Ds transposition mediated by transient transposase expression in Heiracium aurantiacum |
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Authors: | Weld Richard J. Bicknell Ross A. Heinemann Jack A. Eady Colin C. |
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Affiliation: | (1) Plant and Microbial Sciences Department, University of Canterbury, Private Bag, Christchurch, New Zealand;(2) New Zealand Institute for Crop and Food Research, Private Bag 4704, Christchurch, New Zealand |
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Abstract: | The feasibility of using transient transposase expression to mobilize Ds elements for gene tagging in Hieracium aurantiacum was evaluated. A T-DNA construct carrying the Ac transposase gene and either a visible marker gene (uidA) or the conditionally-lethal marker gene (codA) was transferred to H. aurantiacum leaf discs (previously transformed with a Ds element) by co-cultivation with Agrobacterium tumefaciens. Shoots were regenerated directly from the co-cultivated leaf discs under selection for antibiotic resistance resulting from Ds excision. Most regenerants carried unique transposition events. Of 84 regenerated plants, twenty one (25%) did not express the marker gene and the DNA coding sequence of the transposase could not be detected in seven (8.3%). Potential advantages of this method over conventional gene-tagging methods are: rapid recovery of individual transposition events; regenerated plants are isogenic; and the transient nature of transposase expression should facilitate the stabilisation of the transposed element. |
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Keywords: | Ac/Ds gene tagging apomixis |
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