Nugent, Gregory D. and Coyne, Seamus and Nguyen, Thi T. and Kavanagh, Tony A. and Dix, Philip
(2006)
Nuclear and plastid transformation of Brassica oleracea var. botrytis (cauliflower) using PEG-mediated uptake of DNA into protoplasts.
Plant Science, 170 (1).
pp. 135-142.
ISSN 0168-9452
Abstract
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. Avector containing genes for hygromycin resistance and b-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 10ˉ⁵. No
spontaneous spectinomycin resistant mutants were observed in any plastid transformation experiments. PCR and Southern blot analysis confirmed
the transgenic status of plants regenerated from the protoplast-derived calli.
| Item Type: |
Article
|
| Keywords: |
Cauliflower; Plastid transformation; Nuclear transformation; Protoplast culture; |
| Academic Unit: |
Faculty of Science and Engineering > Biology |
| Item ID: |
7370 |
| Identification Number: |
https://doi.org/10.1016/j.plantsci.2005.08.020 |
| Depositing User: |
Prof. Philip J. Dix
|
| Date Deposited: |
23 Aug 2016 15:35 |
| Journal or Publication Title: |
Plant Science |
| Publisher: |
Elsevier |
| Refereed: |
Yes |
| URI: |
|
| Use Licence: |
This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available
here |
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