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Transformation of peas
Authors:D R Davies  J Hamilton  P Mullineaux
Institution:(1) John Innes Centre for Plant Science Research, Colney Lane, NR4 7UH Norwich, UK
Abstract:Summary The lateral cotyledonary meristems present in germinating seed were inoculated with a non-oncogenic strain of A. tumefaciens carrying a gene conferring kanamycin resistance as a selectable marker and a beta-glucuronidase sequence as a reporter gene. Kanamycin resistant plants were derived from the meristems and shown to be transformed on the basis of Southern blots, polymerase chain reaction analysis and tests for beta-glucuronidase activity. The plants were fertile and tests of their progeny confirmed the transmission of integrated sequences through a sexual generation. This transformation method has the merit of an unlimited supply of material for inoculation and a relatively short time scale from inoculation to the production of rooted plants.Abbreviations BAP benzyl amino purine - GA3 gibberellic acid - GUS beta=glucuronidase - IBA indole butyric acid - MS Murashige and Skoog medium - NAA naphthalene acetic acid - NPTII neomycin phosphotransferase II - PCR polymerase chain reaction - TO first generation transformants - T1 second generation transformants
Keywords:Peas  Seed  Transformation  Agrobacterium  Meristems
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