Application of somatic embryogenesis for development of emerald ash borer-resistant white ash and green ash varietals.

Emerald ash borer (Agrilus planipennis; EAB) has devastated populations of ash (Fraxinus spp.) trees in dozens of U.S. states and Canada over the past few decades. The continued survival of scattered ash trees known as "lingering ash" in heavily infested natural stands, however, offers evidence of genetic resistance or tolerance to EAB. These surviving or "lingering" ash individuals may form the basis for reforestation programs in EAB-impacted areas, and clonal mass-propagation of these genotypes can help accelerate these efforts. Between 2013 and 2018, we initiated embryogenic cultures by culturing immature zygotic embryos from open-pollinated (OP) seeds collected from several surviving white ash and green ash trees in Michigan and Pennsylvania. In addition, in 2018, we initiated cultures from crosses made between lingering green ash parents from the USDA Forest Service ash breeding program in Ohio. Somatic embryos were produced by growing cultures in liquid suspension, followed by fractionation and plating on semisolid medium to produce developmentally synchronous populations of somatic embryos. Somatic embryo germination and conversion were enhanced by a combination of pre-germination cold treatment and inclusion of activated charcoal and gibberellic acid in the germination medium. Ash somatic seedlings derived from OP explants grew rapidly following transfer to potting mix and somatic seedlings representing nine ash clones were acclimatized, grown in the greenhouse and planted in a preliminary field test, along with EAB-resistant Manchurian ash (F. mandshurica) and EAB-susceptible control seedlings. Somatic seedlings have now been produced from cultures that originated from seeds derived from the progeny of lingering green ash parents and an ex vitro germination protocol has shown some promise for accelerating early somatic seedling growth. Results of this research could provide the basis for scaled-up production of EAB-resistant ash varieties for seed orchard production for forest restoration and cultivar development for urban tree restoration.

Potential for a minor pine bark beetle pest, Dendroctonus terebrans (Coleoptera: Curculionidae: Scolytinae), to mediate host location by a major pine killer, Dendroctonus frontalis.

The southern pine beetle, Dendroctonus frontalis Zimmermann is an important mortality agent of Pinus in the eastern United States of America where it commonly shares hosts with the black turpentine beetle, Dendroctonus terebrans (Olivier), which infrequently kills trees. Unlike D. frontalis, which must kill its hosts to become established in the bark and reproduce, D. terebrans can occupy living hosts as a parasite. Olfactory mechanisms whereby D. frontalis initially locates hosts have not been demonstrated, whereas D. terebrans responds strongly to host odors. Because D. terebrans produces frontalin, the primary aggregation pheromone component for D. frontalis, and commonly arrives on hosts prior to D. frontalis, it has been hypothesized that D. terebrans pheromone components can mediate D. frontalis location of suitable, living trees. We assessed this possibility with studies of the semiochemical interactions between D. frontalis and D. terebrans. Coupled gas chromatography-electroantennographic detection analyses indicated that D. terebrans produces nine different olfactory stimulants for D. frontalis, nearly all of them known semiochemicals for D. frontalis. A trapping experiment designed to address the potentially confounding influence of lure contamination confirmed that the D. terebrans pheromone component exo-brevicomin enhances attraction of D. frontalis and thus could be an attractive kairomone. In ambulatory bioassays, male D. frontalis were strongly attracted to odors of frass of solitary female and paired D. terebrans, indicating their attraction to the naturally occurring semiochemicals of D. terebrans. Cues from D. terebrans may influence host and mate-finding success of D. frontalis and, thereby, the latter's virulence.

Attaching lures to multiple-funnel traps targeting saproxylic beetles (Coleoptera) in pine stands: inside or outside funnels?

