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Cry3Bb1 protein from Bacillus thuringiensis in root exudates and biomass of transgenic corn does not persist in soil

Abstract

The Cry3Bb1 protein, insecticidal to the corn rootworm complex (Diabrotica spp.), of Bacillus thuringiensis (Bt) subsp. kumamotoensis was released in root exudates of transgenic Bt corn (event MON863) in sterile hydroponic culture (7.5 ± 1.12 ng/ml after 28 days of growth) and in nonsterile soil throughout growth of the plants (2.2 ± 0.62 ng/g after 63 days of growth). Kitchawan soil, which contains predominantly kaolinite (K) but not montmorillonite (M), was amended to 3 or 6% (vol./vol.) with K (3K and 6K soils) or M (3M and 6M soils) and with 1, 3, 5, or 10% (wt./wt.) of ground biomass of Bt corn expressing the Cry3Bb1 protein and incubated at 25 ± 2°C at the –33-kPa water tension for 60 days. Soils were analyzed for the presence of the protein every 7 to 10 days with a western blot assay (ImmunoStrip) and verified by ELISA. Persistence of the protein varied with the type and amount of clay mineral and the pH of the soils and increased as the concentration of K was increased but decreased as the concentration of M was increased. Persistence decreased when the pH of the K-amended soils was increased from ca. 5 to ca. 7 with CaCO3: the protein was not detected after 14 and 21 days in the pH-adjusted 3K and 6K soils, respectively, whereas it was detected after 40 days in the 3K and 6K soils not adjusted to pH 7. The protein was detected for only 21 days in the 3M soil and for 14 days in the 6M soil, which were not adjusted in pH. These results indicate that the Cry3Bb1 protein does not persist or accumulate in soil and is degraded rapidly.

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Acknowledgments

These studies were supported, in part, by grant 2003-35107-13776 from the US Department of Agriculture and grant N5238 from the NYU Research Challenge Fund. The opinions expressed herein are not necessarily those of the granting agencies. We thank the Monsanto Company for providing the corn seeds and purified Cry3Bb1 protein.

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Correspondence to Guenther Stotzky.

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Icoz, I., Stotzky, G. Cry3Bb1 protein from Bacillus thuringiensis in root exudates and biomass of transgenic corn does not persist in soil. Transgenic Res 17, 609–620 (2008). https://doi.org/10.1007/s11248-007-9133-8

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  • DOI: https://doi.org/10.1007/s11248-007-9133-8

Keywords

  • Bacillus thuringiensis
  • Transgenic corn
  • Cry3Bb1 protein
  • Corn rootworm
  • Root exudates
  • Clay minerals
  • pH
  • Persistence