Biological Invasions

, Volume 13, Issue 12, pp 2869–2887 | Cite as

Genetic analysis of emerald ash borer (Agrilus planipennis Fairmaire) populations in Asia and North America

  • Alicia M. Bray
  • Leah S. Bauer
  • Therese M. Poland
  • Robert A. Haack
  • Anthony I. Cognato
  • James J. SmithEmail author
Original Paper


Emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is an invasive pest of North American ash (Fraxinus spp.) trees first discovered outside of its native range of northeastern Asia in 2002. EAB spread from its initial zone of discovery in the Detroit, Michigan and Windsor, Ontario metropolitan areas, in large part, from inadvertent human-assisted movement of infested ash materials. EAB infestations are now known in 15 US states and two Canadian provinces. The primary goal of this study was to use molecular markers to characterize the population genetic structure of EAB in its native and introduced range. This information may provide valuable insights on the geographic origin, potential host range, invasion potential, and additional biological control agents for ongoing management efforts of this destructive wood-boring beetle. EAB were collected from 17 localities in its native Asian range and from 7 localities in North America, and population structure analyzed using mtDNA gene sequences, AFLP fingerprints, and alleles at 2 microsatellite loci. Analysis of mtDNA cytochrome oxidase subunit I gene (COI; 439 bp) sequences revealed all North American individuals carry a common mtDNA haplotype also found in China and South Korea. Additional mtDNA haplotypes observed in China and South Korea differed from the common haplotype by 1–2 nucleotide substitutions and a single individual from Japan differed by 21 nucleotide changes (4.8%). Analysis using AFLP fingerprints (108 loci) indicated Asian populations were more highly variable, yet had less overall population structure, than the North American populations. North American populations appear most closely related to populations in our sample from the Chinese provinces of Hebei and Tianjin City. Further, population assignment tests assigned 88% of the individual beetles from North America to either Hebei or Tianjin City.


Agrilus planipennis Population genetics mtDNA AFLP Microsatellites Invasive species 



Many thanks to the network of collaborators who helped us obtain the insects used in this study, including countless members in North America from USDA Forest Service Northern Research Station, Michigan State University, Illinois, Indiana, and Michigan Departments of Agriculture, and the Canadian Food Inspection Agency. We would like to specifically thank collaborators in Asia and the US who made sampling possible, in China: Dr. Houping Liu (Michigan State University, Dr. Tonghai Zhao and Dr. Ruitong Gao (Chinese Academy of Forestry), and Mr. Roger Fuester (USDA ARS BIIR); in South Korea: Dr. David Williams (USDA APHIS CPHST), Dr. Choo Ho Yul (Gyeongsang National University) and Dr. Lee DongWoon (Kyungpook National University); in Japan—Dr. Paul Schaefer (USDA-ARS-BIIR), Dr. Naoto Kamata (University of Tokyo), Mr. Minemitsu Kaneko (Japan Wildlife Research Center), and Mr. Takaharu Hattori. We also thank Deborah Miller (USDA Forest Service, Northern Research Station, E. Lansing, MI) for organizing and managing the long-term storage of EAB collections from China and North America; Dr. Jennifer Cory (Simon Fraser University) for designing and developing the microsatellite primers used in the study, Dr. Deb McCullough (Michigan State University) for supporting the early microsatellite development work, Dr. Rob Ahern for his help with AFLP data analysis and interpretation, Mike Grillo for his work during the earliest phases of the EAB project, Dr. Rodrigo Mercador and Dr. Yigen Chen for comments on an earlier version of the manuscript, and Marlene Cameron for assistance with the preparation of Figure 1. Research funding for the project was provided by the USDA Forest Service (03-JV-11231300096), Michigan State University (MSU) Project GREEEN (Generating Research and Extension to meet Economic and Environmental Needs), the MSU Office for International Studies and Programs, the MSU Department of Entomology, and the MSU Graduate School.


