Microbial Ecology

, Volume 64, Issue 4, pp 909–917 | Cite as

The Relative Abundance of Mountain Pine Beetle Fungal Associates Through the Beetle Life Cycle in Pine Trees

  • Lily Khadempour
  • Valerie LeMay
  • David Jack
  • Jörg Bohlmann
  • Colette Breuil
Fungal Microbiology

Abstract

The mountain pine beetle (MPB) is a native bark beetle of western North America that attacks pine tree species, particularly lodgepole pine. It is closely associated with the ophiostomatoid ascomycetes Grosmannia clavigera, Leptographium longiclavatum, Ophiostoma montium, and Ceratocystiopsis sp.1, with which it is symbiotically associated. To develop a better understanding of interactions between beetles, fungi, and host trees, we used target-specific DNA primers with qPCR to assess the changes in fungal associate abundance over the stages of the MPB life cycle that occur in galleries under the bark of pine trees. Multivariate analysis of covariance identified statistically significant changes in the relative abundance of the fungi over the life cycle of the MPB. Univariate analysis of covariance identified a statistically significant increase in the abundance of Ceratocystiopsis sp.1 through the beetle life cycle, and pair-wise analysis showed that this increase occurs after the larval stage. In contrast, the abundance of O. montium and Leptographium species (G. clavigera, L. longiclavatum) did not change significantly through the MPB life cycle. From these results, the only fungus showing a significant increase in relative abundance has not been formally described and has been largely ignored by other MPB studies. Although our results were from only one site, in previous studies we have shown that the fungi described were all present in at least ten sites in British Columbia. We suggest that the role of Ceratocystiopsis sp.1 in the MPB system should be explored, particularly its potential as a source of nutrients for teneral adults.

Supplementary material

248_2012_77_MOESM1_ESM.jpg (48 kb)
Fig. 1(JPEG 48 kb)
248_2012_77_MOESM2_ESM.jpg (1.5 mb)
Fig. 2(JPEG 1522 kb)
248_2012_77_MOESM3_ESM.jpg (45 kb)
Fig. 3(JPEG 45 kb)

