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Transformation and regeneration of the self-incompatible species Nicotiana alata Link & Otto
Authors:Paul R Ebert  Adrienne E Clarke
Institution:(1) Plant Cell Biology Research Centre, School of Botany, University of Melbourne, 3052 Parkville, Victoria, Australia;(2) Present address: Department of Biology, Yale University, Kline Biology Tower, P.O. Box 6666, 06511-8112 New Haven, CT, USA
Abstract:A transformation and regeneration system has been developed for Nicotiana alata, a plant which is being intensively studied as a model of gametophytic self-incompatibility. Plantlets can be regenerated efficiently from seedling hypocotyls. Kanamycin-resistant, transformed plants have been obtained by cocultivation of regenerating hypocotyls with Agrobacterium tumefaciens strain LBA4404 containing a binary vector. The transformation frequency was low with <1% of tissue explants regenerating transformed plants. The transformed plants contained from one to three copies of the introduced DNA. In most cases, the kanamycin resistance phenotype was transmitted to the offspring as a normal Mendelian factor. In one unusual case, none of the offspring inherited the kanamycin resistance of the transformed maternal parent. This plant may have been chimeric or the kanamycin resistance gene may have been inactivated.
Keywords:Agrobacterium tumefaciens  hypocotyls  Nicotiana alata  regeneration  self-incompatibility  transformation
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