Occurrence and persistence of Bacillus thuringiensis (Bt) and transgenic Bt corn cry1Ab gene from an aquatic environment.
Genetically modified corn crops and suspensions of Bacillus thuringiensis (Bt) are currently used to control pest infestations of insects of the Lepidoptera family. For this purpose, the cry1Ab gene coding for protein delta-endotoxin derived from B. thuringiensis kurstaki (Btk), which is highly toxic to these insects, was inserted and expressed in corn. The aims of this study were to examine the occurrence and persistence of the cry1Ab gene from Btk and Bt corn in aquatic environments near fields where Bt corn was cultivated. First, an optimal DNA preparation and extraction methodology was developed to allow for quantitative gene analysis by real-time polymerase chain reaction (qPCR) in various environmental matrices. Second, surface water and sediment were spiked in vitro with genomic DNA from Bt or Bt corn to evaluate the persistence of cry1Ab genes. Third, soil, sediment, and water samples were collected before seeding, 2 weeks after pollen release, and after corn harvesting and mechanical root remixing in soils to assess cry1Ab gene content. DNA was extracted with sufficient purity (i.e., low absorbance at 230 nm and absence of PCR-inhibiting substances) from soil, sediment, and surface water. The cry1Ab gene persisted for more than 21 and 40 days in surface water and sediment, respectively. The removal of bacteria by filtration of surface water samples did not significantly increase the half-life of the transgene, but the levels were fivefold more abundant than those in unfiltered water at the end of the exposure period. In sediments, the cry1Ab gene from Bt corn was still detected after 40 days in clay- and sand-rich sediments. Field surveys revealed that the cry1Ab gene from transgenic corn and from naturally occurring Bt was more abundant in the sediment than in the surface water. The cry1Ab transgene was detected as far away as the Richelieu and St. Lawrence rivers (82 km downstream from the corn cultivation plot), suggesting that there were multiple sources of this gene and/or that it undergoes transport by the water column. Sediment-associated cry1Ab gene from Bt corn tended to decrease with distance from the Bt cornfield. Sediment concentrations of the cry1Ab gene were significantly correlated with those of the cry1Ab gene in surface water (R=0.83;P=0.04). The data indicate that DNA from Bt corn and Bt were persistent in aquatic environments and were detected in rivers draining farming areas.
Douville
M
M
Environment Canada, St. Lawrence Centre, 105 McGill Street, Montréal, Qué., Canada H2Y 2E7.
Gagné
F
F
Blaise
C
C
André
C
C
D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D018744:DNA, Plant / Q000032:analysis; D004731:Endotoxins / Q000235:genetics*; D004784:Environmental Monitoring; D019015:Geologic Sediments / Q000032:analysis; D006460:Hemolysin Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D012989:Soil Pollutants / Q000032:analysis; D019076:Transgenes; D014873:Water Pollutants / Q000032:analysis*; D003313:Zea mays / Q000235:genetics
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D018744:DNA, Plant; D004731:Endotoxins; D006460:Hemolysin Proteins; D012989:Soil Pollutants; D014873:Water Pollutants; C030956:insecticidal crystal protein, Bacillus Thuringiensis