Ephemeral wetlands are characterized by temporal changes in abiotic characteristics that could ameliorate or exacerbate contaminant effects on resident species. The goal of this study was to determine the effects of temperature and naturally occurring suspended solids and organic carbon on the response of Daphnia pulex, and the calanoid copepod, Diaptomus clavipes, to a copper reference toxicant. Organisms were exposed to copper at 10, 20 and 30 • C in 48-h static renewal tests with a diluent of either reconstituted laboratory water or water from a wetland that had elevated levels of both suspended solids and organic carbon. 48-h LC 50 values were calculated for both total and free ion copper concentrations. When wetland water was used as the diluent, LC 50 values based on total copper concentrations were significantly greater than free ion LC 50 s for both species. This difference was not as great in laboratory water, indicating that binding of the metal was greater in the wetland diluent and the free ion was largely responsible for toxicity. While D. clavipes was significantly less sensitive to the metal than D. pulex (48-h LC 50 for total copper in laboratory water at 20 • C 607.4 μg/L vs. 10.7 μg/L, respectively), the copepod exhibited a much greater response to increasing temperature. When the Biotic Ligand Model was used to generate free ion concentrations, it was found that measured concentrations exceeded the predicted values at each test condition; however measured LC 50 values for D. pulex were within a factor of two of the predicted LC 50 's at all temperatures and in both diluents.

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is considered one of the most economically important pests of maize (Zea mays L.) in the United States (U.S.) Corn Belt with costs of management and yield losses exceeding USD ~1–2 billion annually. WCR management has proven challenging given the ability of this insect to evolve resistance to multiple management strategies including synthetic insecticides, cultural practices, and plant-incorporated protectants, generating a constant need to develop new management tools. One of the most recent developments is maize expressing double-stranded hairpin RNA structures targeting housekeeping genes, which triggers an RNA interference (RNAi) response and eventually leads to insect death. Following the first description of in planta RNAi in 2007, traits targeting multiple genes have been explored. In June 2017, the U.S. Environmental Protection Agency approved the first in planta RNAi product against insects for commercial use. This product expresses a dsRNA targeting the WCR snf7 gene in combination with Bt proteins (Cry3Bb1 and Cry34Ab1/Cry35Ab1) to improve trait durability and will be introduced for commercial use in 2022.

Western corn rootworm ( Diabrotica virgifera virgifera LeConte) presents significant pest management challenges for farmers in both North America and Europe. IPD072Aa, a protein derived from Pseudomonas chlororaphis , has previously been shown to have activity against western corn rootworm. In the current study, the spectrum of activity of IPD072Aa was evaluated in controlled laboratory diet bioassays. IPD072Aa was fed at high concentrations in subchronic or chronic bioassays to 11 different insect species, representing 4 families within Coleoptera, and an additional 4 species representing four families of Lepidoptera. No adverse effects were noted in the Lepidoptera species. Within the order Coleoptera, western corn rootworm was the most sensitive species tested. A range of responses was observed within each of the four families of Coleoptera evaluated that included either no-observed effects or reduced growth, developmental delays, and/or reduced survival. These data will help inform the environmental risk assessment of genetically modified plants that express the IPD072Aa protein for western corn rootworm control.

Transgenic maize plants expressing dsRNA targeting western corn rootworm (WCR, Diabrotica virgifera virgifera) DvSSJ1 mRNA, a Drosophila snakeskin (ssk) ortholog, show insecticidal activity and significant plant protection from WCR damage. The gene encodes a membrane protein associated with the smooth sepate junction (SSJ) which is required for intestinal barrier function. To understand the active RNA form that leads to the mortality of WCR larvae by DvSSJ1 RNA interference (RNAi), we characterized transgenic plants expressing DvSSJ1 RNA transcripts targeting WCR DvSSJ1 mRNA. The expression of the silencing cassette results in the full-length transcript of 901 nucleotides containing a 210 bp inverted fragment of the DvSSJ1 gene, the formation of a double-stranded RNA (dsRNA) transcript and siRNAs in transgenic plants. Our artificial diet-feeding study indicates that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for DvSSJ1 insecticidal activity. Impact of specificity of dsRNA targeting DvSSJ1 mRNA on insecticidal activities was also evaluated in diet bioassay, which showed a single nucleotide mutation can have a significant impact or abolish diet activities against WCR. These results provide insights as to the functional forms of plant-delivered dsRNA for the protection of transgenic maize from WCR feeding damage and information contributing to the risk assessment of transgenic maize expressing insecticidal dsRNA.