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1

The use of Bacillus thuringiensis in plant protection: prospects and limitations

100%
Malinowski H.
Biotechnologia
|
2000
|
issue 3
81-92
EN
Taking into account the protection of natural environment, it may be expected that the use of Bacillus thuringiensis toxins to control pest insects will be increased. It results from the discovery of novel toxins with different activity spectra and from the creation of more active toxins with the use of genetic engeeniering techniques. Besides, the introduction of transgenic plants such as cotton, potato, tomato, tobacco and others producing the bacterial toxins against more important species of pest insects will significantly reduce the application of chemical insecticides. The possibility of resistance development in insects poses a great threat to the above strategy. The better recognition of biochemical, physiological and genetical mechanisms of resistance to B. thuringiensis toxins will allow to devise a strategy for delaying jnsects resistance. The general principles of this strategy are similar to those used in the case of chemical insecticides and involve the rational application of B. thuringiensis toxins and their rotation with other insecticides.
2

Generation of marker-free Bt transgenic indica rice and evaluation of its yellow stem borer resistance

86%
Kumar S., Arul L., Talwar D.
Journal of Applied Genetics
|
2010
|
vol. 51
|
issue 3
243-257
EN
We report on generation of marker-free ('lean DNA') transgenic rice (Oryza sativa), carrying minimal gene-expression-cassettes of the genes of interest, and evaluation of its resistance to yellow stem borer Scirpophaga incertulas (Lepidoptera: Pyralidae). The transgenic indica rice harbours a translational fusion of 2 different Bacillus thuringiensis (Bt) genes, namely cry1B-1Aa, driven by the green-tissue-specific phosphoenol pyruvate carboxylase (PEPC) promoter. Mature seed-derived calli of an elite indica rice cultivar Pusa Basmati-1 were co-bombarded with gene-expression-cassettes (clean DNA fragments) of the Bt gene and the marker hpt gene, to generate marker-free transgenic rice plants. The clean DNA fragments for bombardment were obtained by restriction digestion and gel extraction. Through biolistic transformation, 67 independent transformants were generated. Transformation frequency reached 3.3%, and 81% of the transgenic plants were co-transformants. Stable integration of the Bt gene was confirmed, and the insert copy number was determined by Southern analysis. Western analysis and ELISA revealed a high level of Bt protein expression in transgenic plants. Progeny analysis confirmed stable inheritance of the Bt gene according to the Mendelian (3:1) ratio. Insect bioassays revealed complete protection of transgenic plants from yellow stem borer infestation. PCR analysis of T2 progeny plants resulted in the recovery of up to 4% marker-free transgenic rice plants.
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