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

• Authors: S. Kumar , L. Arul , D. Talwar
• Languages of publication: EN

Abstracts

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.

Keywords

EN

BACILLUS THURINGIENSIS; MARKER-FREE TRANSGENIC; ORYZA SATIVA; SCIRPOPHAGA INCERTULAS; STEM BORER; TRANSGENIC RICE

Discipline

• GENETICS: GENETICS

• Publisher: Institute of Plant Genetics, Polish Academy of Sciences
• Journal: Journal of Applied Genetics
• Year: 2010
• Volume: 51
• Issue: 3
• Pages: 243-257

Contributors

author

S. Kumar

author

L. Arul

author

D. Talwar

• Document Type: ARTICLE
• Publication order reference: S. Kumar, Division of Crop Improvement, Indian Grassland and Fodder Research Institute, Jhansi-284 003, India