Modifying fatty acid profiles through a new cytokinin-based plastid transformation system

A - Papers appearing in refereed journals

Dunne, A., Maple-Grodem, J., Gargano, D., Haslam, R. P., Napier, J. A., Chua, N-H., Russell, R. and Geir Moller, S. 2014. Modifying fatty acid profiles through a new cytokinin-based plastid transformation system. The Plant Journal. 80 (6), pp. 1131-1138.

AuthorsDunne, A., Maple-Grodem, J., Gargano, D., Haslam, R. P., Napier, J. A., Chua, N-H., Russell, R. and Geir Moller, S.
Abstract

The widespread use of herbicides and antibiotics for selection of transgenic plants has not been very successful with regard to commercialization and public acceptance. Hence, alternative selection systems are required. In this study, we describe the use of ipt, the bacterial gene encoding the enzyme isopentenyl transferase from Agrobacterium tumefaciens, as a positive selectable marker for plastid transformation. A comparison between the traditional spectinomycin-based aadA selection system and the ipt selection system demonstrated that selection of transplastomic plants on medium lacking cytokinin was as effective as selection on medium containing spectinomycin. Proof of principle was demonstrated by transformation of the kasIII gene encoding 3-ketoacyl acyl carrier protein synthaseIII into tobacco plastids. Transplastomic tobacco plants were readily obtained using the ipt selection system, and were phenotypically normal despite over-expression of isopentenyl transferase. Over-expression of KASIII resulted in a significant increase in 16:0 fatty acid levels, and a significant decrease in the levels of 18:0 and 18:1 fatty acids. Our study demonstrates use of a novel positive plastid transformation system that may be used for selection of transplastomic plants without affecting the expression of transgenes within the integrated vector cassette or the resulting activity of the encoded protein. This system has the potential to be applied to monocots, which are typically not amenable to traditional antibiotic-based selection systems, and may be used in combination with a negative selectable marker as part of a two-step selection system to obtain homoplasmic plant lines.

KeywordsPlant Sciences
Year of Publication2014
JournalThe Plant Journal
Journal citation80 (6), pp. 1131-1138
Digital Object Identifier (DOI)doi:10.1111/tpj.12684
PubMed ID25280363
Open accessPublished as bronze (free) open access
FunderNorwegian Research Council FORNY program
Plastid AS
Biotechnology and Biological Sciences Research Council
Funder project or codeDesigning Seeds for Nutrition and Health (DS)
*Design Oilseeds (Olga Sayanova)
Publisher's version
Output statusPublished
Publication dates
Online03 Oct 2014
Publication process dates
Accepted17 Sep 2014
PublisherWiley
Wiley-Blackwell
Copyright licensePublisher copyright
ISSN0960-7412

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