pelz-Stelinski K,2014.Wolbachia infection density in populations of the Asian citrus psyllid(Hemiptera:Liviidae).Environ.Entomol.,43(5):1215-1222.
Kaas JP,2001.Scouting for thrips-the development of a time saving sampling program for Echinothrips.Proc.Exp.Appl.Entomol.,12:85-90.
Kageyama D,Wang CH,Hatakeyama M,2017.Wolbachia infections of the butterfly Eurema mandarina interfere with embryonic development of the sawfly Athalia rosae.J.Invertebr.Pathol.,150(Suppl.C):76-81.
Kozek W,Rao RU,2007.The discovery of Wolbachia in arthropods and nematodes-a historical perspective.In:Hoerauf A,Rao RU eds.Wolbachia:A Bug’s Life in Another Bug.Issues in Infectious Diseases,Vol.5.Basel,Karger.1-14.
Kumm S,Moritz G,2008.First detection of Wolbachia in arrhenotokous populations of thrips species(Thysanoptera:Thripidae and Phlaeothripidae)and its role in reproduction.Environ.Entomol.,37(6):1422-1428.
Kusmintarsih ES,2011.Ultrastucture of Wolbachia are found in somatic and reproductive tissue of Drosophila simulans and D.melanogaster.Microbiol.Indones.,5(1):33-38.
Kyei-Poku GK,Giladi M,Coghlin P,Mokady O,Zchori-Fein E,Floate KD,2006.Wolbachia in wasps parasitic on filth flies with emphasis on Spalangia cameroni.Entomol.Exp.Appl.,121(2):123-135.
Landmann F,Slatko B,Foster JM,Sullivan W,2010.Asymmetric Wolbachia segregation during early Brugia malayi embryogenesis determines its distribution in adult host tissues.PLo S Negl.Trop.Dis.,4(7):e758.
Li HF,Li L,Zhang LJ,Xu YL,Li WF,2010.Standard curve generation of Pep T1 gene for absolute quantification using real-time PCR.J.Shanxi Agric.Univ.(Nat.Sci.Ed.),30(4):332-335.[李慧锋,李丽,张丽娟,徐玉良,李武峰,2010.Pep T1基因绝对荧光定量PCR标准曲线的建立.山西农业大学学报(自然科学版),30(4):332-335]
Li XW,Zhang XC,Jiang HX,Feng JN,2012.Comparisons of developmental and reproductive biology between parthenogenetic and sexual Echinothrips americanus(Thysanoptera:Thripidae).Environ.Entomol.,41(3):706-713.
Mc Graw EA,Merritt DJ,Droller JN,O'Neill SL,2002.Wolbachia density and virulence attenuation after transfer into a novel host.Proc.Natl.Acad.Sci.USA,99(5):2918-2923.
Mirab-Balou M,Lu H,Chen XX,2010.First record of Echinothrips americanus Morgan(Thysanoptera,Thripidae)in mainland China.Acta Zootaxon.Sin.,35(3):674-679.
Ndii MZ,Hickson RI,Allingham D,Mercer GN,2015.Modelling the transmission dynamics of dengue in the presence of Wolbachia.Math.Biosci.,262:157-166.
Oetting RD,Beshear RJ,1993.Biology of the greenhouse pest Echinothrips americanus Morgan(Thysanoptera:Thripidae).Int.J.Pure Appl.Zool.,4:307-315.
Opit GP,Peterson B,Gillespie DR,Costello RA,1997.The life cycle and management of Echinothrips americanus.J.Entomol.Soc.Brit.Columba,94:3-6.
Ratzka C,Gross R,Feldhaar H,2012.Endosymbiont tolerance and control within insect hosts.Insects,3(4):553-572.
Rigaud T,Rousset F,1996.What generates the diversity of Wolbachiaarthropod interactions?Biodivers.Conserv.,5(8):999-1013.
Roy V,Girondot M,Harry M,2015.The distribution of Wolbachia in Cubitermes(Termitidae,Termitinae)castes and colonies:a modelling approach.PLo S ONE,10(2):e0116070.
Sacchi L,Corona S,Grigolo A,Laudani U,Selmi M,Bigliard E,1996.The fate of the endocytobionts of Blattella germanica(Blattaria:Blattellidae)and Periplaneta americana(Blattaria:Blattidae)during embryo development.Ital.J.Zool.,63(1):1-11.
