An environmental risk assessment of IPD079Ea: a protein derived from Ophioglossum pendulum with activity against Diabrotica spp.In maize.

Farmers in North America face significant pressure from insects in their maize fields, particularly from corn rootworm (Diabrotica spp.). Research into proteins capable of insecticidal activity has found several produced by ferns. One protein, IPD079Ea, was derived from Ophioglossum pendulum and has shown activity against corn rootworm. An environmental risk assessment was conducted for maize event DP-915635-4, which provides control of corn rootworms via expression of the IPD079Ea protein. This assessment focused on IPD079Ea and characterized potential exposure and hazard to non-target organisms (NTOs). For exposure, estimated environmental concentrations (EECs) were calculated. For hazard, laboratory dietary toxicity studies were conducted with IPD079Ea and surrogate non-target organisms. Environmental risk was characterized by comparing hazard and exposure to calculate the margin of exposure (MOE). Based on the MOE values for DP-915635-4 maize, the IPD079Ea protein is not expected to result in unreasonable adverse effects on beneficial NTO populations at environmentally relevant concentrations.

Evaluation of an in vitro experimental platform of human polarized intestinal epithelial monolayers for the hazard assessment of insecticidal proteins.

Previous work demonstrated the utility of using human-derived intestinal epithelial cell (IEC) lines cultured as polarized monolayers on Transwell® filters to differentiate between hazardous and non-hazardous proteins. The current study seeks to further resolve appropriate concentrations for evaluating proteins of unknown hazard potential using the IEC experimental platform and leverages these parameters for evaluating the potential toxicity of insecticidal proteins characteristic of those expressed in genetically modified (GM) agricultural biotechnology crops. To establish optimal test protein concentrations, effects of several known hazardous (C. perfringens epsilon toxin, Listeriolysin O, Phaseolus vulgaris erythroagglutinin, E. coli Shiga toxin 1, C. difficile Toxin B and wheat germ agglutinin) and non-hazardous (Ara-h2, ß-lactoglobulin, fibronectin and Rubisco) proteins on IEC barrier integrity and cell viability were evaluated at concentration ranges. Two insecticidal proteins (AfIP-1A and AfIP-1B) were evaluated for effects in the IEC assay, a seven-day insecticidal bioassay, and assessed in a high-dose 14-day acute oral toxicity study in mice. The results obtained from the human in vitro IEC assay were consistent with results obtained from an in vivo acute oral toxicity study, both demonstrating that the combination of AfIP-1A and AfIP-1B do not exhibit any identifiable harmful impacts on mammalian cells.

Characterization of the Spectrum of Activity of IPD079Ea: A Protein Derived From Ophioglossum pendulum (Ophioglossales: Ophioglossaceae) With Activity Against Western Corn Rootworm [Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)].

Western corn rootworm (Diabrotica virgifera virgifera LeConte) is a major pest of corn in both North America and Europe and as such presents significant challenges for farmers. IPD079Ea protein is encoded by the ipd079Ea gene from Ophioglossum pendulum (a species of fern) and was found to have activity against western corn rootworm in multiple corn events transformed to express the IPD079Ea protein. In chronic laboratory hazard studies, IPD079Ea protein was fed to eleven species in the order Coleoptera and four species in the order Lepidoptera to assess the spectrum of activity. Activity was observed on certain species of the Chrysomelidae and Coccinellidae families, with western corn rootworm as the most sensitive insect tested. No adverse effects on mortality or other sublethal endpoints were observed on any species within Lepidoptera. Overall, IPD079Ea protein appears not to have broad insecticidal properties and has potential value as an effective trait to control western corn rootworm in agricultural systems.