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

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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.

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Acknowledgments

We thank Dr. J. McLean for insightful discussion on MPB physiology and ecology, and Siew Law, Lynette Lim, Heather Jack, and Renee Morton for technical assistance. This work was supported by grants to CB and JB from the Natural Sciences and Engineering Research Council of Canada (NSERC) and funds to CB and JB for the Tria Project (www.thetriaproject.ca) provided by the Province of British Columbia through Genome Canada, Genome British Columbia and by the Government of Alberta through Genome Alberta.

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Correspondence to Colette Breuil.

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Cop. sp.1 described in this manuscript is probably the same as the one isolated in BC by Robinson (Bright and Stock 1982), we could not confirm this because, to our knowledge, the Robinson culture has not been preserved in a culture collection. Japanese C. minuta isolates from bark beetle galleries in spruce or pine have been shown to be non-pathogenic and to produce very small or no lesions in hosts (Lee et al. 2006). It is likely that Cop. sp.1 is not pathogenic to pine trees because it shows similar growth characteristics to the Japanese C. minuta, including, for example, slow growth, similar temperature requirements, cycloheximide sensitivity and mainly colonizing bark beetle galleries rather than phloem and sapwood. Compared to the MPB-associated staining fungi, Cop. sp.1 is difficult to detect using fungal isolation, since, on 1% MEA, this species grows at about 1mm/day in contrast to 25 mm/day for G. clavigera. Thus, Cop. sp.1 may have been considered unimportant in the MPB ecosystem, where fungal associates have been studied largely in relation to their effects on host trees (Yamaoka et al. 1995).

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Khadempour, L., LeMay, V., Jack, D. et al. The Relative Abundance of Mountain Pine Beetle Fungal Associates Through the Beetle Life Cycle in Pine Trees. Microb Ecol 64, 909–917 (2012). https://doi.org/10.1007/s00248-012-0077-z

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