A bioinformatic approach to the identification of a conserved domain in a sugarcane legumain that directs GFP to the lytic vacuole
Mark A. Jackson A B C , Anne L. Rae A B D , Rosanne E. Casu A B , Christopher P. L. Grof A B , Graham D. Bonnett A B and Donald J. Maclean A CA Cooperative Research Centre for Sugar Industry Innovation through Biotechnology, University of Queensland, St Lucia, Qld 4072, Australia.
B CSIRO Plant Industry, Queensland Bioscience Precinct, 306 Carmody Rd, St Lucia, Qld 4067, Australia.
C School of Molecular and Microbial Sciences, University of Queensland, St Lucia, Qld 4072, Australia.
D Corresponding author. Email: anne.rae@csiro.au
Functional Plant Biology 34(7) 633-644 https://doi.org/10.1071/FP07024
Submitted: 2 February 2007 Accepted: 3 May 2007 Published: 4 July 2007
Abstract
Sugarcane is an ideal candidate as a biofactory for the production of alternate higher value products. One way of achieving this is to direct useful proteins into the vacuoles within the sugarcane storage parenchyma tissue. By bioinformatic analysis of gene sequences from putative sugarcane vacuolar proteins a motif has been identified that displays high conservation across plant legumain homologues that are known to function within vacuolar compartments. This five amino acid motif, represented by the sequence IRLPS in sugarcane is shown to direct an otherwise secreted GFP fusion protein into a large acidic and proteolytic vacuole in sugarcane callus cells as well as in diverse plant species. In mature sugarcane transgenic plants, the stability of GFP appeared to be dependent on cell type, suggesting that the vacuolar environment can be hostile to introduced proteins. This targeting motif will be a valuable tool for engineering plants such as sugarcane for production of novel products.
Additional keywords: biofactory, Saccharum, vacuole processing enzyme, vacuole targeting.
Acknowledgements
This research was undertaken with PhD research scholarship funding from the Cooperative Research Centre for Sugar Industry Innovation through Biotechnology. The nucleotide sequence of the sugarcane legumain-like gene has been submitted to GenBank under the accession number DQ458784. The authors wish to thank Dr Frank Smith and Dr Peer Schenk for kindly providing vectors used in this study. The authors also wish to thank Dr Gang Ping Xue, Dr Kerry Nutt, Dr Jason Geijskes and Jai Perroux for technical advice and assistance.
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