Transformation and expression of a gene for herbicide resistance in a Brazilian sugarcane |
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Authors: | M C Falco A Tulmann Neto E C Ulian |
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Institution: | (1) Departamento de Genética ESALQ/USP. Caixa Postal 83, CEP 13418–900, Piracicaba, SP, Brazil e-mail: mcfalco@carpa.ciagri.usp.br, BR;(2) CENA/USP. Caixa Postal 97, CEP 13416–970, Piracicaba, SP, Brazil, BR;(3) COPERSUCAR. Caixa Postal 162, CEP 13400–970, Piracicaba, SP, Brazil, BR |
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Abstract: | Embryogenic calli of the Brazilian sugarcane (Saccharum officinarum L.) genotype SP80–180 were transformed with two plasmids containing genes coding for neomycin phosphotransferase (neo) and phosphinotrycin acetyltransferase (bar), by particle bombardment using an apparatus developed at Copersucar Technology Center. Transformed plants were initially
selected on culture medium containing Geneticin, and resistance was confirmed by localized application of a kanamycin solution
to leaves of hardened plants at the nursery. A commercial formulation of ammonium gluphosinate was sprayed twice on these
antibiotic-resistant plants. The resistant plants were considered co-transformed, and Southern analysis confirmed stable integration
of both bar and neo genes. In addition, phosphinotrycin acetyltransferase expression was supported by RT-PCR analysis and neomycin phosphotransferase
presence was demonstrated by western blotting. Similar analyses were also performed with micropropagated transformants after
three cycles of subculture.
Received: 15 December 1999 / Revision received: 15 April 2000 / Accepted: 1 June 2000 |
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Keywords: | Transgenic sugarcane Particle bombardment Neomycin phosphotransferase Phosphinotrycin acetyltransferase |
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