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Recovery of morphologically normal transgenic tobacco from hairy roots co-transformed with Agrobacterium rhizogenes and a binary vector plasmid
Authors:H Hatamoto  M E Boulter  A H Shirsat  E J Croy  J R Ellis
Institution:(1) Department of Biological Sciences, University of Durham, South Road, DH1 3LE Durham, UK;(2) Present address: Technical Research Institute, Snow Brand Milk Products, 1-2, Minamidai 1-chome, Kawagoe, Saitama, Japan;(3) Present address: ICI Seeds, Jealott's Hill Research Station, RG12 6EY Bracknell, Berkshire, UK
Abstract:Summary Co-transformation of tobacco (Nicotiana tabacum) leaf explants with Agrobacterium rhizogenes harbouring pRi1855 and the binary vector pBin19 was achieved at a frequency of 67%. The kanamycin resistant hairy roots were cultured via a callusing phase to regenerate plants which were partially fertile when outcrossed with wild-type pollen. Phenotypic and molecular analysis of the F1 progeny demonstrated the efficient segregation of the hairy root marker from the kanamycin resistance marker, enabling morphologically normal plants to be recovered which retained the binary vector marker gene. This co-transformation strategy provides a means of introducing non-selectable genes into plants in cases where antibiotic resistance markers are undesirable.
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