Shen JF,Ming QL,Cheng C,Liu CM,Feng ZJ,2016.The temporal and spatial distribution status of Wolbachia infection density in the confused flour beetle Tribolium confusum.J.Environ.Entomol.,38(2):378-383.[沈加飞,明庆磊,程超,刘缠民,冯照军,2016.杂拟谷盗体内Wolbachia感染密度的时间和空间分布状况.环境昆虫学报,38(2):378-383]
Stannard LJ,1968.The thrips,or Thysanoptera,of Illinois.Bull.Illinois Nat.Hist.Survey,29:307-308.
Timmermans M,Ellers J,2009.Wolbachia endosymbiont is essential for egg hatching in a parthenogenetic arthropod.Evol.Ecol.,23(6):931-942.
Varga L,Fedor PJ,2008.First interception of the greenhouse pest Echinothrips americanus Morgan,1913(Thysanoptera:Thripidae)in Slovak Republic.Plant Prot.Sci.,44(4):155-158.
Veneti Z,Clark ME,Karr TL,Savakis C,Bourtzis K,2004.Heads or tails:host-parasite interactions in the Drosophila-Wolbachia system.Appl.Environ.Microb.,70(9):5366-5372.
Wang CY,2011.Genetic mechanism and evolutionary significance of the origin of parthenogenetic insects.Zool.Res.,32(6):689-695.[王成业,2011.昆虫孤雌生殖起源的遗传机制和进化意义.动物学研究,32(6):689-695]
Wang Z,Qiao GX,2011.Current research trends on the endosymbion Wolbachia in insects.Chin.J.Appl.Entomol.,48(6):1823-1834.[王哲,乔格侠,2011.Wolbachia与昆虫寄主关系研究进展.应用昆虫学报,48(6):1823-1834]
Werren JH,1997.Biology of Wolbachia.Annu.Rev.Entomol.,42:587-609.
Werren JH,Windsor D,Guo LR,1995.Distriution of Wolbachia among neotropical arthropods.Proc.R.Soc.Lond.B Biol.Sci.,262(1364):197-204.
Wolschin F,Holldobler B,Gross R,Zientz E,2004.Replication of the endosymbiotic bacterium Blochmannia floridanus is correlated with the developmental and reproductive stages of its ant host.Appl.Environ.Microbiol.,70(7):4096-4102.
Zhang H,Zhang KJ,Hong XY,2010.Infection density of Wolbachia in different tissues of macropters and brachypters adults of Nilaparvata lugens(Stl).J.Nanjing Agric.Univ.,33(5):35-39.[张慧,张开军,洪晓月,2010.褐飞虱长短翅型成虫不同组织内Wolbachia的密度.南京农业大学学报,33(5):35-39]
Zhang XC,2010.New Record of Echinothrips americanus Morgan(Thysanoptera:Thripidae)in China Mainland and Its Biology.MSc Thesis,Northwest A&F University,Yangling,Shaanxi.[张晓晨,2010.中国大陆新记录种美棘蓟马Echinothrips americanus Morgan及其生物学研究.陕西杨凌:西北农林科技大学硕士学位论文]
Zheng Y,Wang JL,Liu C,Wang CP,Walker T,Wang YF,2011.Differentially expressed profiles in the larval testes of Wolbachia infected and uninfected Drosophila.BMC Genomics,12:595.
Arakaki N,Miyoshi T,Noda H,2001.Wolbachia-mediated parthenogenesis in the predatory thrips Fanklintothrips vespiformis(Thysanoptera:Insecta).Proc.R.Soc.Lond.B Biol.Sci.,268(1471):1011-1016.
Baldo L,Lo N,Werren J,2005.Mosaic nature of the Wolbachia surface protein.J.Bacteriol.,187(15):5406-5418.
Baumann L,Baumann P,1994.Growth kinetics of the endosymbiont Buchnera aphidicola in the aphid Schizaphis graminum.Appl.Environ.Microbiol.,60(9):3440-3444.
Bordenstein SR,Marshall ML,Fry AJ,Kim U,Wernegreen JJ,2006.The tripartite associations between bacteriophage,Wolbachia,and arthropods.PLo S Pathog.,2(5):384-393.
Braig H,Zhou W,Dobson S,O'Neill S,1998.Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis.J.Bacteriol.,180(9):2373-2378.
Dobson SL,Bourtzis K,Braig HR,Jones BF,Zhou WG,Rousset F,O'Neill SL,1999.Wolbachia infections are distributed throughout insect somatic and germ line tissues.Insect Biochem.Mol.,29(2):153-160.
