Generation and analysis of soybean plastid transformants expressing Bacillus thuringiensis Cry1Ab protoxin.
We describe the generation of fertile and homoplasmic soybean plastid transformants, expressing the Bacillus thuringiensis insecticidal protoxin Cry1Ab. Transgenes were targeted in the intergenic region of Glycine max plastome, between the rps12/7 and trnV genes and selection was carried out using the aadA gene encoding spectinomycin resistance. Molecular analysis confirmed the integration of the cry1Ab and aadA expression cassettes at the expected location in the soybean plastome, and the transmission of the transgenes to the next generation. Western blot analyses showed that the Cry1Ab protoxin is highly expressed in leaves, stems and seeds, but not in roots. Its expression confers strong insecticidal activity to the generated transgenic soybean, as exemplified with velvetbean caterpillar (Anticarsia gemmatalis).
Dufourmantel
N
N
Bayer BioScience, 14-20 rue Pierre Baizet, BP9163, 69263, Lyon Cedex 09, France, manuel.dubald1@bayercropscience.com.
Tissot
G
G
Goutorbe
F
F
Garçon
F
F
Muhr
C
C
Jansens
S
S
Pelissier
B
B
Peltier
G
G
Dubald
M
M
D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D001681:Biological Assay / Q000379:methods; D015139:Blotting, Southern; D015153:Blotting, Western; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics*; D006460:Hemolysin Proteins; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D007915:Lepidoptera / Q000187:drug effects / Q000254:growth & development; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D030821:Plants, Genetically Modified; D018087:Plastids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013025:Glycine max / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D014170:Transformation, Genetic / Q000235:genetics; D019076:Transgenes / Q000235:genetics
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D018744:DNA, Plant; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis