, Volume 179, Issue 2, pp 467–485 | Cite as

Bacteria influence mountain pine beetle brood development through interactions with symbiotic and antagonistic fungi: implications for climate-driven host range expansion

  • Janet Therrien
  • Charles J. Mason
  • Jonathan A. Cale
  • Aaron Adams
  • Brian H. Aukema
  • Cameron R. Currie
  • Kenneth F. Raffa
  • Nadir ErbilginEmail author
Plant-microbe-animal interactions - Original research


Bark beetles are associated with diverse communities of symbionts. Although fungi have received significant attention, we know little about how bacteria, and in particular their interactions with fungi, affect bark beetle reproduction. We tested how interactions between four bacterial associates, two symbiotic fungi, and two opportunistic fungi affect performance of mountain pine beetles (Dendroctonus ponderosae) in host tissue. We compared beetle performance in phloem of its historical host, lodgepole pine (Pinus contorta), and its novel host recently accessed through warming climate, jack pine (Pinus banksiana). Overall, beetles produced more larvae, and established longer ovipositional and larval galleries in host tissue predominantly colonized by the symbiotic fungi, Grosmannia clavigera, or Ophiostoma montium than by the opportunistic colonizer Aspergillus and to a lesser extent, Trichoderma. This occurred in both historical and naïve hosts. Impacts of bacteria on beetle reproduction depended on particular fungus–bacterium combinations and host species. Some bacteria, e.g., Pseudomonas sp. D4–22 and Hy4T4 in P. contorta and Pseudomonas sp. Hy4T4 and Stenotrophomonas in P. banksiana, reduced antagonistic effects by Aspergillus and Trichoderma resulting in more larvae and longer ovipositional and larval galleries. These effects were not selective, as bacteria also reduced beneficial effects by symbionts in both host species. Interestingly, Bacillus enhanced antagonistic effects by Aspergillus in both hosts. These results demonstrate that bacteria influence brood development of bark beetles in host tissue. They also suggest that climate-driven range expansion of D. ponderosae through the boreal forest will not be significantly constrained by requirements of, or interactions among, its microbial associates.


Bark beetles Host colonization Invasion biology Bacteria Fungi Jack pine 



This project was funded by the USDA-Agriculture and Food Research Initiative (2003-3502-13528), and, in part, through post-graduate scholarships at the University of Alberta and from the Natural Sciences and Engineering Research Council of Canada—Discovery Grant, an Alberta Advanced Education and Technology Grant, and a Canada Research Chair Program awarded to NE. We thank Dr. Kathy Bleiker (Pacific Forestry Centre, Victoria, BC) for her valuable suggestions and feedback. Pam Melnick and Devon Letourneau from the Alberta Environment and Sustainable Resources Development helped us to set traps to collect live beetles. We thank two anonymous reviewers for helpful comments that improved our manuscript.


  1. Adams AS, Six DL (2007) Temporal variation in mycophagy and prevalence of fungi associated with developmental stages of Dendroctonus ponderosae (Coleoptera: Curculionidae). Environ Entomol 36:64–72CrossRefPubMedGoogle Scholar
  2. Adams AS, Currie CR, Cardoza Y, Klepzig KD, Raffa KF (2009) Effects of symbiotic bacteria and tree chemistry on the growth and reproduction of bark beetle fungal symbionts. Can J For Res 39:1133–1147CrossRefGoogle Scholar
  3. Adams AS, Jordan MS, Adams SM, Suen G, Goodwin LA, Davenport KW, Currie CR, Raffa KF (2011a) Community and genomic analysis of cellulose-degrading bacteria associated with the invasive woodwasp Sirex noctilio. Int Soc Microb Ecol 5:1323–1331Google Scholar
  4. Adams AS, Boone CK, Bohlmann J, Raffa KF (2011b) Responses of bark beetle-associated bacteria to host monoterpenes and their relationship to insect life history. J Chem Ecol 37:808–817CrossRefPubMedGoogle Scholar
  5. Adams AS, Aylward FO, Adams SM, Erbilgin N, Aukema BH, Currie CR, Suen G, Raffa KF (2013) Mountain pine beetles colonizing historical and naïve host trees are associated with a bacterial community highly enriched in genes contributing to terpene metabolism. Appl Environ Microbiol 79:3468–3475PubMedCentralCrossRefPubMedGoogle Scholar
  6. Addison AL, Powell JA, Six DL, Moore M, Bentz BJ (2013) The role of temperature variability in stabilizing the mountain pine beetle–fungus mutualism. J Theor Biol 335:40–50CrossRefPubMedGoogle Scholar
  7. Alamouti SM, Wang V, DiGuistini S, Six DL, Bohlmann J, Hamelin RC, Feau N, Breuil C (2011) Gene genealogies reveal cryptic species and host preferences for the pine fungal pathogen Grosmannia clavigera. Mol Ecol 20:2581–2602CrossRefPubMedGoogle Scholar
  8. Amman GD, Cole WE (1983) Mountain pine beetle dynamics in lodgepole pine forests. Part II: population dynamics. Gen. Tech. Report INT-145. USDA Forest Service, Intermountain Forest and Range Experiment Station, Ogden, UT, p 59Google Scholar
  9. Aukema BH, Raffa KF (2004) Behavior of adult and larval Platysoma cylindrica (Coleoptera: Histeridae) and larval Medetera bistriata (Diptera: Dolichopodidae) during subcortical predation of Ips pini (Coleoptera: Scolytidae). J Insect Behav 17:115–128CrossRefGoogle Scholar
  10. Aukema BH, Carroll AL, Zhu J, Raffa KF, Sickley TA, Taylor SW (2006) Landscape level analysis of mountain pine beetle in British Columbia, Canada: spatiotemporal development and spatial synchrony within the present outbreak. Ecography 29:427–441CrossRefGoogle Scholar
  11. Ayres M, Wilkens R, Ruel J, Lombardero M, Vallery E (2000) Nitrogen budgets of phloem-feeding bark beetles with and without symbiotic fungi. Ecology 81:2198–2210CrossRefGoogle Scholar
  12. Barras SJ (1970) Antagonism between Dendroctonus frontalis and the fungus Ceratocystis minor. Ann Entomol Soc Am 60:1187–1190CrossRefGoogle Scholar
  13. Barras SJ (1972) Improved White’s solution for surface sterilization of pupae of Dendroctonus frontalis. J Econ Entomol 65:1504CrossRefPubMedGoogle Scholar
  14. Bearup LA, Maxwell RM, Clow D, McCray JE (2014) Hydrological effects of forest transpiration loss in bark beetle-impacted watersheds. Nat Clim Change 4:481–486CrossRefGoogle Scholar
  15. Bentz B, Six DL (2006) Ergosterol content of fungi associated with Dendroctonus ponderosae and Dendroctonus rufipennis (Coleoptera: Curculionidae, Scolytinae). Ann Entomol Soc Am 99:189–194CrossRefGoogle Scholar
  16. Bentz BJ, Regniere J, Fettig CJ, Hansen EM, Hayes JL, Hicke JA, Kelsey RG, Negron JF, Seybold SJ (2010) Climate change and bark beetles of the western United States and Canada: direct and indirect effects. Bioscience 60:602–613CrossRefGoogle Scholar
  17. Bleiker KP, Six DL (2007) Dietary benefits of fungal associates to an eruptive herbivore: potential implications of multiple associates on host population dynamics. Environ Entomol 36:1384–1396CrossRefPubMedGoogle Scholar
  18. Blomquist GJ, Figueroa-Teran R, Aw M, Song M, Gorzalski A, Abbott NL, Chang E, Tittiger C (2010) Pheromone production in bark beetles. Insect Biochem Mol Biol 40:699–712CrossRefPubMedGoogle Scholar
  19. Boone CK, Keefover-Ring K, Mapes AC, Adams AS, Bohlmann J, Raffa KF (2013) Bacteria associated with a tree-killing insect reduce concentrations of plant defense compounds. J Chem Ecol 39:1003–1006CrossRefPubMedGoogle Scholar
  20. Brand JM, Bracke JW, Markovetz AJ, Wood DL, Browne LE (1975) Production of verbenol pheromone by a bacterium isolated from bark beetles. Nature 254:136–137CrossRefPubMedGoogle Scholar
  21. Bridges JR (1981) Nitrogen-fixing bacteria associated with bark beetles. Microb Ecol 7:131–137CrossRefPubMedGoogle Scholar
  22. Cardoza YJ, Klepzig KD, Raffa KF (2006) Bacteria in oral secretions of an endophytic insect inhibit antagonistic fungi. Ecol Entomol 31:636–645CrossRefGoogle Scholar
  23. Cardoza YJ, Vasanthakumar A, Suazo A, Raffa KF (2009) Survey and phylogenetic analysis of culturable microbes in the oral secretions of three bark beetle species. Entomol Exp Appl 131:138–147CrossRefGoogle Scholar
  24. Carmer SG, Swanson MR (1973) Evaluation of ten pairwise multiple comparison procedures by Monte Carlo methods. J Am Stat Assoc 68:66–74CrossRefGoogle Scholar
  25. Casteel CL, Hansen AK, Walling LL, Paine TD (2012) Manipulation of plant defense responses by the tomato psyllid (Bactericerca cockerelli) and its associated endosymbiont Candidatus Liberibacter psyllaurous. PLoS ONE 7(4):e35191. doi: 10.1371/journal.pone.0035191 PubMedCentralCrossRefPubMedGoogle Scholar
  26. Chung SH, Rosa C, Scully ED, Peiffer M, Tooker JJ, Hoover K, Luthe DS, Felton GW (2013) Herbivore exploits orally secreted bacteria to suppress plant defenses. Proc Natl Acad Sci USA 110:15728–15733PubMedCentralCrossRefPubMedGoogle Scholar
  27. Cigan PW, Karst J, Cahill JF Jr, Sywenky AN, Pec GJ, Erbilgin N (2015) Influence of bark beetle outbreaks on nutrient cycling in native pine stands in western Canada. Plant Soil 390:29–47CrossRefGoogle Scholar
  28. Clark EL, Pitt C, Carroll AL, Lindgren BS, Huber DPW (2014) Comparison of lodgepole and jack pine resin chemistry: implications for range expansion by the mountain pine beetle, Dendroctonus ponderosae (Coleoptera: Curculionidae). PeerJ 2:e240. doi: 10.7717/peerj.240 PubMedCentralCrossRefPubMedGoogle Scholar
  29. Colgan LJ, Erbilgin N (2011) Tree-mediated interactions between the jack pine budworm and a mountain pine beetle fungal associate. Ecol Entomol 36:425–434CrossRefGoogle Scholar
  30. Cudmore TJ, Björklund N, Carroll AL, Lindgren BS (2010) Climate change and range expansion of an aggressive bark beetle: evidence of higher beetle reproduction in naïve host tree populations. J Appl Ecol 47:1036–1043CrossRefGoogle Scholar
  31. DiGuistini S, Ralph SG, Lim YW, Holt R, Jones S, Bohlmann J, Breuil C (2007) Generation and annotation of lodgepole pine and oleoresin-induced expressed sequences from the blue-stain fungus Ophiostoma clavigerum, a mountain pine beetle-associated pathogen. FEMS Microbiol Lett 267:151–158CrossRefPubMedGoogle Scholar
  32. Dodds KJ, Graber C, Stephen FM (2001) Facultative intraguild predation by larval Cerambycidae (Coleoptera) on bark beetle larvae (Coleoptera: Scolytidae). Environ Entomol 30:17–22CrossRefGoogle Scholar
  33. Dowd PF, Shen SK (1990) The contribution of symbiotic yeast to toxin resistance of the cigarette beetle (Lasioderma serricorne F.). Entomol Exp Appl 56:241–248CrossRefGoogle Scholar
  34. Erbilgin N, Ma C, Whitehouse C, Shan B, Najar A, Evenden M (2014) Chemical similarity between historical and novel host plants promotes range and host expansion of the mountain pine beetle in a naïve host ecosystem. New Phytol 201:940–950CrossRefPubMedGoogle Scholar
  35. Godbout J, Beaulieu J, Bousquet J (2010) Phylogeographic structure of jack pine (Pinus banksiana; Pinaceae) supports the existence of a coastal glacial refugium in northeastern North America. Am J Bot 97:1903–1912CrossRefPubMedGoogle Scholar
  36. Goodsman DW, Erbilgin N, Lieffers VJ (2012) The impact of phloem nutrients on overwintering mountain pine beetles and their fungal symbionts. Environ Entomol 41:478–486CrossRefPubMedGoogle Scholar
  37. Gündüz AE, Douglas AE (2009) Symbiotic bacteria enable insect to use a nutritionally inadequate diet. Proc Biol Sci 276:987–991CrossRefGoogle Scholar
  38. Hofstetter TW, Moser JC (2014) The role of mites in insect–fungus associations. Annu Rev Entomol 59:537–557CrossRefPubMedGoogle Scholar
  39. Hulcr J, Adams A, Raffa KF, Hofstetter R, Klepzig K, Currie C (2011) Presence and diversity of Streptomyces in Dendroctonus bark beetle galleries across North America. Microb Ecol 61:759–768CrossRefPubMedGoogle Scholar
  40. Kaiser KE, McGlynn BL, Emanuel RE (2012) Ecohydrology of an outbreak: mountain pine beetle impacts trees in drier landscape positions first. Ecohydrology 6:444–454CrossRefGoogle Scholar
  41. Kikuchi Y, Hayatsu M, Hosokawa T, Nagayama A, Tago K, Fukatsu T (2012) Symbiont-mediated insecticide resistance. Proc Natl Acad Sci USA 109:8619–8622CrossRefGoogle Scholar
  42. Klepzig KD, Moser JC, Lombardero FL, Hofstetter RW, Ayres MPB (2001) Symbiosis and competition: complex interactions among beetles, fungi and mites. Symbiosis 30:83–96Google Scholar
  43. Kurz WA, Dymond CC, Stinson G, Rampley GJ, Neilson ET, Carroll AL, Ebata T, Safranyik L (2008) Mountain pine beetle and forest carbon: feedback to climate change. Nature 454:987–990CrossRefGoogle Scholar
  44. Langor DW, Raske AG (1987) Mortality factors and life tables of the eastern larch beetle, Dendroctonus simplex (Coleoptera: Scolytidae), in Newfoundland. Can Entomol 119:965–992Google Scholar
  45. Łukasik P, van Asch M, Guo H, Ferrari J, Godfray CJ (2013) Unrelated facultative endosymbionts protect aphids against a fungal pathogen. Ecol Lett 16:214–218CrossRefPubMedGoogle Scholar
  46. Mason CJ, Coutre JJ, Raffa KF (2014) Plant-associated bacteria degrade defense chemicals and reduce their adverse effects on an insect defoliator. Oecologia 175:901–910CrossRefPubMedGoogle Scholar
