title: Genetic transformation, recovery, and characterization of fertile soybean transgenic for a synthetic Bacillus thuringiensis cryIAc gene.
abstract: 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 rear-ranged. 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 budworm (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.
authors.affiliation: Department of Crop and Soil Sciences, University of Georgia, Athens 30602, USA.
authors.forename: C N
authors.initials: CN
authors.lastname: Stewart
authors.forename: M J
authors.initials: MJ
authors.lastname: Adang
authors.forename: J N
authors.initials: JN
authors.lastname: All
authors.forename: H R
authors.initials: HR
authors.lastname: Boerma
authors.forename: G
authors.initials: G
authors.lastname: Cardineau
authors.forename: D
authors.initials: D
authors.lastname: Tucker
authors.forename: W A
authors.initials: WA
authors.lastname: Parrott
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity; D001427:Bacterial Toxins; D001681:Biological Assay; D017931:DNA Primers; D004731:Endotoxins / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity; D005798:Genes, Bacterial; D005813:Genes, Synthetic; D006460:Hemolysin Proteins; D007915:Lepidoptera; D010572:Pest Control, Biological; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D013025:Glycine max / Q000235:genetics / Q000502:physiology*; D014170:Transformation, Genetic
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004731:Endotoxins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase