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

Journal of Experimental Botany
Robert ParkerNicholas V J Harpham

Abstract

Proteomic analysis of any biological system by two-dimensional 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 93-95% 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 long-term 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 pro...Continue Reading

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Citations

Feb 12, 2008·Amino Acids·K AghaeiS Komatsu
Jul 5, 2006·Functional & Integrative Genomics·Chandan SahiAnil Grover
Jan 10, 2009·Plant & Cell Physiology·Yuichi Tada, Takaaki Kashimura
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Feb 14, 2014·Plant Physiology and Biochemistry : PPB·Shahniyar Bayramov, Novruz Guliyev
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Dec 15, 2015·Journal of Proteomics·Abu Hena Mostafa Kamal, Setsuko Komatsu
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Aug 8, 2012·Plant Physiology and Biochemistry : PPB·Elham SarhadiGhasem Hosseini Salekdeh
Jan 13, 2012·Journal of Proteomics·Yu-Dong WangYum-shing Wong
Jul 4, 2009·Peptides·Roya RazavizadehSetsuko Komatsu
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Mar 21, 2013·Proteomics·Bronwyn J BarklaOmar Pantoja
Apr 12, 2011·Proteomics·Ali BandehaghSetsuko Komatsu

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