  47. Mason CJ, Rubert-Nason KF, Lindroth RL, Raffa KF (2015) Aspen defense chemicals influence midgut bacterial community composition of gypsy moth. J Chem Ecol 41:75–84CrossRefPubMedGoogle Scholar
  48. McGhehey JH (1971) Female size and egg production of the mountain pine beetle, Dendroctonus ponderosae Hopkins. Canadian Forest Service, Northern Forest Research Centre. Information Report NOR-X-9, p 18Google Scholar
  49. Morales-Jiménez J, Zúñiga G, Villa-Tanaca L, Hernández-Rodrígues C (2009) Bacterial community and nitrogen fixation in the red turpentine beetle, Dendroctonus valens, LeConte (Coleoptera: Curculionidae: Scolytinae). Microb Ecol 58:879–891CrossRefPubMedGoogle Scholar
  50. Morales-Jiménez J, Zúñiga G, Ramírez-Saad HC, Hernández-Rodríguez C (2012) Gut-associated bacteria throughout the life cycle of the bark beetle Dendroctonus rhizophagus Thomas and Bright (Curculionidae: Scolytinae) and their cellulolytic activities. Microb Ecol 64:268–278CrossRefPubMedGoogle Scholar
  51. Morales-Jiménez J, Vera-Ponce de León A, Garcia-Domínguez A, Martínez-Romero E, Zúñiga G, Hernández-Rodríguez G (2013) Nitrogen-fixing and uricolytic bacteria associated with the gut of Dendroctonus rhizophagus and Dendroctonus valens (Curculionidae: Scolytinae). Microb Ecol 66:200–210CrossRefPubMedGoogle Scholar
  52. Nardi JB, Mackie RI, Dawson JO (2002) Could microbial symbionts of arthropod guts contribute significantly to nitrogen fixation in terrestrial ecosystems? J Insect Physiol 48:751–763CrossRefPubMedGoogle Scholar
  53. Noda S, Kitade O, Inoue T, Kawai M, Kanuka M, Hiroshima K, Hongoh Y, Constantino R, Uys V, Zhong J, Kudo J, Ohkuma M (2007) Cospeciation in the triplex symbiosis of termite gut protists (Pseudotrichonympha spp.), their hosts, and their bacterial endosymbionts. Mol Ecol 16:1257–1266CrossRefPubMedGoogle Scholar
  54. Oliver KM, Russell JA, Moran NA, Hunter MS (2003) Facultative bacterial symbionts in aphids conifer resistance to parasitic wasps. Proc Natl Acad Sci USA 100:1803–1807PubMedCentralCrossRefPubMedGoogle Scholar
  55. Paine TD, Raffa KF, Harrington TC (1997) Interactions among scolytid bark beetles, their associated fungi, and live host conifers. Annu Rev Entomol 42:179–206CrossRefPubMedGoogle Scholar
  56. Parker BJ, Spragg CJ, Altincicek B, Gerardo NM (2013) Symbiont-mediated protection against fungal pathogens in pea aphids: a role for pathogen specificity? Appl Environ Microbiol 79:2455–2458PubMedCentralCrossRefPubMedGoogle Scholar
  57. Pec GJ, Karst J, Sywenky AN, Cigan PW, Erbilgin N, Simard SW, Cahill JF (2015) Rapid increases in forest understory diversity and productivity following a mountain pine beetle (Dendroctonus ponderosae) outbreak in pine forests. Plus One 10(4):e0124691CrossRefGoogle Scholar
  58. Poulsen M, Hu H, Li C, Chen Z, Xu L, Otani S et al (2014) Complementary symbiont contributions to plant decomposition in a fungus-farming termite. Proc Natl Acad Sci 111:14500–14505PubMedCentralCrossRefPubMedGoogle Scholar
  59. Raffa KF, Smalley EB (1995) Interactions of pre-attack and induced monoterpene concentrations in host conifer defense against bark beetle–microbial complexes. Oecologia 102:285–295CrossRefGoogle Scholar
  60. R Development Core Team (2012) R: a language and environment for statistical computing. R Foundation for Statistical Computing. Vienna, Austria. ISBN 3-900051-07-0.
