Journal
PLANT SCIENCE
Volume 170, Issue 1, Pages 135-142Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2005.08.020
Keywords
cauliflower; plastid transformation; nuclear transformation; protoplast culture
Categories
Ask authors/readers for more resources
Most plastid transformation studies with tobacco, and all reports for other species (except tomato [G.D. Nugent, M. ten Have, A. van der Gulik, P.J. Dix, B.A. Uijtewaal, A.P. Mordhorst, Plastid transformants of tomato selected using mutations affecting ribosome structure. Plant Cell Rep. 24 (2005) 341-349]), have used biolistics for plastid transformation. However, nuclear transformation via biolistics has not been reported for any vegetable Brassica species so we used protoplast culture and PEG-mediated DNA uptake, to examine both nuclear and plastid transformation of cauliflower, an important vegetable Brassica. A vector containing genes for hygromycin resistance and beta-glucuronidase activity (pGUS-HYG) was used for nuclear transformation, while plastid transformation utilised a vector (pZB1) containing accD-rbcL plastome targeting regions cloned from Brassica napus (oil seed rape), and the selectable marker gene aadA, conferring resistance to spectinomycin. Protoplasts were embedded in agarose and selected on media containing hygromycin or spectinomycin. From five experiments, a single plastid transformant of the commercial cultivar Thalassa was obtained, whereas nuclear transformants were obtained at an absolute transformation frequency up to 1.3 x 10(-5). No spontaneous spectinomycin resistant mutants were observed in any plastid transformation experiments. PCR and Southern blot analysis confirmed the transgenic status of plants regenerated front the protoplast-derived calli. (c) 2005 Elsevier Ireland Ltd. All fights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available