  1. Abdelkrim J, Pascal M, Calmet C, Samadi S (2005) Importance of assessing population genetic structure before eradication of invasive species: examples from insular Norway rat populations. Conserv Biol 19:1509–1518CrossRefGoogle Scholar
  2. Ahern RG, Hawthorne DJ, Raupp MJ (2009) Phylogeography of a specialist insect, Adelges cooleyi: historical and contemporary processes shape the distribution of population genetic variation. Molec Ecol 18:343–356CrossRefGoogle Scholar
  3. Akiyama K, Ohmomo S (1997) A checklist of the Japanese Buprestidae. Gekkan-Mushi (1) pp 1–67Google Scholar
  4. Allendorf FW, Lundquist LL (2003) Introduction: population biology, evolution, and control of Invasive species. Conserv Biol 17:24–30CrossRefGoogle Scholar
  5. Anulewicz AC, McCullough DG, Cappaert DL, Poland TM (2008) Host range of the emerald ash borer (Agrilus planipennis Fairmaire) (Coleoptera: Buprestidae) in North America: results of multiple-choice field experiments. Environ Entomol 37:230–241PubMedCrossRefGoogle Scholar
  6. Baranchikov Y, Mozolevskaya E, Yurchenko G, Kenis M (2008) Occurrence of the emerald ash borer, Agrilus planipennis in Russia and its potential impact on European forestry. EPPO Bull 38:233–238CrossRefGoogle Scholar
  7. Bauer LS, Liu H, Haack RA, Petrice TR, Miller DL (2004) Natural enemies of emerald ash borer in southeastern Michingan. In: Proceedings of the 2003 Emerald Ash Borer Research and Technology Development Meeting, September 30—October 01, 2003, Port Huron, Michigan. USDA Forest Service, FHTET Publ, 2004–2002. p 33Google Scholar
  8. Bauer LS, Liu H-P, Haack RA, Gao R, Zhao T, Miller DL, Petrice TR (2005) Emerald ash borer natural enemy surveys in Michigan and China. In: Mastro V, Reardon R (Compilers), Proceedings of Emerald Ash Borer Research and Technology Development Meeting, Romulus, MI USDA FS FHTET-2004-15, pp 71–72
  9. Bauer LS, Liu H-P, Miller DL, Gould JR (2008) Developing a classical biological control program for Agrilus planipennis (Coleoptera: Buprestidae), an invasive ash pest in North America. Newsletter Mich Entomol Soc 53:38–39
  10. Bauer LS, Gould J, Duan J (In press) Emerald ash borer biological control. In: Proceedings, 20th US Department of Agriculture interagency research forum on gypsy moth and other invasive species. USDA FS NRS, Gen Tech RepGoogle Scholar
  11. Benavides P, Vega FE, Romero-Severson J, Bustillo AE, Stuart JJ (2005) Biodiversity and biogeography of an important inbred pest of coffee, coffee berry borer (Coleoptera: Curculionidae: Scolytinae). Ann Entomol Soc Am 98:359–366CrossRefGoogle Scholar
  12. Bonizzoni M, Zheng L, Guglielmino CR, Haymer DS, Gasperi G, Gomulski LM, Malacrida AR (2001) Microsatellite analysis of medfly bioinfestations in California. Molec Ecol 10:2515–2524CrossRefGoogle Scholar
  13. Bray AM (2009) AFLP, mtDNA, and microsatellite analysis of emerald ash borer population structure from Asia and North America. Dissertation, Michigan State UniversityGoogle Scholar
  14. Bray AM, Bauer LS, Haack RA, Poland T, Smith JJ (2008) Emerald ash borer genetics: an update. In: Proceedings of the 2007 Emerald Ash Borer Research and Technology Development Meeting, October 23–24, 2007, Pittsburgh, Pennsylvania. USDA Forest Service, FHTET Publ. 2008–2007: p 11Google Scholar
  15. Cappaert D, McCullough DG, Poland TM, Siegert NW (2005) Emerald ash borer in North America: a research and regulatory challenge. Am Entomol 51:152–165Google Scholar
  16. Chinese Academy of Science, Institute of Zoology (1986) Agrilus marcopoli Obenberger. In: Editorial committee (eds.) Agriculture Insects of China (part I). China Agriculture Press, Beijing, China, p 445Google Scholar
