Volatiles of High-Elevation Five-Needle Pines: Chemical Signatures through Ratios and Insight into Insect and Pathogen Resistance.

High-elevation five-needle pine trees are a group of Pinus species in the subgenus Strobus that occur at the edges of plant growth near the alpine tree line. These species are ecologically very important and are also threatened by climate-driven insect outbreaks and an exotic pathogen. Volatile organic compounds (VOCs) play central roles in the environmental adaptation of plants and in their defense against insects and pathogens. For example, the VOCs emitted by some high-elevation five-needle pine species attract female, tree-killing mountain pine beetles (MPB, Dendroctonus ponderosae) in the pioneering phase whereas VOCs from other species strongly repel this foremost herbivore, but the mechanism is unknown. We collected and compared headspace VOCs from foliage of eight species of high-elevation five-needle pines in Europe and North America. Overall, VOCs differed quantitatively among species with few qualitative differences. Despite species emitting essentially the same compounds, Random Forest analysis correctly classified 117 of the 126 trees sampled by using VOCs and identified the most important compounds for species classification and for separating species resistant from those susceptible to MPB or white pine blister rust (Cronartium ribicola). These VOC 'fingerprints' resulted largely from species emitting distinctive ratios of compounds, rather than through presence of species-specific compounds. Importantly, these Pinus species vary greatly in resistance to the main herbivore (MPB) and pathogen (white pine blister rust) causing tree mortality. Thus, these findings provide insights and should guide research into understanding resistance and in developing tools to manage these important trees. For instance, studies into the functions of five-needle pine VOCs in defense against abiotic or biotic stressors should focus on blend ratios rather than on individual compounds.

Mountain Pine Beetles Use Volatile Cues to Locate Host Limber Pine and Avoid Non-Host Great Basin Bristlecone Pine.

The tree-killing mountain pine beetle (Dendroctonus ponderosae Hopkins) is an important disturbance agent of western North American forests and recent outbreaks have affected tens of millions of hectares of trees. Most western North American pines (Pinus spp.) are hosts and are successfully attacked by mountain pine beetles whereas a handful of pine species are not suitable hosts and are rarely attacked. How pioneering females locate host trees is not well understood, with prevailing theory involving random landings and/or visual cues. Here we show that female mountain pine beetles orient toward volatile organic compounds (VOCs) from host limber pine (Pinus flexilis James) and away from VOCs of non-host Great Basin bristlecone pine (Pinus longaeva Bailey) in a Y-tube olfactometer. When presented with VOCs of both trees, females overwhelmingly choose limber pine over Great Basin bristlecone pine. Analysis of VOCs collected from co-occurring limber and Great Basin bristlecone pine trees revealed only a few quantitative differences. Noticeable differences included the monoterpenes 3-carene and D-limonene which were produced in greater amounts by host limber pine. We found no evidence that 3-carene is important for beetles when selecting trees, it was not attractive alone and its addition to Great Basin bristlecone pine VOCs did not alter female selection. However, addition of D-limonene to Great Basin bristlecone pine VOCs disrupted the ability of beetles to distinguish between tree species. When presented alone, D-limonene did not affect behavior, suggesting that the response is mediated by multiple compounds. A better understanding of host selection by mountain pine beetles could improve strategies for managing this important forest insect. Moreover, elucidating how Great Basin bristlecone pine escapes attack by mountain pine beetles could provide insight into mechanisms underlying the incredible longevity of this tree species.