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GrainGenes Reference Report: PRT-19-1188

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Reference	PRT-19-1188
Title	Transformation and expression of a gene for herbicide resistance in a Brazilian sugarcane
Journal	Plant Cell Reports
Year	2000
Volume	19
Pages	1188-1194
Author	Falco MC Neto AT Ulian EC
Abstract	Summary: 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
Keyword	[ Hide all but 1 of 34 ] agrobacterium-mediated transformation ammonium apparatus bar bombardment culture culture media cycle dna gene genotype herbicide resistance integration maize media microprojectile bombardment neomycin phosphotransferase nurseries particle bombardment phosphinotrycin acetyltransferase plasmid plasmids resistance resistant saccharum saccharum officinarum screenable marker stable integration stable transformation sugarcane transformation transgenic rice plants transgenic sugarcane western blotting

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