  17. Clement M, Posada D, Crandall K (2000) TCS: a computer program to estimate gene genealogies. Molec Ecol 9:1657–1660CrossRefGoogle Scholar
  18. Cognato AI, Sun J-H, Anducho-Reyes MA, Owen DR (2005) Genetic variation and origin of red turpentine beetle (Dendroctonus valens LeConte) introduced to the People’s Republic of China. Agric For Entomol 7:87–94CrossRefGoogle Scholar
  19. Conner JK, Hartl DL (2004) A primer of ecological genetics. Sinauer Associates, Inc. Sutherland, MassachusettsGoogle Scholar
  20. Conord C, Lemperiere G, Taberlet P, Despres L (2006) Genetic structure of the forest pest Hylobius abietis on conifer plantations at different spatial scales in Europe. Heredity 97:46–55PubMedCrossRefGoogle Scholar
  21. Cooperative Emerald Ash Borer Project (2010); Accessed June 2010
  22. Dieringer D, Schlotterer C (2003) Microsatellite Analyser (MSA): a platform independent analysis tool for large microsatellite data sets. Molec Ecol Notes 3:167–169CrossRefGoogle Scholar
  23. Duchesne P, Bernatchez L (2002) AFLPOP: a computer program for simulated and real population allocation based on AFLP data. Molec Ecol Notes 2:380–383CrossRefGoogle Scholar
  24. Elderkin CL, Perkins EJ, Leberg PL, Klerks PL, Lance RF (2004) Amplified fragment length polymorphism (AFLP) analysis of the genetic structure of the zebra mussel, Dreissena polymorpha, in the Mississippi River. Freshw Biol 49:1487–1494CrossRefGoogle Scholar
  25. Elkinton JS, Liebhold AM (1990) Population dynamics of Gypsy moth in North America. Ann Rev Entomol 35:571–596CrossRefGoogle Scholar
  26. Excoffier L, Laval G, Schneider S (2005) Arlequin, Version 3.0: An integrated software package for population genetic data analysis. Evol Bioinforma Online 1:47–50Google Scholar
  27. Federal Register (2007) The proposed release of three parasitoids for the biological control of the emerald ash borer (Agrilus planipennis) in the continental United States: environmental assessment. Federal Register 72:28947–28948, Docket No. APHIS-2007-0060 Google Scholar
  28. Felsenstein J (2005) PHYLIP (Phylogeny Inference Package) version 3.6. Distributed by the author. Department of Genome Sciences, University of Washington, SeattleGoogle Scholar
  29. Fu D-Y, Hu S-J, Ye H, Haack RA, Zhou P-Y (2010) Pine wilt disease in Yunnan, China: evidence of non-local pine sawyer Monochamus alternatus (Coleoptera: Cerambycidae) populations revealed by mitochondrial DNA. Insect Science 17:439–447Google Scholar
  30. Genton BJ, Shykoff JA, Giraud T (2005) High genetic diversity in French invasive populations of common ragweed, Ambrosia artemisiifolia, as a result of multiple sources of introduction. Molec Ecol 14:4275–4285CrossRefGoogle Scholar
  31. Grapputo A, Boman S, Lindstrom L, Lyytinen A, Mappes J (2005) The voyage of an invasive species across continents: genetic diversity of North American and European Colorado potato beetle populations. Molec Ecol 14:4207–4219CrossRefGoogle Scholar
  32. Haack RA, Jendek E, Liu H, Marchant K, Petrice T, Poland T, Ye H (2002) The emerald ash borer: a new exotic pest in North America. Newsletter Mich Entomol Soc 47(3–4):1–5Google Scholar
  33. Hou T-Q (1993) Agrilus marcopoli Obenberger. In: Editorial committee (eds.) Fauna of Shandong Forest Insect. China Forestry publishing House. Beijing, China. p 237, (in Chinese) Google Scholar
  34. Jendek E (1994) Studies in the East Palaearctic species of the genus Agrilus Dahl, 1823 (Coleoptera: Buprestidae) Part I. Entomol Problems 25:9–25Google Scholar