Dossi FCA,da Silva EP,C8nsoli FL,2014.Population dynamics and growth rates of endosymbionts during Diaphorina citri(Hemiptera,Liviidae)ontogeny.Microb.Ecol.,68(4):881-889.
Douglas A,Bouvaine S,Russell R,2011.How the insect immune system interacts with an obligate symbiotic bacterium.Proc.R.Soc.Lond.B Biol.Sci.,278(1704):333-338.
Fraune S,Bosch T,2010.Why bacteria matter in animal development and evolution.Bio Essays,32(7):571-580.
Harris HL,Brennan LJ,Keddie BA,Braig HR,2010.Bacterial symbionts in insects:balancing life and death.Symbiosis,51(1):37-53.
Hilgenboecker K,Hammerstein P,Schlattmann P,Telschow A,Werren J,2008.How many species are infected with Wolbachia?-a statistical analysis of current data.FEMS Microbiol.Lett.,281(2):215-220.
Hoffmann M,Coy M,Gibbard HK,Pelz-Stelinski K,2014.Wolbachia infection density in populations of the Asian citrus psyllid(Hemiptera:Liviidae).Environ.Entomol.,43(5):1215-1222.
Kaas JP,2001.Scouting for thrips-the development of a time saving sampling program for Echinothrips.Proc.Exp.Appl.Entomol.,12:85-90.
Kageyama D,Wang CH,Hatakeyama M,2017.Wolbachia infections of the butterfly Eurema mandarina interfere with embryonic development of the sawfly Athalia rosae.J.Invertebr.Pathol.,150(Suppl.C):76-81.
Kozek W,Rao RU,2007.The discovery of Wolbachia in arthropods and nematodes-a historical perspective.In:Hoerauf A,Rao RU eds.Wolbachia:A Bug’s Life in Another Bug.Issues in Infectious Diseases,Vol.5.Basel,Karger.1-14.
Kumm S,Moritz G,2008.First detection of Wolbachia in arrhenotokous populations of thrips species(Thysanoptera:Thripidae and Phlaeothripidae)and its role in reproduction.Environ.Entomol.,37(6):1422-1428.
Kusmintarsih ES,2011.Ultrastucture of Wolbachia are found in somatic and reproductive tissue of Drosophila simulans and D.melanogaster.Microbiol.Indones.,5(1):33-38.
Kyei-Poku GK,Giladi M,Coghlin P,Mokady O,Zchori-Fein E,Floate KD,2006.Wolbachia in wasps parasitic on filth flies with emphasis on Spalangia cameroni.Entomol.Exp.Appl.,121(2):123-135.
Landmann F,Slatko B,Foster JM,Sullivan W,2010.Asymmetric Wolbachia segregation during early Brugia malayi embryogenesis determines its distribution in adult host tissues.PLo S Negl.Trop.Dis.,4(7):e758.
Li HF,Li L,Zhang LJ,Xu YL,Li WF,2010.Standard curve generation of Pep T1 gene for absolute quantification using real-time PCR.J.Shanxi Agric.Univ.(Nat.Sci.Ed.),30(4):332-335.[李慧锋,李丽,张丽娟,徐玉良,李武峰,2010.Pep T1基因绝对荧光定量PCR标准曲线的建立.山西农业大学学报(自然科学版),30(4):332-335]
Li XW,Zhang XC,Jiang HX,Feng JN,2012.Comparisons of developmental and reproductive biology between parthenogenetic and sexual Echinothrips americanus(Thysanoptera:Thripidae).Environ.Entomol.,41(3):706-713.
Mc Graw EA,Merritt DJ,Droller JN,O'Neill SL,2002.Wolbachia density and virulence attenuation after transfer into a novel host.Proc.Natl.Acad.Sci.USA,99(5):2918-2923.
Mirab-Balou M,Lu H,Chen XX,2010.First record of Echinothrips americanus Morgan(Thysanoptera,Thripidae)in mainland China.Acta Zootaxon.Sin.,35(3):674-679.
Ndii MZ,Hickson RI,Allingham D,Mercer GN,2015.Modelling the transmission dynamics of dengue in the presence of Wolbachia.Math.Biosci.,262:157-166.
Oetting RD,Beshear RJ,1993.Biology of the greenhouse pest Echinothrips americanus Morgan(Thysanoptera:Thripidae).Int.J.Pure Appl.Zool.,4:307-315.
