The instability of the flax element LIS-1 in transgenic Arabidopsis thaliana
Authors Bastaki N, Cullis C
Received 27 November 2014
Accepted for publication 3 February 2015
Published 14 May 2015 Volume 2015:6 Pages 89—105
Checked for plagiarism Yes
Review by Single-blind
Peer reviewer comments 4
Editor who approved publication: Professor Zvi Kelman
Nasmah K Bastaki, Christopher A Cullis
Department of Biology, Case Western Reserve University, Cleveland, OH, USA
Background: The LIS-1 is an element that appears as a site-specific insertion event in some flax lines in response to certain growth conditions and can be transmitted to subsequent generations. The origin of LIS-1 in the flax genome is uncertain. One possibility is that since LIS-1 does not exist intact in the progenitor line, it is assembled from small sequences found scattered throughout the genome, and that, under stressful growth conditions, induction occurs and these sequences are rearranged and assembled to form the intact LIS-1 element. It is unknown whether the intact LIS-1 element would remain stably integrated in other plant species or if it would be destabilized from their genome.
Results: In this study, Agrobacterium-mediated plant transformation via floral dipping was used to transform different accessions of the Columbia ecotype of Arabidopsis thaliana, with either LIS-1 or the target site into which LIS-1 integrates. The stability and the inheritance patterns of both elements were followed in subsequent generations. Our results indicate that, in the different transformed accessions, the target site of LIS-1 remains stable in the T1 and T2 generations. However, LIS-1 is not found intact in any transformed A. thaliana plants. Instead, it goes through multiple fragmentation events, which seem to be genotype dependent. In the process, the region originally flanking LIS-1 in the T-DNA construct can be converted to the same sequence found at the target site in flax, followed by complete excision of all the flax DNA in the construct.
Conclusion: These results demonstrate that the processes by which LIS-1 is produced in flax are also present in A. thaliana because both plants are capable of destabilizing the intact LIS-1 element.
Keywords: flax (Linum usitatissimum), Arabidopsis thaliana, plant transformation, Linum insertion sequence (LIS-1)
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