  61. Reid RW, Whiney HS, Watson JA (1967) Reactions of lodgepole pine to attack by Dendroctonus ponderosae Hopkins and blue-stain fungi. Can J Bot 45:1115–1126CrossRefGoogle Scholar
  62. Rice AV, Thormann MN, Langor DW (2007) Virulence of, and interactions among, mountain pine beetle associated blue-stain fungi on two pine species and their hybrids in Alberta. Can J Bot 85:316–323CrossRefGoogle Scholar
  63. Romme WH, Knight DH, Yavitt JB (1986) Mountain pine beetle outbreaks in the Rocky Mountains: regulators of primary productivity? Am Nat 127:484–494CrossRefGoogle Scholar
  64. Safranyik L, Carroll AL (2006) The biology and epidemiology of the mountain pine beetle in lodgepole pine forests. In: Safranyik L, Wilson B (eds) The mountain pine beetle a synthesis of biology, management, and impacts on lodgepole pine. Canadian Forest Service, Pacific Forestry Center, Victoria, pp 3–66Google Scholar
  65. Safranyik L, Barclay H, Thomson A, Riel WG (1999) A population dynamics model for the mountain pine beetle, Dendroctonus ponderosae Hopk. (Coleoptera: Scolytidae). Inf. Rep. BC-X-386, Pac. For. Centre, Victoria, BCGoogle Scholar
  66. Safranyik L, Carroll AL, Regniere J, Langor DW, Riel WG, Shore TL, Peter B, Cooke BJ, Nealis VG, Taylor SW (2010) Potential for range expansion of mountain pine beetle into the boreal forest of North America. Can Entomol 142:415–442CrossRefGoogle Scholar
  67. Salom SM, Stephen FM, Thompson LC (1986) Development of Hylobius pales (Herbst) immatures in two types of phloem media. J Entomol Sci 21:43–51Google Scholar
  68. Schreiber LR, Gregory GF, Krause CR, Ichida JM (1988) Production, partial purification, and antimicrobial activity of a novel antibiotic produced by a Bacillus subtilis isolate from Ulmus americana. Can J Bot 66:2338–2346Google Scholar
  69. Scott JJ, Oh DC, Yuceer MC, Klepzig KD, Clardy J, Currie CR (2008) Bacterial protection of beetle–fungus mutualism. Science 322:63PubMedCentralCrossRefPubMedGoogle Scholar
  70. Six DL, Bentz BJ (2007) Temperature determine symbiont abundance in a multipartite bark beetle–fungus ectosymbiosis. Microb Ecol 54:112–118CrossRefPubMedGoogle Scholar
  71. Six DL, Paine TD (1998) Effects of mycangial fungi and host tree species on progeny survival and emergence of Dendroctonus ponderosae (Coleoptera: Scolytidae). Environ Entomol 27:1393–1401CrossRefGoogle Scholar
  72. Taft S, Najar A, Godbout J, Bousquet J, Erbilgin N (2015) Variation in foliar monoterpenes across the range of jack pine reveal three widespread chemotypes: implications to host expansion of invasive mountain pine beetle. Front Plant Sci 6:342. doi: 10.3389/fpls.2015.00342 PubMedCentralCrossRefPubMedGoogle Scholar
  73. Taylor AD, Hayes JL, Moser JC (1992) A phloem sandwich allowing attack and colonization by bark beetles (Coleoptera, Scolytidae) and associates. J Entomol Sci 27:311–316Google Scholar
  74. Thompson BM, Grebenok RJ, Behmer ST, Gruner DS (2013) Microbial symbionts shape the sterol Profile of the xylem-feeding woodwasp, Sirex noctilio. J Chem Ecol 39:129–139CrossRefPubMedGoogle Scholar
  75. Treu R, Karst J, Randall M, Pec GJ, Cigan P, Simard SW, Cooke J, Erbilgin N, Cahill JF (2014) Decline of ectomycorrhizal fungi following mountain pine beetle infestation. Ecology 95:1096–1103CrossRefPubMedGoogle Scholar
  76. Wood SL, Bright DE (1992) A catalog of Scolytidae and Platypodidae (Coleoptera), part 1 and 2. Taxonomic Index Volumes A and B. Great Basin Nat. Mem. 13A and B. Brigham Young Univ., Provo, UTGoogle Scholar
  77. Zilber-Rosenberg I, Rosenberg E (2008) Role of micro-organisms in the evolution of animals and plants: the hologenome theory of evolution. FEMS Microbiol Rev 32:723–735CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Janet Therrien
    • 1
  • Charles J. Mason
    • 2
  • Jonathan A. Cale
    • 1
  • Aaron Adams
    • 2
  • Brian H. Aukema
    • 3
  • Cameron R. Currie
    • 4
  • Kenneth F. Raffa
    • 2
  • Nadir Erbilgin
    • 1
    Email author
  1. 1.Department of Renewable ResourcesUniversity of AlbertaEdmontonUSA
  2. 2.Department of EntomologyUniversity of WisconsinMadisonUSA
  3. 3.Department of EntomologyUniversity of MinnesotaSt. PaulUSA
  4. 4.Department of BacteriologyUniversity of WisconsinMadisonUSA

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