A Metabolomic and HPLC-MS/MS Analysis of the Foliar Phenolics, Flavonoids and Coumarins of the Fraxinus Species Resistant and Susceptible to Emerald Ash Borer

The Emerald Ash Borer (EAB), Agrilus planipennis, Fairmaire, an Asian invasive alien buprestid has devastated tens of millions of ash trees (Fraxinus spp.) in North America. Foliar phytochemicals of the genus Fraxinus (Oleaceae): Fraxinus pennsylvanica (Green ash), F. americana (White ash), F. profunda (Bush) Bush. (Pumpkin ash), F. quadrangulata Michx. (Blue ash), F. nigra Marsh. (Black ash) and F. mandshurica (Manchurian ash) were investigated using HPLC-MS/MS and untargeted metabolomics. HPLC-MS/MS help identified 26 compounds, including phenolics, flavonoids and coumarins in varying amounts. Hydroxycoumarins, esculetin, esculin, fraxetin, fraxin, fraxidin and scopoletin were isolated from blue, black and Manchurian ashes. High-throughput metabolomics revealed 35 metabolites, including terpenes, secoiridoids and lignans. Metabolomic profiling indicated several upregulated putative compounds from Manchurian ash, especially fraxinol, ligstroside, oleuropin, matairesinol, pinoresinol glucoside, 8-hydroxypinoresinol-4-glucoside, verbenalin, hydroxytyrosol-1-O-glucoside, totarol and ar-artemisene. Further, dicyclomine, aphidicolin, parthenolide, famciclovir, ar-turmerone and myriocin were identified upregulated in blue ash. Principal component analysis demonstrated a clear separation between Manchurian and blue ashes from black, green, white and pumpkin ashes. The presence of defensive compounds upregulated in Manchurian ash, suggests their potential role in providing constitutive resistance to EAB, and reflects its co-evolutionary history with A. planipennis, where they appear to coexist in their native habitats.

Differential persistence of blue ash and white ash following emerald ash borer invasion

Catastrophic mortality of North American ash ( Fraxinus spp.) caused by Agrilus planipennis Fairmaire has been attributed to the lack of coevolved resistance between native ash species and this Asian invader. Although A. planipennis host preference or tree resistance can vary, all North American ash species are presumably highly vulnerable to A. planipennis. We inventoried live and dead blue ash ( Fraxinus quadrangulata Michx.) and white ash ( Fraxinus americana L.) in two southeastern Michigan woodlots several years after the A. planipennis invasion to assess their survival. Agrilus planipennis populations in this area peaked in approximately 2005, and the region is now characterized by nearly complete ash mortality. At the Plymouth site, 71% of the original 380 blue ash were alive, whereas only 29 saplings of the original 187 white ash were alive. At the Superior Township site, 63% of the original 210 blue ash were living, whereas all 125 white ash were dead. More than 80% of the blue ash had evidence of previous A. planipennis colonization, but 87% appeared healthy in 2011. Tree diameter did not consistently affect survival, and live and dead trees of both species were distributed across sites, indicating that differential survival was not attributable to localized conditions.

Emerald Ash Borer ( Agrilus planipennis) Density and Canopy Dieback in Three North American Ash Species

Emerald ash borer (Agrilus planipennis Fairmaire) (Coleoptera: Buprestidae), a phloem-feeding insect native to Asia, was identified in 2002 as the cause of widespread ash (Fraxinus) mortality in southeast Michigan, U.S. and Windsor, Ontario, Canada. Little information about A. planipennis is available from its native range and it was not known whether this invasive pest would exhibit a preference for a particular North American ash species. We monitored A. planipennis density and canopy condition on green ash (F. pennsylvanica) and white ash (F. americana) street trees in four neighborhoods and on white and blue ash (F. quadrangulata) trees in two woodlots in southeast Michigan. Green ash street trees had significantly more canopy dieback and higher A. planipennis densities than white ash trees growing in the same neighborhood. Density increased by two- to fourfold in both species over a 3-year period. Canopy dieback increased linearly from 2002 to 2005 as A. planipennis density increased (R 2= 0.70). In each of the woodlots, A. planipennis densities were significantly higher on white ash trees than blue ash trees. Woodpecker predation occurred in all sites and accounted for 35% of the A. planipennis that developed on trees we surveyed. Results indicate that surveys for A. planipennis detection in areas with multiple ash species should focus on the relatively preferred species.