Effects of tissue culture, biolistic transformation, and introduction of PPO and SPS gene constructs on performance of sugarcane clones in the field
J. E. Vickers A D , C. P. L. Grof A , G. D. Bonnett A E , P. A. Jackson B and T. E. Morgan CA CSIRO Plant Industry, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Qld 4067, Australia.
B CSIRO Plant Industry, Davies Laboratory, PMB, PO Aitkenvale, Qld 4814, Australia.
C CSR Technical Field Department, Kalamia Mill, PMB 6, Townsville, Qld 4810, Australia.
D Present address: Department of Biological and Physical Sciences, University of Southern Queensland, Toowoomba, Qld 4350, Australia.
E Corresponding author. Email: graham.bonnett@csiro.au
Australian Journal of Agricultural Research 56(1) 57-68 https://doi.org/10.1071/AR04159
Submitted: 5 July 2004 Accepted: 22 November 2004 Published: 31 January 2005
Abstract
Stably transformed sugarcane plants were produced by the biolistic introduction of DNA into tissue-cultured cells. Constructs containing genes in sense and antisense orientation of polyphenol oxidase and sense orientation of sucrose phosphate synthase were used in the transformations. Regenerated plants were grown in a series of field experiments that incorporated commercial varieties, including Q117, from which the transgenic clones were derived and plants regenerated from tissue culture but not subjected to biolistic bombardment. In all experiments, the mean yield of transgenic sugarcane was lower than commercial varieties and the transgenic clones often exhibited lower sugar content, although individual transgenic clones in some experiments were not significantly different from Q117. Those plants regenerated from tissue culture but not bombarded were intermediate in their yield, and more clones were equivalent to Q117 in agronomic performance. Transformed plants produced by the bombardment of callus performed poorly but the results from the tissue-cultured controls indicated that not all of this could be due to somaclonal variation. Some aspect(s) of the process of transformation itself was deleterious and in most cases more significant than the effects due to tissue culture. Of the transgenic clones grown at Ayr, Queensland, 1.6% were equivalent to Q117 in sugar content and yield, suggesting that large numbers of transgenic clones would have to be generated using the current method in order to allow for selection of clones with acceptable agronomic performance.
Acknowledgments
This work has been reliant upon a large number of colleagues, technical staff, and co-operators. We acknowledge the contributions of April Kartikasari, Scott Chapman, Peter Tuckett, John Foreman, Michael Hewitt, Bill Messer, Donna Glassop, Steve Attard, John Wilson, and John Manners (CSIRO), and Steve Elliot (CSR). BSES are thanked for supplying cane of commercial varieties for use in the experiments and for inspection of transgenics prior to transporting to north Queensland. We thank the Queensland Department of Primary Industries for their assistance with the experiment at Redland and, Nambour Mill for assistance in processing samples from the field. This research was partly funded by the CSIRO Tropical Agri-exports Multi-Divisional Program and the Sugar Research and Development Corporation.
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