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Protocols in ecological and environmental plant physiology


Article << Previous     |     Next >>   Contents Vol 56(1)

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, T. E. Morgan C

A 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
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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.

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