Opit GP,Peterson B,Gillespie DR,Costello RA,1997.The life cycle and management of Echinothrips americanus.J.Entomol.Soc.Brit.Columba,94:3-6.
Ratzka C,Gross R,Feldhaar H,2012.Endosymbiont tolerance and control within insect hosts.Insects,3(4):553-572.
Rigaud T,Rousset F,1996.What generates the diversity of Wolbachiaarthropod interactions?Biodivers.Conserv.,5(8):999-1013.
Roy V,Girondot M,Harry M,2015.The distribution of Wolbachia in Cubitermes(Termitidae,Termitinae)castes and colonies:a modelling approach.PLo S ONE,10(2):e0116070.
Sacchi L,Corona S,Grigolo A,Laudani U,Selmi M,Bigliard E,1996.The fate of the endocytobionts of Blattella germanica(Blattaria:Blattellidae)and Periplaneta americana(Blattaria:Blattidae)during embryo development.Ital.J.Zool.,63(1):1-11.
Shen JF,Ming QL,Cheng C,Liu CM,Feng ZJ,2016.The temporal and spatial distribution status of Wolbachia infection density in the confused flour beetle Tribolium confusum.J.Environ.Entomol.,38(2):378-383.[沈加飞,明庆磊,程超,刘缠民,冯照军,2016.杂拟谷盗体内Wolbachia感染密度的时间和空间分布状况.环境昆虫学报,38(2):378-383]
Stannard LJ,1968.The thrips,or Thysanoptera,of Illinois.Bull.Illinois Nat.Hist.Survey,29:307-308.
Timmermans M,Ellers J,2009.Wolbachia endosymbiont is essential for egg hatching in a parthenogenetic arthropod.Evol.Ecol.,23(6):931-942.
Varga L,Fedor PJ,2008.First interception of the greenhouse pest Echinothrips americanus Morgan,1913(Thysanoptera:Thripidae)in Slovak Republic.Plant Prot.Sci.,44(4):155-158.
Veneti Z,Clark ME,Karr TL,Savakis C,Bourtzis K,2004.Heads or tails:host-parasite interactions in the Drosophila-Wolbachia system.Appl.Environ.Microb.,70(9):5366-5372.
Wang CY,2011.Genetic mechanism and evolutionary significance of the origin of parthenogenetic insects.Zool.Res.,32(6):689-695.[王成业,2011.昆虫孤雌生殖起源的遗传机制和进化意义.动物学研究,32(6):689-695]
Wang Z,Qiao GX,2011.Current research trends on the endosymbion Wolbachia in insects.Chin.J.Appl.Entomol.,48(6):1823-1834.[王哲,乔格侠,2011.Wolbachia与昆虫寄主关系研究进展.应用昆虫学报,48(6):1823-1834]
Werren JH,1997.Biology of Wolbachia.Annu.Rev.Entomol.,42:587-609.
Werren JH,Windsor D,Guo LR,1995.Distriution of Wolbachia among neotropical arthropods.Proc.R.Soc.Lond.B Biol.Sci.,262(1364):197-204.
Wolschin F,Holldobler B,Gross R,Zientz E,2004.Replication of the endosymbiotic bacterium Blochmannia floridanus is correlated with the developmental and reproductive stages of its ant host.Appl.Environ.Microbiol.,70(7):4096-4102.
Zhang H,Zhang KJ,Hong XY,2010.Infection density of Wolbachia in different tissues of macropters and brachypters adults of Nilaparvata lugens(Stl).J.Nanjing Agric.Univ.,33(5):35-39.[张慧,张开军,洪晓月,2010.褐飞虱长短翅型成虫不同组织内Wolbachia的密度.南京农业大学学报,33(5):35-39]
Zhang XC,2010.New Record of Echinothrips americanus Morgan(Thysanoptera:Thripidae)in China Mainland and Its Biology.MSc Thesis,Northwest A&F University,Yangling,Shaanxi.[张晓晨,2010.中国大陆新记录种美棘蓟马Echinothrips americanus Morgan及其生物学研究.陕西杨凌:西北农林科技大学硕士学位论文]
Zheng Y,Wang JL,Liu C,Wang CP,Walker T,Wang YF,2011.Differentially expressed profiles in the larval testes of Wolbachia infected and uninfected Drosophila.BMC Genomics,12:595.