We conducted two field trapping experiments with multiple-funnel traps in 2008 and one experiment in 2010 to determine the effects of lure placement (inside or outside funnels) on catches of saproxylic species of beetles (Coleoptera). The experiments were conducted in southern pine (Pinus spp.) stands in central Georgia using combinations of ethanol, alpha-pinene, ipsenol, and ipsdienol lures. We report on a modification to the multiple-funnel trap that allows placement of large lures inside the confines of the funnels with minimal blockage. In general, catches of five species of common longhorn beetles (Cerambycidae), two species of regeneration weevils (Curculionidae), four species of bark beetles (Curculionidae: Scolytinae), and seven species of beetle predators and ectoparasites (Cleridae, Histeridae, Tenebrionidae, Trogossitidae, and Zopheridae) were higher in funnel traps with lures attached inside the funnels than in those with lures attached outside of the funnels. Catches of the remaining species were unaffected by lure placement. In no instance were catches of any species lower in funnel traps with lures attached inside the funnels than in those with lures attached outside of the funnels. For most species, catches in modified funnel traps with ethanol, alpha-pinene, ipsenol, and ipsdienol lures attached inside funnels were comparable with those in cross-vane panel traps.

Bioaccumulation of contaminants in Scarabaeidae and Silphidae beetles at sites polluted by coal combustion residuals and radiocesium.

Anthropogenic contamination from coal-fired power plants and nuclear reactors is a pervasive issue impacting ecosystems across the globe. As a result, it is critical that studies continue to assess the accumulation and effects of trace elements and radionuclides in a diversity of biota. In particular, bioindicator species are a powerful tool for risk assessment of chemically contaminated habitats. Using inductively coupled plasma mass spectrometry (ICP-MS) and auto-gamma counting, we analyzed trace element and radiocesium contaminant concentrations in Scarabaeidae and Silphidae beetles (Order: Coleoptera), important taxa in decomposition and nutrient cycling, at contaminated and reference sites on the Savannah River Site, South Carolina, U.S. Our results revealed variability in trace element concentrations between Scarabaeidae and Silphidae beetles at uncontaminated and contaminated sites. Compared to Scarabaeidae, Silphidae had higher levels of chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), and zinc (Zn). Unexpectedly, concentrations of Cr, Cu, and Ni were higher in both taxa at the uncontaminated sites. Scarabaeidae and Silphidae beetles at the coal combustion waste site consistently had high concentrations of arsenic (As), and Scarabaeidae had high concentrations of selenium (Se). Of the 50 beetles analyzed for radiocesium levels, two had elevated radioactivity concentrations, both of which were from a site contaminated with radionuclides. Our results suggest carrion beetles may be particularly sensitive to bioaccumulation of contaminants due to their trophic position and role in decomposition, and thus are useful sentinels of trace element and radionuclide contamination.

Effects of industrial disturbances on biodiversity of carrion-associated beetles.

Energy production systems such as nuclear reactors and coal-burning power plants produce a multitude of waste contaminants including radionuclides, trace elements, and heavy metals. Among invertebrates, much of the effort to understand the impact of these contaminants has focused in aquatic environments, while relatively less attention has been on terrestrial communities. We investigated the effects of trace element and radionuclide contamination on assemblages of beetles that are drawn to vertebrate carrion. Samples were collected from riparian sites at the Savannah River Site in South Carolina to compare trap catches (i.e., measure of relative abundance) of beetles and species diversity along a habitat gradient (0-300 m) away from an aquatic habitat and between uncontaminated and contaminated sites. We collected 17,800 carrion-associated beetles representing 112 species in nine families, which were classified as either scavenger or predatory beetles. Beetle catches and species diversity were generally higher at contaminated than uncontaminated sites. These trends were likely driven by scavenger species, which showed similar patterns between sites, whereas patterns of catches and species diversity were variable between sites for predatory beetles. Species compositions of contaminated and uncontaminated sites were generally distinct, however habitat edges appeared to substantially affect beetle assemblages. Overall, our study suggests carrion beetle assemblages are sensitive to edge effects and may exhibit variable responses to the presence of anthropogenic contaminants or disturbances associated with energy production systems. Such results reflect the inherent variability among individual beetle species, populations, and communities to local environmental conditions, and underscores the need for multi-taxa approach in environmental impact assessments.