  35. Kambhampati S, Rai KS (1991) Mitochondrial DNA variation within and among populations of the mosquito Aedes albopictus. Genome 34:288–292PubMedCrossRefGoogle Scholar
  36. Kim KS, Cano-Rios P, Sappington TW (2006) Using genetic markers and population assignment techniques to infer origin of boll weevils (Coleoptera: Curculionidae) unexpectedly captured near an eradication zone in Mexico. Environ Entomol 35:813–826CrossRefGoogle Scholar
  37. Lee CE (2002) Evolutionary genetics of invasive species. Trends Ecol Evol 17:386–391CrossRefGoogle Scholar
  38. Liu H, Bauer LS, Gap R, Zhao T, Petrice TR, Haack RA (2003) Exploratory survey for the emerald ash borer, Agrilus planipennis (Coleoptera: Buprestidae) and its natural enemies in China. Grt Lakes Entomol 36:191–204Google Scholar
  39. Liu H-P, Bauer LS, Miller DL, Zhao TH, Gao R-T, Song L, Luan Q, Jin R, Gao C (2007) Seasonal abundance of Agrilus planipennis (Coleoptera: Buprestidae) and its natural enemies Oobius agrili (Hymenoptera: Encyrtidae) and Tetrastichus planipennisi (Hymenoptera: Eulophidae) in China. Biol Control 42:61–71CrossRefGoogle Scholar
  40. Lynch M, Milligan BG (1994) Analysis of population genetic structure with RAPD markers. Molec Ecol 3:91–99CrossRefGoogle Scholar
  41. McCullough DG, Schneeberger NF, Katovich SA (2008) Pest alert: emerald ash borer. USDA Foret Service, Northeastern Area State and Private Forestry, NA-PR-02-04. Accessed Feb 2010
  42. Mueller UG, Wolfenbarger LL (1999) AFLP genotyping and fingerprinting. Trends Ecol Evol 14:389–394PubMedCrossRefGoogle Scholar
  43. Paupy C, Orsoni A, Mousson L, Huber K (2004) Comparisons of amplified fragment length polymorphism (AFLP), microsatellite, and isoenzyme markers: population genetics of Aedes aegypti (Diptera: Culicidae) from Phnom Penh (Cambodia). J Med Entomol 41:664–671PubMedCrossRefGoogle Scholar
  44. Peakall R, Smouse PE (2006) GenAlEx 6: genetic analysis in Excel. Population genetic software for teaching and research. Molec Ecol Notes 6:288–295CrossRefGoogle Scholar
  45. Poland TM, McCullough DG (2006) Emerald ash borer: invasion of the urban forest and the threat to North America’s ash resource. J Forestry April-May:118–124Google Scholar
  46. Reineke A, Karlovsky P, Zebitz CPW (1999) Amplified fragment length polymorphism analysis of different grographic populations of the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae). Bull Entomol Research 89:79–88Google Scholar
  47. Roderick GK (1996) Geographic structure of insect populations: gene flow, phylogeography, and their uses. Ann Rev Entomol 41:325–352CrossRefGoogle Scholar
  48. Ross KG, Krieger JB, Keller L, Shoemaker DD (2007) Genetic variation and structure in native populations of the fire ant Solenopsis invicta: evolutionary and demographic implications. Biol J Linn Soc 92:541–560CrossRefGoogle Scholar
  49. Sakai AK, Allendorf FW, Holt JS, Lodge DM, Molofsky J, With KA, Baughman S, Cabin RJ, Cohen JE, Ellstrand NC, McCauley DE, O’Neil P, Parker IM, Thompson JN, Weller SG (2001) The population biology of invasive species. Ann Rev Ecol Syst 32:305–322CrossRefGoogle Scholar
  50. Salvato P, Battisti A, Concato S, Masutti L, Patarnello T, Zane L (2002) Genetic differentiation in the winter pine processionary moth (Thaumetopoea pityocampa - wilkinsoni complex), inferred by AFLP and mitochondrial DNA markers. Molec Ecol 11:2435–2444CrossRefGoogle Scholar
  51. Scheffer SJ, Grissell EE (2003) Tracing the geographical origin of Megastigmus transvaalensis (Hymenoptera: Torymidae): an African wasp feeding on a South American plant in North America. Molec Ecol 12:415–421CrossRefGoogle Scholar
