An accurate and reproducible method for proteome profiling of the effects of salt stress in the rice leaf lamina

Parker, Robert, Flowers, Timothy J, Moore, Anthony L and Harpham, Nicholas V J (2006) An accurate and reproducible method for proteome profiling of the effects of salt stress in the rice leaf lamina. Journal of Experimental Botany, 57 (5). pp. 1109-1118. ISSN 0022-0957

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Abstract

Proteomic analysis of any biological system by twodimensional gel electrophoresis (2-DE) requires high resolution and high reproducibility. The results presented here demonstrate the reproducible and accurate separation of rice (Oryza sativa L.) proteins using improved procedures for high resolution 2-DE, which were adapted for the separation of rice lamina proteins. Validation of this system was achieved by measuring the effects of sample preparation and biological variation on the coefficient of variation (CV) for replicate spots. The majority of experimental variation was shown to be introduced by the 2-DE technique (CV 0.26). Analysis of biological variation indicated that approximately 9395% of spots were within a CV of 0.7. This provided a threshold value from which valid differences in expression between experimental groups could be screened. This system was then utilized for the proteomic analysis of short- and longterm salt-stress-responsive proteins in the rice leaf lamina. Analysis resulted in the separation of approximately 2500 protein species of which 32 were observed to be significantly regulated by salinity; so far 11 of these proteins have been identified by tandem mass spectrometry. An increase in eight proteins, including RuBisCO activase and ferritin, occurred by 24 h of exposure to sodium chloride (50 mM) and continued to increase during the following 6 d. Only one protein, a putative phosphoglycerate kinase, was found to increase in expression within 24 h and did not increase over a longer period of exposure to salt. There were also proteins that showed no change 24 h after exposure to salt, but had increased (superoxide dismutase) or decreased (S-adenosyl-L-methionine synthetase) after 7 d salt treatment.

Item Type: Article
Additional Information: A rigorous evaluation of methods for analysis of the effects of salinity on the proteome of rice. Parker was a DPhil student, the biological side of whose research was directed by Flowers: this continued a long sequence of his research on salt-tolerance of rice. Harpham provided technical expertise on proteomics.
Schools and Departments: School of Life Sciences > Evolution, Behaviour and Environment
Subjects: Q Science > QK Botany > QK0710 Plant physiology
Depositing User: Tim Flowers
Date Deposited: 06 Feb 2012 20:33
Last Modified: 15 May 2012 08:34
URI: http://sro.sussex.ac.uk/id/eprint/26620
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