<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation of pineapple var. Queen using a novel encapsulation-based antibiotic selection technique |
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Authors: | Gaurab Gangopadhyay Subhash Kanti Roy Sangita Basu Gangopadhyay Kalyan Kumar Mukherjee |
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Institution: | (1) Cytogenetics Laboratory, Department of Botany, Bose Institute, 93/1 APC Road, Kolkata, 700009, West Bengal, India;(2) Department of Botany, Brahmananda Keshab Chandra College, 111/2 B.T. Road, Kolkata, 700108, West Bengal, India |
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Abstract: | A protocol for Agrobacterium-mediated transformation of a local ‘elite’ Indian variety (Queen) of pineapple Ananus comosus (L.) Merr, family Bromeliaceae] has been established using a standard transformation vector (pCAMBIA 1304). High transformation
efficiency, expressed as the mean percentage of transgenic micro-shoots regenerated from initial callus explants (20.6%) was
achieved using a novel encapsulation-based, antibiotic selection procedure. The Agrobacterium-infected micro-shoots derived from callus explants survived selection in high concentration of hygromycin (60 mg l−1 and beyond) in encapsulated alginate beads. The integration of transgene in hygromycin-resistant shoots and plants was confirmed
by histochemical GUS assay, PCR amplification and Southern hybridization. It is possible to eliminate false antibiotic positives
in pineapple transformation program to a large extent following this procedure. |
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Keywords: | Encapsulation-based selection Pineapple var Queen Transformation |
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