The N-terminal presequence from F1-ATPase β-subunit of Nicotiana plumbaginifolia efficiently targets green fluorescent fusion protein to the mitochondria in diverse commercial crops
Annathurai Gnanasambandam A B D , David J. Anderson A B , Matthew P. Purnell A B , Lars K. Nielsen B C and Stevens M. Brumbley A B C DA BSES Limited, 50 Meiers Road, Indooroopilly, Qld 4068, Australia.
B Cooperative Research Centre for Sugar Industry Innovation through Biotechnology, The University of Queensland, St Lucia, Qld 4072, Australia.
C Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, Qld 4072, Australia.
D Corresponding authors. Emails: agnanasambandam@bses.org.au; s.brumbley1@uq.edu.au
Functional Plant Biology 35(2) 166-170 https://doi.org/10.1071/FP07277
Submitted: 22 November 2007 Accepted: 8 February 2008 Published: 19 March 2008
Abstract
Approximately 10–15% of plant nuclear genes appear to encode mitochondrial proteins that are directed to mitochondria by specific targeting signals. Reports on the heterologous function of these targeting signals are generally limited to one or a few species, with an emphasis on model plants such as tobacco and Arabidopsis. Given their sequence diversity and their insufficient testing in commercially important crops (including monocotyledonous crops), the extent to which these signals can be relied on for biotechnological purposes across species remains to be established. This study provides the experimental verification of a mitochondrial signal that is functional across diverse crop species, including five monocots (sugarcane, wheat, corn, sorghum and onion) and seven dicots (cucumber, cauliflower, tomato, capsicum, pumpkin, coriander and sunflower). In all 12 crops, transient assays following microprojectile bombardment showed that the N-terminal mitochondrial presequence from F1-ATPase β-subunit (ATPase-β) of Nicotiana plumbaginifolia Viv. targeted green fluorescent fusion protein to the mitochondria. The transient assay results in sugarcane were confirmed in stably transformed root cells. The ATPase-β signal should be a useful metabolic engineering tool for directing recombinant proteins to the mitochondrial matrix in diverse plant species of commercial interest.
Additional keywords: mitochondrial presequence, protein targeting, confocal laser scanning microscopy.
Acknowledgements
A. Gnanasambandam is a recipient of a ‘Smart State Fellowship’ awarded by the Department of State Development, Trade and Innovation of the Queensland Government. This work was funded by the Cooperative Research Centre for Sugar Industry Innovation through Biotechnology. The authors thank Ms Edwina Williams for excellent technical assistance, Dr Agnelo Furtado for providing pUBI-GFP, Dr Itsara Pracharoenwattana and Dr Steven M. Smith, The University of Western Australia, for providing pGreen0049 mRFP-SRL and Ms Kimberley Tilbrook for providing pRFP-TS.
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