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| Authors: | M.C. Christey, R.H. Braun, E.L. Conner, J.K. Reader, D.W.R. White, C.R. Voisey |
| Keywords: | Bt, Pieris rapae, Plutella xylostella, vegetable Brassicas, Agrobacterium |
Abstract:
Agrobacterium tumefaciens mediated transformation of Brassica oleracea vegetables and a rapid cycling B. oleracea line selected for high transformation ability was used to produce plants transgenic for a synthetic modified Bacillus thuringiensis (Bt) CRY1Ba1 or CRY1Ca5 gene.
Seedling explants were co-cultivated with A. tumefaciens strain LBA4404 containing a binary vector with an NPTII gene for selection of transgenic plants.
PCR, RT-PCR and ELISA analyses confirmed gene presence and expression.
Insect feeding assays were conducted using detached leaves of in vitro grown plants.
Neonate larvae of cabbage white butterfly (Pieris rapae) or diamond-back moth (Plutella xylostella) were placed on detached leaves.
Plants with high levels of gene expression caused mortality within 48 hrs.
Resistant plants were transferred to greenhouse conditions.
Insect assays were repeated on greenhouse-grown plants and confirmed high mortality of larvae on the transgenic plants.
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