Mountain pine beetle (Dendroctonus ponderosae) has killed millions of hectares of pine forests in western North America. Beetle success is dependent upon a community of symbiotic fungi comprised of Grosmannia clavigera, Ophiostoma montium, and Leptographium longiclavatum. Factors regulating the dynamics of this community during pine infection are largely unknown. However, fungal volatile organic compounds (FVOCs) help shape fungal interactions in model and agricultural systems and thus may be important drivers of interactions among bark beetle-associated fungi. We investigated whether FVOCs can mediate interspecific interactions among mountain pine beetle’s fungal symbionts by affecting fungal growth and reproduction. Headspace volatiles were collected and identified to determine species-specific volatile profiles. Interspecific effects of volatiles on fungal growth and conidia production were assessed by pairing physically-separated fungal cultures grown either on a carbon-poor or -rich substrate, inside a shared-headspace environment. Fungal VOC profiles differed by species and influenced the growth and/or conidia production of the other species. Further, our results showed that FVOCs can be used as carbon sources for fungi developing on carbon-poor substrates. This is the first report demonstrating that FVOCs can drive interactions among bark beetle fungal symbionts, and thus are important factors in beetle attack success.

Chemical induction can drive tree susceptibility to and host range expansions of attacking insects and fungi. Recently, mountain pine beetle (Dendroctonus ponderosae Hopkins; MPB) has expanded its host range from its historic host lodgepole pine (Pinus contorta var. latifolia Douglas ex Loudon) to jack pine (Pinus banksiana Lamb) in western Canada. Beetle success in jack pine forests likely depends upon the suitability of tree chemistry to MPB and its symbiotic phytopathogenic fungi. In particular, how rapid induced defenses of jack pine affect MPB colonization and the beetle's symbionts is unknown. In the field, we characterized and compared differences in rapid induced phloem monoterpenes between lodgepole and jack pines in response to various densities of Grosmannia clavigera (Robinson-Jeffery and Davidson)-a MPB symbiotic fungus used to simulate beetle attack-inoculations. Overall, lodgepole pine had higher limonene and myrcene, but lower α-pinene, concentrations than jack pine. However, myrcene concentrations in jack pine increased with inoculation density, while that in lodgepole pine did not respond to density treatments. We compared the growth and reproduction of MPB's symbiotic fungi, G. clavigera, Ophiostoma montium (Rumford) von Arx and Leptographium longiclavatum Lee, Kim and Breuil, grown on media amended with myrcene, α-pinene and limonene at concentrations reflecting two induction levels from each pine species. Myrcene and α-pinene amendments inhibited the growth but stimulated the reproduction of G. clavigera, whereas limonene stimulated its growth while inhibiting its reproduction. However, the growth and reproduction of the other fungi were generally stimulated by monoterpene amendments. Overall, our results suggest that jack pine rapid induction could promote MPB aggregation due to high levels of α-pinene (pheromone precursor), a positive feedback of myrcene (pheromone synergist) and low levels of limonene (resistance). Jack pine is likely as susceptible to MPB-vectored fungi as lodgepole pine, indicating that jack pine induction will likely not adversely affect symbiont activities enough to inhibit the invasion of MPB into jack pine forests.

Disturbances are frequent events across the Canadian boreal forest and can affect both below‐ and above‐ground ecosystem processes. How disturbances change below‐ground soil fungal communities and in‐turn affect pine establishment and performance is poorly understood. Such understanding has become increasingly important in light of observed changes in disturbance regimes in recent years due to climate change. We used a greenhouse experiment to determine how soil inoculum collected from lodgepole pine stands undisturbed (control) or disturbed by fire, mountain pine beetle outbreak, logging and salvage logging affect pine seedling performance in western Canada. We first characterized whether fungal communities of seedling roots change as a function of inoculum source, and then determined whether changes in fungal community composition impact pine seedling performance (biomass and height). Root fungal communities of pine seedlings from logged and salvage logged disturbances differed from their respective paired controls, while soils from natural disturbances (fire and beetle outbreak) did not. Among disturbances, the pine root fungal communities of fire and salvage logged disturbances differed. In parallel to the root fungal communities, seedling performance also decreased when comparing logging and salvage logging disturbances to paired controls. Among disturbance treatments, seedlings from the salvage logged disturbance did not grow as big as seedlings inoculated with soils from burned forests. Synthesis and application. Our findings indicate that anthropogenic disturbances (logging and salvage logging) can have cross‐generational impacts on pine seedling performance, through functional shifts in seedling root fungal community structure. Furthermore, the impacts of soil fungi on pine seedlings appear to be pronounced following salvage logging, stressing the importance of compound disturbance events. These findings may be important to land managers considering clear‐cut logging or salvage logging in pine forests, particularly where soil biotic communities are likely to be one of the predominate factors in pine establishment.

Plants interact simultaneously with multiple organisms, including ectomycorrhizal (EM) fungal symbionts which benefit plants by facilitating resource acquisition. Yet, their role in induced plant defenses that rely on the allocation of plant resources has received little attention. We investigated whether EM fungi can affect the induction of defense-related monoterpenes in greenhouse-grown lodgepole pine (Pinus contorta var. latifolia) seedlings, and whether such effects differed between EM fungal species occurring alone or in combination. Fungal interactions on growth media were also assessed to complement the greenhouse study. Our study revealed that the production of certain monoterpenes is influenced by the fungal species colonizing pine roots. Furthermore, pine seedlings did not necessarily benefit from having associations with multiple EM fungi, as we found contrasting effects of single vs. multiple species of fungi on induced monoterpene responses. Finally, monoterpene responses were altered when early-colonizing species inhibited the colonization or development of later-arriving species. We conclude that the presence of EM fungi can impact host susceptibility to insect and pathogen attack, suggesting that seedlings establishing in areas lacking fungi that promote the induction of tree defense chemicals may suffer from increased susceptibility to future pest damage.

Mountain pine beetle (MPB; Dendroctonus ponderosae Hopkins) has recently expanded its range into the lodgepole pine forests in Alberta, Canada. However, it is unknown whether semiochemical tools developed in the beetle’s historical range are suitable for monitoring MPB in the new environment. Thus, we conducted a 3-year study to test new MPB monitoring tools in Alberta. A field trial selected a combination of MPB pheromones and two host volatiles. Using this combination, we baited different numbers of trees in triangular, square, or rectangular formations (spatial arrangements of trees) to determine how the densities of baited trees affect MPB attraction. Three plots, each made up of three formations, were arranged in a linear transect at various distances between formation boundaries. The proportion of baited trees mass-attacked was highest in the square formation. However, the proportion of spillover trees mass-attacked (attacks on non-baited trees) was lower when formations were 1 km apart compared with 4 or 8 km apart. In a follow-up test of the square formation alone, there was no difference in trap tree effectiveness between distances of 8 and 12 km. We suggest that four baited trees spaced 50 m apart in a square formation at a 12 km distance can be used in the field.