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      Toxicological and biochemical analyses demonstrate no toxic effect of Cry1C and Cry2A to Folsomia candida

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          Abstract

          Collembolans are common soil arthropods that may be exposed to insecticidal proteins produced in genetically engineered (GE) plants by ingestion of crop residues or root exudates. In the present study, a dietary exposure assay was validated and used to assess the lethal and sublethal effects of two Bacillus thuringiensis ( Bt) insecticidal proteins, Cry1C and Cry2A, on Folsomia candida. Using the insecticidal compounds potassium arsenate (PA), protease inhibitor (E-64), and Galanthus nivalis agglutinin (GNA) mixed into Baker’s yeast, we show that the assay used can detect adverse effects on F. candida. Survival and development were significantly reduced when F. candida was fed a diet containing PA, E-64, and GNA at 9, 75, and 100 μg/g diet, respectively, but not when fed a diet containing 300 μg/g Cry1C or 600 μg/g Cry2A. The activities of test antioxidant-, detoxification-, and digestion-related enzymes in F. candida were unaltered by a diet containing 300 μg/g Cry1C or 600 μg/g Cry2A, but were significantly increased by a diet containing 75 μg/g E-64. The results confirm that Cry1C and Cry2A are not toxic to F. candida at concentrations that are much higher than those encountered under field conditions.

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          Assessment of risk of insect-resistant transgenic crops to nontarget arthropods.

          An international initiative is developing a scientifically rigorous approach to evaluate the potential risks to nontarget arthropods (NTAs) posed by insect-resistant, genetically modified (IRGM) crops. It adapts the tiered approach to risk assessment that is used internationally within regulatory toxicology and environmental sciences. The approach focuses on the formulation and testing of clearly stated risk hypotheses, making maximum use of available data and using formal decision guidelines to progress between testing stages (or tiers). It is intended to provide guidance to regulatory agencies that are currently developing their own NTA risk assessment guidelines for IRGM crops and to help harmonize regulatory requirements between different countries and different regions of the world.
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            Insecticidal activity of Bacillus thuringiensis crystal proteins.

            Published data on insecticidal activity of crystal proteins from Bacillus thuringiensis are incorporated into the Bt toxin specificity relational database. To date, 125 of the 174 holotype known toxins have been tested in approximately 1700 bioassays against 163 test species; 49 toxins have not been tested at all; 59 were tested against 71 Lepidoptera species in 1182 bioassays; 53 toxins were tested against 23 Diptera species in 233 bioassays; and 47 were tested against 39 Coleoptera species in 190 bioassays. Activity spectra of the tested toxins were summarized for each order. Comparisons of LC(50) values are confounded by high variability of the estimates, mostly due to within-species variation in susceptibility, and errors associated with estimation of toxin protein content. Limited analyses suggest that crystal protein toxicity is not affected by quarternary toxin rank or host used for gene expression, but that pre-ingestion treatment by solubilization or enzymatic processing has a large effect. There is an increasing number of toxin families with cross-order activity, as 15 of the 87 families (secondary rank) that are pesticidal are active against more than one order. Cross-order activity does not threaten environmental safety of B. thuringiensis-based pest control because toxins tend to be much less toxic to taxa outside the family's primary specificity range.
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              Recommendations for the design of laboratory studies on non-target arthropods for risk assessment of genetically engineered plants

              This paper provides recommendations on experimental design for early-tier laboratory studies used in risk assessments to evaluate potential adverse impacts of arthropod-resistant genetically engineered (GE) plants on non-target arthropods (NTAs). While we rely heavily on the currently used proteins from Bacillus thuringiensis (Bt) in this discussion, the concepts apply to other arthropod-active proteins. A risk may exist if the newly acquired trait of the GE plant has adverse effects on NTAs when they are exposed to the arthropod-active protein. Typically, the risk assessment follows a tiered approach that starts with laboratory studies under worst-case exposure conditions; such studies have a high ability to detect adverse effects on non-target species. Clear guidance on how such data are produced in laboratory studies assists the product developers and risk assessors. The studies should be reproducible and test clearly defined risk hypotheses. These properties contribute to the robustness of, and confidence in, environmental risk assessments for GE plants. Data from NTA studies, collected during the analysis phase of an environmental risk assessment, are critical to the outcome of the assessment and ultimately the decision taken by regulatory authorities on the release of a GE plant. Confidence in the results of early-tier laboratory studies is a precondition for the acceptance of data across regulatory jurisdictions and should encourage agencies to share useful information and thus avoid redundant testing.
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                Author and article information

                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group
                2045-2322
                23 October 2015
                2015
                : 5
                : 15619
                Affiliations
                [1 ]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences , Beijing 100193, China
                [2 ]College of Environment and Plant Protection, Hainan University , Haikou 570228, China
                [3 ]Qiongtai Teachers College , Haikou 571127, China
                [4 ]Agroscope, Institute for Sustainability Sciences ISS , 8046 Zurich, Switzerland
                Author notes
                [*]

                These authors contributed equally to this work.

                Article
                srep15619
                10.1038/srep15619
                4616050
                26494315
                e0778259-eba7-4a23-a400-714e2bc74ee1
                Copyright © 2015, Macmillan Publishers Limited

                This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

                History
                : 20 March 2015
                : 29 September 2015
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