Expression of GFP and Bt transgenes in Brassica napus and hybridization with Brassica rapa |
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Authors: | M D Halfhill H A Richards S A Mabon C N Stewart |
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Institution: | (1) Department of Biology, University of North Carolina at Greensboro, Greensboro, NC 27402-6174, USA e-mail: nstewart@uncg.edu Tel.: 336-334-4980, Fax: 336-334-5839, US |
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Abstract: | It is possible to monitor the movement of transgenes by tagging them with green fluorescent protein (GFP). In order to develop
a model to study transgene flow, canola (Brassica napus cv Westar) was transformed with two GFP constructs, mGFP5er (GFP only) and pSAM 12 GFP linked to a synthetic Bacillus thuringiensis (Bt) cry1Ac endotoxin gene]. Transformed callus sectors that fluoresced green were preferentially selected in the tissue culture process.
Four independent GFP canola events and 12 events of GFP/Bt canola were regenerated through tissue culture. GFP fluorescence
was macroscopically detectable throughout the entire life cycle of canola. The GFP/Bt events were insecticidal to neonate
corn earworm (Helicoverpa zea) larvae and prevented herbivory damage. Fluorescence intensity at 508 nm varied between the independent transformation events,
and ranged from 7.6×105 to 13.8×105 (counts per second) in contrast with the wild-type at 5.3×105 cps. Nine GFP/Bt and three GFP events were hybridized with three wild accessions of B. rapa. The resultant hybrids fluoresced green and were insecticidal to neonate corn earworm larvae to the same degree as the transgenic
canola parents. However, fluorescence intensities of the hemizygous F1 hybrid lines were lower than the respective original homozygous canola parents. Each F1 hybrid line was backcrossed by hand onto the B. rapa parent, and transgenic backcrosses were produced at rates ranging from 15% to 34%. These data suggest that GFP can be used
as a tool to monitor transgene flow from crop species to wild relatives.
Received: 11 September 2000 / Accepted: 1 February 2001 |
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Keywords: | Transgenic canola GFP Interspecific hybridization Brassica rapa Bacillus thuringiensis |
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