References

  1. 1.
    Bright D, Stock M (1982) Taxonomy and geographic variation. In: Mitton JB, Sturgeon KB (eds) Bark beetles in North American conifers: a system for the study of evolutionary biology. University of Texas Press, AustinGoogle Scholar
  2. 2.
    Six DL, Wingfield MJ (2011) The role of phytopathogenicity in bark beetle–fungus symbioses: a challenge to the classic paradigm. Annu Rev Entomol 56:255–272PubMedCrossRefGoogle Scholar
  3. 3.
    Harrington TC (2005) Ecology and evolution of mycophagous bark beetles and their fungal partners. In: Vega FE, Blackwell M (eds) Insect–fungal associations: ecology and evolution. Oxford University Press, New York, pp 257–291Google Scholar
  4. 4.
    BC MOF (2011) Mountain pine beetle. http://www.for.gov.bc.ca/hfp/mountain_pine_beetle/. Accessed 29 Oct 2011
  5. 5.
    Safranyik L, Carroll A (2007) The biology and epidemiology of the mountain pine beetle in lodgepole pine forests. In: The mountain pine beetle: a synthesis of biology, management and impacts on lodgepole pine. Natural Resources Canada, Pacific Forest Service, VictoriaGoogle Scholar
  6. 6.
    Whitney HS (1971) Association of Dendroctonus ponderosae (Coleoptera: Scolytidae) with blue stain fungi and yeasts during brood development in lodgepole pine. Can Entomol 103:1495–1503CrossRefGoogle Scholar
  7. 7.
    Bentz B, Logan J, Amman G (1991) Temperature-dependent development of the mountain pine-beetle (Coleoptera, Scolytidae) and simulation of its phenology. Can Entomol 123:1083–1094CrossRefGoogle Scholar
  8. 8.
    Paine TD, Raffa KF, Harrington T (1997) Interactions among scolytid bark beetles, their associated fungi, and live host conifers. Annu Rev Entomol 42:179–206PubMedCrossRefGoogle Scholar
  9. 9.
    Lee S, Kim J-J, Breuil C (2006) Diversity of fungi associated with the mountain pine beetle, Dendroctonus ponderosae and infested lodgepole pines in British Columbia. Fungal Divers 22:91–105Google Scholar
  10. 10.
    Yamaoka Y, Hiratsuka Y, Maruyama P (1995) The ability of Ophiostoma clavigerum to kill mature lodgepolepine trees. Eur J Forest Pathol 25:401–404CrossRefGoogle Scholar
  11. 11.
    Lee S, Kim J, Breuil C (2006) Pathogenicity of Leptographium longiclavatum associated with Dendroctonus ponderosae to Pinus contorta. Can J Forest Res 36:2864–2872CrossRefGoogle Scholar
  12. 12.
    Rumbold C (1941) A blue stain fungus, Ceratostomella montium n. sp., and some yeasts associated with two species of Dendroctonus. J Agric Res 62:589–601Google Scholar
  13. 13.
    Robinson R (1962) Blue stain fungi in lodgepole pine (Pinus contorta Dougl. var. latifolia Engelm.) infested by the mountain pine beetle (Dendroctonus monticolae Hopk.). Can J Bot 40:609–614CrossRefGoogle Scholar
  14. 14.
    Whitney HS, Farris SH (1970) Maxillary mycangium in mountain pine beetle. Science 167:54–55PubMedCrossRefGoogle Scholar
  15. 15.
    Six DL (2003) Bark beetle–fungus symbioses. In: Bourtzis K, Miller TA (eds) Insect symbiosis. CRC, Boca RatonGoogle Scholar
  16. 16.
    Six DL (2003) A comparison of mycangial and phoretic fungi of individual mountain pine beetles. Can J Forest Res 33:1331–1334CrossRefGoogle Scholar
  17. 17.
    Kim J, Allen E, Humble L, Breuil C (2005) Ophiostomatoid and basidiomycetous fungi associated with green, red, and grey lodgepole pines after mountain pine beetle (Dendroctonus ponderosae) infestation. Can J Forest Res 35:274–284CrossRefGoogle Scholar
  18. 18.
    Plattner A et al (2009) Resolving taxonomic and phylogenetic incongruence within species Ceratocystiopsis minuta. Mycologia 101:878–887PubMedCrossRefGoogle Scholar
  19. 19.
    Plattner A (2008) Pathogenicity and taxonomy of fungi associated with the mountain pine beetle in British Columbia. University of British Columbia, VancouverGoogle Scholar
  20. 20.
    Hsiau PTW, Harrington TC (2003) Phylogenetics and adaptations of basidiomycetous fungi fed upon by bark beetles (Coleoptera: Scolytidae). Symbiosis 34:111–131Google Scholar
  21. 21.
    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 27:1393–1401Google Scholar
  22. 22.
    Reid RW (1961) Moisture changes in lodgepole pine before and after attack by the mountain pine beetle. Forestry Chronicle 368–375Google Scholar
  23. 23.
    Clayton RB (1964) The utilization of sterols by insects. J Lipid Res 15:3–19PubMedGoogle Scholar
  24. 24.
    Norris DM, Baker JM, Chu HM (1969) Symbiotic interrelationships between microbes and ambrosia beetles. III. Ergosterol as the source of sterol to the insect. Ann Entomol Soc Am 62:413–414Google Scholar
  25. 25.
    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
  26. 26.
    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 36:1384–1396PubMedCrossRefGoogle Scholar
  27. 27.
    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
  28. 28.
    Adams AS, Six DL (2007) Temporal variation in mycophagy and prevalence of fungi associated with developmental stages of Dendroctonus ponderosae (Coleoptera: Curculionidae). Environ Entomol 36:64–72PubMedCrossRefGoogle Scholar
  29. 29.
