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PLANT PHYSIOLOGY , Vol 112, Issue 1 121-129, Copyright © 1996 by American Society of Plant Biologists
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PLANT-MICROBE AND PLANT-INSECT INTERACTIONS |
Genetic Transformation, Recovery, and Characterization of Fertile Soybean Transgenic for a Synthetic Bacillus thuringiensis cryIAc Gene
C. N. Stewart Jr, M. J. Adang, J. N. All, H. R. Boerma, G. Cardineau, D. Tucker and W. A. Parrott
Department of Crop and Soil Sciences (C.N.S., H.R.B., D.T., W.A.P.) and Department of Entomology (M.J.A., J.N.A.), The University of Georgia, Athens, Georgia 30602
Somatic embryos of Jack, a Glycine max (L.) Merrill cultivar, were
transformed using microprojectile bombardment with a synthetic Bacillus
thuringiensis insecticidal crystal protein gene (Bt cryIAc) driven by the
35S promoter and linked to the HPH gene. Approximately 10 g of tissue was
bombarded, and three transgenic lines were selected on
hygromycin-containing media and converted into plants. The recovered lines
contained the HPH gene, but the Bt gene was lost in one line. The plasmid
was rearranged in the second line, and the third line had two copies, one
of which was rearranged. The CryIAc protein accumulated up to 46 ng mg-1
extractable protein. In detached-leaf bioassays, plants with an intact copy
of the Bt gene, and to a lesser extent those with the rearranged copy, were
protected from damage from corn earworm (Helicoverpa zea), soybean looper
(Pseudoplusia includens), tobacco bud-worm (Heliothis virescens), and
velvetbean caterpillar (Anticarsia gemmatalis). Corn earworm produced less
than 3% defoliation on transgenic plants, compared with 20% on the
lepidopteran-resistant breeding line GatIR81-296, and more than 40% on
susceptible cultivars. Unlike previous reports of soybean transformation
using this technique, all plants were fertile. To our knowledge, this is
the first report of a soybean transgenic for a highly expressed
insecticidal gene.
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