The ingestion of double-strand RNAs (dsRNA) targeting essential genes in an insect could cause mortality. Based on this principle, a new generation of insect control methods using RNA interference (RNAi) are being developed. In this work, we developed a bioassay for oral delivery of dsRNA to an invasive forest and urban tree pest, the emerald ash borer (EAB, Agrilus planipennis). EAB feeds and develops beneath the bark, killing trees rapidly. This behavior, coupled with the lack of a reliable artificial diet for rearing larvae and adults, make them difficult to study. We found that dsRNA is transported and processed to siRNAs by EAB larvae within 72 h after ingestion. Also, feeding neonate larvae with IAP (inhibitor of apoptosis) or COP (COPI coatomer, β subunit) dsRNA silenced their target genes and caused mortality. Both an increase in the concentration of dsRNA fed and sequential feeding of two different dsRNAs increased mortality. Here we provide evidence for successful RNAi in EAB, and demonstrate the development of a rapid and effective bioassay for oral delivery of dsRNA to screen additional genes.
Gall wasps (Hymenoptera: Cynipidae) induce structures (galls) on their host plants that house developing wasps and provide them with protection from natural enemies. The Asian chestnut gall wasp, Dryocosmus kuriphilus Yasumatsu, is an invasive pest that is destructive to chestnut (Castanea spp.). An improved understanding of the interactions among D. kuriphilus, its host, and its natural enemies is critical for the development of effective management strategies against this pest. The objective of our study was to evaluate the D. kuriphilus community interactions, and relate these interactions to variations among gall traits. Galls were collected from four locations throughout the eastern United States from May (gall initiation) through August (after gall wasp emergence), and January. Gall characteristics (volume, weight, and schlerenchyma layer thickness), gall inhabitants (D. kuriphilus, parasitoids, and chamber fungi), and other community associates (insect herbivores and lesions thought to be caused by endophytes) were evaluated and correlated using canonical correlation analyses. The primary mortality factors for D. kuriphilus were parasitism, gall chamber-invading fungi, and failure of adult gall wasps to emerge. Larger gall size and thicker schlerenchyma layers surrounding the larval chambers were negatively correlated with parasitoids and chamber fungi, indicating these gall traits are important defenses. External fungal lesions and insect herbivory were positively correlated with the absence of D. kuriphilus within galls. This study provides support for the protective role of cynipid galls for the gall inducer, identifies specific gall traits that influence gall wasp mortality, and improves our knowledge of D. kuriphilus ecology in North America.
Recent study has shown that RNA interference (RNAi) is efficient in emerald ash borer (EAB), Agrilus planipennis, and that ingestion of double-stranded RNA (dsRNA) targeting specific genes causes gene silencing and mortality in neonates. Here, we report on the identification of highly effective target genes for RNAi-mediated control of EAB. We screened 13 candidate genes in neonate larvae and selected the most effective target genes for further investigation, including their effect on EAB adults and on a non-target organism, Tribolium castaneum. The two most efficient target genes selected, hsp (heat shock 70-kDa protein cognate 3) and shi (shibire), caused up to 90% mortality of larvae and adults. In EAB eggs, larvae, and adults, the hsp is expressed at higher levels when compared to that of shi. Ingestion of dsHSP and dsSHI caused mortality in both neonate larvae and adults. Administration of a mixture of both dsRNAs worked better than either dsRNA by itself. In contrast, injection of EAB.dsHSP and EAB.dsSHI did not cause mortality in T. castaneum. Thus, the two genes identified cause high mortality in the EAB with no apparent phenotype effects in a non-target organism, the red flour beetle, and could be used in RNAi-mediated control of this invasive pest.
As the highly invasive hemlock woolly adelgid, Adelges tsugae, continues to expand its distribution in eastern North America, affected forests will incur drastic changes in composition and structure. While these changes have been wellstudied in dense hemlock forests in the Northeast, relatively little work is known about the effects of the adelgid at the western edge of the range of eastern hemlock, Tsuga canadensis. We evaluated the nature and extent of these changes using vegetation assessments coupled with growth simulations. The woody plant community was assessed in three strata (upper, mid-and lower) and was used to predict forest succession. Using the Forest Vegetation Simulator (FVS), we then projected the growth of hemlock forests 20 years into the future with and without the effects of the adelgid. In forest simulations lacking adelgid invasion, little change in composition or structure is forecast. In contrast, our projections predict a near complete loss of the hemlock forest type within 20 years of adelgid establishment, with widespread conversion to hardwood forest types, most notably white oak-red oak-hickory, chestnut oak-black oak-scarlet oak, and yellow poplar-white oak-red oak. Hemlock loss will result in denser deciduous forests with thinner canopies and multiple gaps, and significant alterations to terrestrial and aquatic wildlife habitat.
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