Abstract
Two-dimensional difference gel electrophoresis is an invaluable technique for the analysis of plant proteomes. However, preparation of protein fractions from plant tissues is challenging due to the special features of plant cells: a robust cell wall, large vacuoles which often contain high concentrations of organic acids and a broad range of secondary metabolites like phenolic compounds and pigments. Therefore, protein preparation for difference gel electrophoresis (DIGE) analyses has to be adapted. Here, we describe both a phenolic protein extraction method for plant tissues and an adapted protocol for DIGE labeling of the generated fractions.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Raggiaschi R., Lorenzetto C., Diodato E., Caricasole A., Gotta S. and Terstappen G.C. (2006) Detection of phosphorylation patterns in rat cortical neurons by combining phosphatase treatment and DIGE technology. Proteomics 6, 748–756.
Lyngved R., Renaut J., Hausman J.F., Iversen T.H., and Hvoslef-Eide A.K. (2008) Embryo-specific Proteins in Cyclamen persicum analyzed with 2-D DIGE. J Plant Growth Regul 27, 353–369.
Gomez A., Lopez J.A., Pintos B., Camafeita E., and Bueno M.A. (2010) Proteomic analysis from haploid and diploid embryos of Quercus suber L. identifies qualitative and quantitative differential expression patterns. Proteomics 9, 4355–4367.
Rode C., Braun H.P., Gallien S., Heintz D., Van Dorsselaer A., and Winkelmann T. (2010) Establishment of proteome reference maps for somatic and zygotic embryos of Cyclamen persicum Mill. Acta Hort 855, 239–242.
Lilley K.S., and Dupree P. (2006) Methods of quantitative proteomics and their application to plant organelle characterization. J Exp Bot 57, 1493–1499.
Heinemeyer J., Scheibe B., Schmitz U.K., and Braun H.P. (2009) Blue native DIGE as a tool for comparative analyses of protein complexes. J Prot 72, 539–544.
Ndimba B.K., Chivasa S., Simon W.J., and Slabas A.R. (2005) Identification of Arabidopsis salt and osmotic stress responsive proteins using two-dimensional difference gel electrophoresis and mass spectrometry. Proteomics 5, 4185–4196.
Kieffer P., Schroder P., Dommes J., Hoffmann L., Renaut J., and Hausman J.F. (2009) Proteomic and enzymatic response of poplar to cadmium stress. J Proteom 72, 379–396.
Chivasa S., Hamilton J.M., Pringle R.S., Ndimba B.K., Simon W.J., Lindsey K., and Slabas A.R. (2006) Proteomic analysis of differentially expressed proteins in fungal elicitor-treated Arabidopsis cell cultures. J Exp Bot 57, 1553–1562.
Van Noorden G.E., Kerim T., Goffard N., Wiblin R., Pellerone F.I., Rolfe B.G., and Mathesius U. (2007) Overlap of proteome changes in Medicago truncatula in response to auxin and Sinorhizobium meliloti. Plant Physiol 144, 1115–1131.
Amey R.C., Schleicher T., Slinn J., Lewis M., Macdonald H., Neill S.J., and Spencer-Phillips PTN. (2008) Proteomic analysis of a compatible interaction between Pisum sativum (pea) and the downy mildew pathogen Peronospora viciae. Eur J Plant Pathol 122, 41–55.
Schenkluhn L., Hohnjec N., Niehaus K., Schmitz U.K., and Colditz F. (2010) Differential gel electrophoresis (DIGE) to quantitatively monitor early symbiosis- and pathogenesis-induced changes of the Medicago truncatula root proteome. J Proteom 73, 753–768.
Dornez E., Croes E., Gebruers K., Carpentier S., Swennen R., Laukens K., Witters E., Urban M., Delcour J.A., and Courtin C.M. (2010) 2-D DIGE reveals changes in wheat xylanase inhibitor protein families due to Fusarium graminearum Delta Tri5 infection and grain development. Proteomics 10, 2303–2319.
Tang W.Q., Deng Z.P., Oses-Prieto J.A., Suzuki N., Zhu S.W., Zhang X., Burlingame A.L., and Wang Z.Y. (2008) Proteomics studies of brassinosteroid signal transduction using prefractionation and two-dimensional DIGE. Mol Cell Proteom 7, 728–738.
Gallardo K., Job C., Groot S.P.C., Puype M., Demol H., Vandekerckhove J., and Job, D. (2002) Proteomics of Arabidopsis seed germination: a comparative study of wildtype and gibberelline deficient seeds. Plant Physiol 129, 823–837.
Damerval C., De Vienne D., Zivy M., and Thiellement H. (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins. Electrophoresis 7, 52–54.
Corke F.M.K., and Roberts K. (1996) Large changes in the population of cell wall proteins accompany the shift to cell elongation. J Exp Bot 48, 971–977.
Laemmli U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Hurkman W.J., and Tanaka C.K. (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol 81, 802–806.
Colditz F., Nyamsuren O., Niehaus K., Eubel H., Braun H.-P., and Krajinski F. (2004) Proteomic approach: Identification of Medicago truncatula proteins induced in roots after infection with the pathogenic oomycete Aphanomyces euteiches. Plant Mol Biol 55, 109–120.
Schägger H., and von Jagow G. (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166, 368–379.
Führs H., Hartwig M., Buitrago Molina L.E., Heintz D., Van Dorsselaer A., Braun H.-P., and Horst W.J. (2008) Early manganese-toxicity response in Vigna unguiculata L. – a proteomic and transcriptomic study. Proteomics 8, 149–159.
Acknowledgments
The authors would like to thank Jennifer Klodmann, Institute for Plant Genetics, Leibniz Universität Hannover, for critically reading the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Rode, C., Winkelmann, T., Braun, HP., Colditz, F. (2012). DIGE Analysis of Plant Tissue Proteomes Using a Phenolic Protein Extraction Method. In: Cramer, R., Westermeier, R. (eds) Difference Gel Electrophoresis (DIGE). Methods in Molecular Biology, vol 854. Humana Press. https://doi.org/10.1007/978-1-61779-573-2_23
Download citation
DOI: https://doi.org/10.1007/978-1-61779-573-2_23
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-572-5
Online ISBN: 978-1-61779-573-2
eBook Packages: Springer Protocols