Optimization of EDTA Exudation Technique for Proteome Study of the Phloem
DOI:
https://doi.org/10.11113/jt.v64.2033Keywords:
EDTA exudation, phloem, proteomics, Arabidopsis, 1D-PAGE, 2D-PAGEAbstract
Phloem proteome study is gaining recognition due to the phloem important function to a plant. Identification of proteins within the phloem with varying function could help understand the regulation of growth and development. EDTA exudation technique applied to obtain Arabidopsis phloem protein was optimized. It was found that plants grown up until 6 weeks in SD (short day) condition, exposed to 3 days LD (long day) condition, the phloem sap collected in 20 mM EDTA with overnight incubation in LD and precipitated using Amicon filter device gave the optimum amount of protein with good quality. Sufficient amount of protein was obtained using the optimized EDTA exudation technique for protein identification.
References
Aki, T., Shigyo, M., Nakano, R., Yoneyama, T., Shuichi, Y. 2008. Nano Scale Proteomics Revealed The Presence of Regulatory Proteins Including Three FT-Like Proteins In Phloem And Xylem Saps From Rice. Plant Cell Physiol. 49(5): 767–790.
Batailler, B., Lemaitre, T., Vilaine, F., Sanchez, C., Renard, D., Cayla, T., Beneteau, J., Dinant, S. 2012. Soluble and Filamentous Proteins in Arabidopsis Sieve Elements. Plant, Cell and Environment. 35(7): 1258–1273.
Chabarek, S., Martell., A. E. 1959. Organic Sequestering Agents. John Wiley.
Corbesier, L., Vincent, C., Jang, S., Fornara, F., Fan, Q., Searle, I., Giakountis, A., Farrona, S., Gissot, L., Turnbull, C., Coupland, G. 2007. FT Protein Movement Contributes to Long-Distance Signalling in Floral Induction of Arabidopsis. Science. 316(5827): 1030–1033.
Giavalisco, P., Kapitza, K., Kolasa, A., Buhtz, A., Kehr, J. 2006. Towards The Proteome of Brassica napus Phloem Sap. Proteomics 6(3): 896–909.
Guelette, B. S., Benning, U. F., Hoffmann-Benning, S. 2012. Identification of Lipids and Lipid-binding Proteins in Phloem Exudates from Arabidopsis Thaliana. J Exp. Bot. 63(10): 3603–3616.
Hoffmann-Benning, S., Gage, D. A., McIntosh, L., Kende, H. and Zeevaart, J. A. D. 2002. Comparison of Peptides in the Phloem Sap of Flowering and Non-Flowering Perilla and Lupine Plants using Microbore HPLC Followed by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. Planta. 216: 140–147.
Lin, M-K., Lee, Y-J., Lough, T. J., Phinney, B. S. & Lucas, W. J. 2009. Analysis of the Pumpkin Phloem Proteome Provides Insights into Angiosperm Sieve Tube Function. Molecular & Cellular Proteomics. 8(2): 343–356.
King, R. W., Zeevart, A. D. 1974. Enhancement of Phloem Exudation from Cut Petioles by Chelating Agents. Plant Physiol. 53: 96–103.
Knoblauch, M., Peters, W. S., Ehlers, K., van Bel, A. J. E. 2001. Reversible Calcium-Regulated Stopcocks in Legume Sieve Tubes. Plant Cell. 13: 1221–1230.
Ponder, K. L., Pritchard, J., Harrington, R., Bale, J. S. 2000. Difficulties in Location and Acceptance of Phloem Sap Combined with Reduced Concentration of Phloem Amino Acids Explained Lowered Performance of the Aphid Rhopalosiphum padi on Nitrogen Deficient Barley (Hordeum vulgare) Seedlings. Entomologia Experimentalis et Applicata. 97: 203–210.
Tully, R. E., Hanson, A. D. 1979. Amino Acids Translocated From Turgid and Water-Stressed Barley Leaves. I. Phloem Exudation Studies. Plant Physiol. 64: 460–466.
Wilkinson, T. L., Douglas, A. E. 2003. Phloem Amino Acids and the Host Plant Range of the Polyphagous Aphid, Aphis fabae. Entomologia Experimentalis et Applicata. 106: 103–113.
Will, T., van Bel, A. J. E. 2006. Physical and Chemical Interactions Between Aphids and Plants. J Exp. Bot. 57: 729–737.
Yoshida, H., Tsumuki, H., Kawada, K., Kanehisa, K. 1996. Gramine and Resistance of Barley to Aphids: Analysis of EDTA Exudates from Barley Leaves. Bull. Res. Inst. Bioresour. Okayama Univ. 4: 73–78.
Downloads
Published
Issue
Section
License
Copyright of articles that appear in Jurnal Teknologi belongs exclusively to Penerbit Universiti Teknologi Malaysia (Penerbit UTM Press). This copyright covers the rights to reproduce the article, including reprints, electronic reproductions, or any other reproductions of similar nature.