    Luchi N, Capretti P, Pinzani P, Orlando C, Pazzagli M (2005) Real-time PCR detection of Biscogniauxia mediterranea in symptomless oak tissue. Lett Appl Microbiol 41:61–68PubMedCrossRefGoogle Scholar
  30. 30.
    Bahnweg G et al (2000) Controlled inoculation of Norway spruce (Picea abies) with Sirococcus conigenus: PCR-based quantification of the pathogen in host tissue and infection-related increase of phenolic metabolites. Trees-Struct Funct 14:435–441CrossRefGoogle Scholar
  31. 31.
    Schena L, Nigro F, Ippolito A, Gallitelli D (2004) Real-time quantitative PCR: a new technology to detect and study phytopathogenic and antagonistic fungi. Eur J Plant Pathol 110:893–908CrossRefGoogle Scholar
  32. 32.
    Karlsson M, Hietala A, Kvaalen H, Solheim H (2007) Quantification of host and pathogen DNA and RNA transcripts in the interaction of Norway spruce with Heterobasidion parviporum. Physiol Mol Plant Pathol 70:99–109CrossRefGoogle Scholar
  33. 33.
    Schweigkofler W et al (2005) Detection and quantification of Leptographium wageneri, the cause of black-stain root disease, from bark beetles (Coleoptera: Scolytidae) in Northern California using regular and real-time PCR. Can J For Res 35:1798–1808CrossRefGoogle Scholar
  34. 34.
    Carroll A, Taylor S, Régnière J, Safranyik L (2003) Effects of climate change on range expansion by the mountain pine beetle in British Columbia. In: Mountain pine beetle symposium: challenges and solutions Oct. 30–31. Kelowna, BCGoogle Scholar
  35. 35.
    Khadempour L, Massoumi Alamouti S, Hamelin RC, Bohlmann J, Breuil C (2010) Target-specific PCR primers can detect and differentiate ophiostomatoid fungi from microbial communities associated with the mountain pine beetle Dendroctonus ponderosae. Fungal Biol 114:825–833PubMedCrossRefGoogle Scholar
  36. 36.
    Cullings KW (1992) Design and testing of a plant-specific PCR primer for ecological and evolutionary studies. Mol Ecol 1:233–240CrossRefGoogle Scholar
  37. 37.
    West BT, Welch KB, Galecki AT (2006) Two-level models for clustered data: the rat pup example. In: West BT, Welch KB, Galecki AT (eds) Linear mixed models: a practical guide using statistical software. Chapman Hall/CRC, Boca Raton, p 376Google Scholar
  38. 38.
    Blom G (1958) Statistical estimates and transformed beta-variables. Wiley, New YorkGoogle Scholar
  39. 39.
    Holm S (1979) A simple sequentially rejective multiple test procedure. Scand J Stat 6:65–70Google Scholar
  40. 40.
    Raffa KF, Berryman A (1983) The role of host plant resistance in the colonization behavior and ecology of bark beetles (Coleoptera: Scolytidae). Ecological Monographs 53:27–49CrossRefGoogle Scholar
  41. 41.
    Bleiker KP, Six DL (2009) Competition and coexistence in a multi-partner mutualism: interactions between two fungal symbionts of the mountain pine beetle in beetle-attacked trees. Microb Ecol 57:191–202 http://www.springerlink.com/index/10.1007/s00248-008-9395-6.
  42. 42.
    Lee S (2006) Fungi associated with the mountain pine beetle, Dendroctonous ponderosae. University of British Columbia, VancouverGoogle Scholar
  43. 43.
    Upadhyay H (1981) A monograph of Ceratocystis and Ceratocystiopsis. University of Georgia Press, AthensGoogle Scholar
  44. 44.
    Gil-Serna J, González-Salgado A (2009) ITS-based detection and quantification of Aspergillus ochraceus and Aspergillus westerdijkiae in grapes and green coffee beans by real-time quantitative PCR. Int J Food Microbiol 131:162–167PubMedCrossRefGoogle Scholar
  45. 45.
    Suarez MB et al (2005) Development of real-time PCR (TaqMan) assays for the detection and quantification of Botrytis cinerea in planta. Plant Physiol Biochem 43:890–899PubMedCrossRefGoogle Scholar
  46. 46.
    Berbee ML, Taylor JW (1993) Dating the evolutionary radiations of the true fungi. Can J Bot 71:1114–1127CrossRefGoogle Scholar
  47. 47.
    Massoumi Alamouti S et al (2011) Gene genealogies reveal cryptic species and host preferences for the pine fungal pathogen Grosmannia clavigera. Mol Ecol 20:2581–2602CrossRefGoogle Scholar
  48. 48.
    Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986–994PubMedCrossRefGoogle Scholar
  49. 49.
    Gamper HA, Young JPW, Jones DL, Hodge A (2008) Real-time PCR and microscopy: are the two methods measuring the same unit of arbuscular mycorrhizal fungal abundance? Fungal Genet Biol 45:581–596PubMedCrossRefGoogle Scholar
  50. 50.
    Solheim H (1995) Early stages of blue-stain fungus invasion of lodgepole pine sapwood following mountain pine-beetle attack. Can J Bot 73:70–74CrossRefGoogle Scholar
  51. 51.
    Butler M, Day A (1998) Fungal melanins: a review. Can J Microbiol 44:1115–1136CrossRefGoogle Scholar
  52. 52.
    Six DL, Bentz BJ (2007) Temperature determines symbiont abundance in a multipartite bark beetle–fungus ectosymbiosis. Microb Ecol 54:112–118PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Lily Khadempour
    • 1
  • Valerie LeMay
    • 1
  • David Jack
    • 1
  • Jörg Bohlmann
    • 1
    • 2
  • Colette Breuil
    • 1
  1. 1.Faculty of ForestryUniversity of British ColumbiaVancouverCanada
  2. 2.Michael Smith LaboratoriesUniversity of British ColumbiaVancouverCanada

Personalised recommendations