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GENOMICS, MOLECULAR GENETICS & BIOTECHNOLOGY

Characterization of Soybean Exhibiting High Expression of a Synthetic Bacillus thuringiensis cry1A Transgene That Confers a High Degree of Resistance to Lepidopteran Pests

^a Monsanto Co., 700 Chesterfield Village Pkwy. West, Chesterfield, MO 63017
^b Monsanto Co., 800 N. Lindbergh Blvd., Creve Coeur, MO 63167

^* Corresponding author (john.a.miklos{at}monsanto.com)

We report the generation of transgenic soybean [Glycine max (L.) Merr.] via Agrobacterium-mediated gene transfers of a cry1A gene (tic107) from Bacillus thuringiensis (Bt) that exhibits a high degree of resistance against the lepidopteran pests Pseudoplusia includens (Walker) (soybean looper), Helicoverpa zea (Boddie) (soybean podworm), and Anticarsia gemmatalis Hübner (velvetbean caterpillar). Three transgenic soybean lines (862, 726, and 781) were evaluated for expression, molecular composition, efficacy against target pests, and agronomic characteristics. We have designed and tested an expression cassette that consistently and reproducibly generates transgenic soybeans that accumulate the tic107 protein at levels as high as 6.12 µg mg^–1 of total extractable protein. Expression levels of this magnitude of a Bt insecticidal protein have never been reported in soybean. Previous reports have indicated expression levels 100-fold lower in the highest-expressing lines. In addition, the phenotypes of these high-expressing lines were indistinguishable from their negative segregant and the transformation parent (Asgrow var. ‘A3237’). Insect bioassay data demonstrate complete protection against soybean looper, soybean podworm, and velvetbean caterpillar when negative controls exhibited defoliation as high as 98%. Unlike previous reports of transgenic soybeans, we report here highly efficacious, single-copy, and normal phenotypes of transgenic soybean plants containing the highly expressed cry1A gene.

Abbreviations: Bt, Bacillus thuringiensis • PCR, polymerase chain reaction • RT, room temperature • T-DNA, transfer DNA