title: Development of broad-spectrum insect-resistant tobacco by expression of synthetic cry1Ac and cry2Ab genes.
abstract: Efficacy of two newly synthesized cry1Ac and cry2Ab genes was checked in tobacco before their expression in cotton. Both genes were artificially synthesized and codon optimized with respect to cotton-preferred codon usage. These genes were cloned in a plant expression vector and then transformed into tobacco. Fifty-eight putative transgenic plants were recovered from the selected explants. Successful integration of both genes in plant genome was confirmed by PCR amplification. Expression of transgenes was confirmed by PCR amplification from total plant RNA. Detached leaf insect bioassays were conducted with Helicoverpa armigera and Spodoptera exigua larvae. About 12 % of the transgenic plants showed significantly high resistance to S. exigua. Significant mortality (62 %) of H. armigera was recorded within 24 h of bioassays. Both toxins showed synergistic effect in tobacco and broadened the spectrum of plant activity against insects.
authors.affiliation: Plant Molecular Biology and Transformation Laboratory, Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Jhang Road, P.O. Box 577, Faisalabad, Pakistan.
authors.forename: Muhammad Nouman
authors.initials: MN
authors.lastname: Sohail
authors.forename: Sohail Mehmood
authors.initials: SM
authors.lastname: Karimi
authors.forename: Shaheen
authors.initials: S
authors.lastname: Asad
authors.forename: Shahid
authors.initials: S
authors.lastname: Mansoor
authors.forename: Yusuf
authors.initials: Y
authors.lastname: Zafar
authors.forename: Zahid
authors.initials: Z
authors.lastname: Mukhtar
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D004731:Endotoxins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D006460:Hemolysin Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D010572:Pest Control, Biological / Q000379:methods*; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D018411:Spodoptera / Q000187:drug effects; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis