The maize Activator/Dissociation system is functional in hexaploid wheat through successive generations
Gabriela M. Pastori A , Alison Huttly A , Jevon West A , Caroline Sparks A , Alejandro Pieters B , Celina M. Luna C , Huw D. Jones A and Christine H. Foyer D EA Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK.
B IVI, Centro de Ecología, Altos de Pipe, Carretera Panamericana Km 11, Apartado 21827, Caracas 1020-A, Venezuela.
C Instituto de Fitopatología y FisiologíaVegetal (IFFIVE)-INTA, Camino 60 cuadras Km 51/2, 5009 Cordoba, Argentina.
D School of Agriculture, Food and Rural Development, Agriculture Building, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
E Corresponding author. Email: christine.foyer@newcastle.ac.uk
Functional Plant Biology 34(9) 835-843 https://doi.org/10.1071/FP07112
Submitted: 2 May 2007 Accepted: 3 July 2007 Published: 30 August 2007
Abstract
The aim of the present study was to provide useful background information and evidence of the functionality of the maize Activator/Dissociation (Ac/Ds) system in hexaploid wheat. Two transgenic parental wheat lines, one harbouring the immobilised Ac element (iAc) and the other the Ds element (pUbi[Ds-uidA]bar), were crossed. Transient GUS assays confirmed that the iAc transposase is active in hexaploid wheat. Selected F1 and F2 lines were analysed by PCR using primers specific to Ac, uidA and bar genes. The primer pair Ubi/bar-tag was used to detect excision of the Ds-uidA sequence, which occurred at a frequency of 39% in the F1 generation. Lines free of Ac and showing evidence of Ds excision were subject to Southern analysis, which indicated that at least one transposition event might have occurred in these lines. Although more evidence is required to unequivocally support the reintegration of the Ds element in the wheat genome, the evidence presented here nevertheless demonstrates the effectiveness and potential value of using this system to tag genes in wheat.
Acknowledgements
Rothamsted Research receives grant-aided support from the Biotechnology and Biological Sciences Research Council (BBSRC) of the United Kingdom. A. Pieters and C. M. Luna acknowledge short-term Royal Society fellowships. The authors thank Professor Peggy G. Lemaux (Berkeley University, USA) for the generous gift of constructs containing the maize Ac element and Act1(DsBar)uidA, Drs Sancha Salgueiro and Martin Cannell for helpful discussions and Nickerson’s Seeds for valuable advice on wheat crossings. The authors are grateful to Professors Narayana Upadhyaya, Bob Furbank, Tony Pryor, Mick Ayliffe (CSIRO, Canberra) and Professor Jonathan Jones (John Innes Centre, UK) for critical reading of the manuscript.
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