  52. Shoemaker DD, Ahrens ME, Ross KG (2006) Molecular phylogeny of fire ants of the Solenopsis saevissima species-group based on mtDNA sequences. Molec Phylogenetics Evol 38:200–215CrossRefGoogle Scholar
  53. Siegert NW, McCullough DG, Liebhold AM, Telewski FW (2007) Dendochronological reconstruction of the establishment and spread of emerald ash borer. In: Forest Health Technology Enterprise Team: Emerald ash borer research and technology development meetingGoogle Scholar
  54. Siegert NW, McCullough DG, Williams DW, Fraser I, Poland TM, Pierce SJ (2010) Dispersal of Agrilus planipennis (Coleoptera: Buprestidae) from discrete epicenters in two outlier sites. Environ Entomol 39:253–265PubMedCrossRefGoogle Scholar
  55. Simon C, Frati F, Beckenbach A, Crespi B, Liu H, Flook P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Ann Entomol Soc Am 87:651–701Google Scholar
  56. Szalanski AL, Owens CB (2003) Genetic variation of the southern corn rootworm, (Coleoptera: Chrysomelidae). Fla Entomol 86:329–333CrossRefGoogle Scholar
  57. Taylor RAJ, Bauer LS, Poland TM, Windell K (2010) Flight performance of Agrilus planipennis (Coleoptera: Buprestidae) on a flight mill and in free flight. J Insect Behavior 23:128–148CrossRefGoogle Scholar
  58. Tsutsui ND, Suarez AV, Holway DA, Case TJ (2001) Relationships among native and introduced populations of the Argentine ant (Linepithema humile) and the source of introduced populations. Molec Ecol 10:2151–2161CrossRefGoogle Scholar
  59. USDA-APHIS (2010) Emerald ash borer quarantine map. Accessed Feb 2010
  60. Vekemans X, Beauwens T, Lemaire M, Roldan-Ruiz I (2002) Data from amplified fragment length polymorphism (AFLP) markers show indication of size homoplasy and of a relationship between degree of homoplasy and fragment size. Molec Ecol 11:139–151CrossRefGoogle Scholar
  61. Vos P, Bleeker M, Reijans M, VanDelee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP-A new technique for DNA-Fingerprinting. Nucleic Acids Res 23:4407–4414PubMedCrossRefGoogle Scholar
  62. Wei X, Reardon D, Wu Y, Sun JH (2004) Emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera; Buprestidae), in China: a review and distribution survey. Acta Entomol Sinica 47:679–685Google Scholar
  63. Wright S (1951) The genetic structure of populations. Ann Eugen 15:323–354CrossRefGoogle Scholar
  64. Xu G-T (2003) Agrilus marcopoli Obenberger. Atlas of ornamental pests and Diseases. China Agriculture Press, Beijing, pp 321–322Google Scholar
  65. Yu C (1992) Agrilus marcopoli Obenberger. Pages 400–401 in G. Xiao [ed.], Forest Insects of China, 2nd edn. China Forestry Publishing House, Beijing, ChinaGoogle Scholar
  66. Zhang Y, Huang D, Zhao T, Liu H, Bauer LS (2005) Two new species of egg parasitoids (Hymenoptera: Encyrtidae) of wood-boring beetle pests from China. Phytoparasitica 33:253–260CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Alicia M. Bray
    • 1
    • 4
  • Leah S. Bauer
    • 2
  • Therese M. Poland
    • 2
  • Robert A. Haack
    • 2
  • Anthony I. Cognato
    • 1
  • James J. Smith
    • 1
    • 3
    Email author
  1. 1.Department of EntomologyMichigan State UniversityE. LansingUSA
  2. 2.USDA Forest Service Northern Research StationE. LansingUSA
  3. 3.Lyman Briggs CollegeMichigan State UniversityE. LansingUSA
  4. 4.School of Agriculture and Consumer Sciences, Otis L. Floyd Nursery Research CenterTennessee State UniversityMcMinnvilleUSA

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