Nugent, Gregory D. and ten Have, M. and van der Gulik, A. and Dix, Philip and Mordhorst, A.P. and Uijtewaal, B.A. (2005) Plastid transformants in tomato selected using mutations affecting ribosome structure. Plant Cell Reports, 24 (6). pp. 341-349. ISSN 0721-7714

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Abstract

Tomato plastid transformants were obtained using two vectors containing cloned plastid DNA of either Nicotiana tabacum or Solanum nigrum and including point mutations conferring resistance to spectinomycin and streptomycin. Transformants were recovered after PEG-mediated direct DNA uptake into protoplasts, followed by selection on spectinomycin-containing medium. Sixteen lines contained the point mutation, as confirmed by mapping restriction enzyme sites. One line obtained with each vector was analysed in more detail, in comparison with a spontaneous spectinomycin-resistant mutant. Integration of the cloned Solanum or Nicotiana plastid DNA, by multiple recombination events, into the tomato plastome was confirmed by sequence analysis of the targeted region of plastid DNA in the inverted repeat region. Maternal inheritance of spectinomycin and streptomycin resistances or sensitivity in seedlings also confirmed the transplastomic status of the two transformants. The results demonstrate the efficacy in tomato of a selection strategy which avoids the integration of a dominant bacterial antibiotic resistance gene.

Item Type:	Article
Keywords:	Tomato; Spectinomycin; Plastid transformation; Protoplasts;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7376
Identification Number:	https://doi.org/10.1007/s00299-005-0930-3
Depositing User:	Prof. Philip J. Dix
Date Deposited:	25 Aug 2016 16:01
Journal or Publication Title:	Plant Cell Reports
Publisher:	Springer
Refereed:	Yes
URI:	http://link.springer.com
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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