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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (05): 921-927.doi: 10.3724/SP.J.1006.2012.00921

• RESEARCH NOTES • Previous Articles     Next Articles

Optimization of Two-dimensional Electrophoresis Condition for Rice Grain Protein and Comparison of Relevant Proteomic Methods

LI Qi-Song,CHEN Jun,LIN Shi-Sheng,LI Zhong,ZHANG Zhi-Xing,LIN Wen-Xiong*   

  1. Institute of Agricultural Ecology, School of Life Sciences, Fujian Agriculture and Forestry University / Key Laboratory of Crop Physiology and Molecular Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2011-07-08 Revised:2011-12-19 Online:2012-05-12 Published:2012-03-05
  • Contact: 林文雄, E-mail: lwx@fjau.edu.cn E-mail:liqisong0591@163.com

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Abstract: A suitable proteomic strategy for rice grain protein research is important for further understanding rice grain development at proteome level. In this study, three different protein extraction methods were compared and then electrophoretic conditions of hand-made gel lands and IPG lands in IEF (isoelectric focusing) were optimized. Furthermore, three important proteomic research strategies (MALDI-TOF/MS, western-blot and phosphoproteome) were used for identification and analysis of in-gel proteins. The results showed that the method of soluble protein extraction was most suitable for investigation of grain proteomics, with a well-distributed 2-DE gel profile under optimized electrophoretic conditions, and it was confirmed that MALDI-TOF/MS, western-blot and phosphoproteome were the three suitable methods for the analysis of in-gel proteins. The establishment of the suitable system for rice grain proteomic research paves the way for the next step to further analyse rice grain development at proteome level.

Key words: Rice, Grain filling, Proteomics, Two-dimensional electrophoresis, Protein phosphorylation

[1]Teng Z-H(滕中华), Zhi L(智丽), Lü J(吕俊), Zong X-F(宗学凤), Wang S-G(王三根), He G-H(何光华). Effects of high temperature on photosynthesis characteristics, phytohormones and grain quality during filling-periods in rice. Acta Ecol Sin (生态学报), 2010, 30(23): 6504–6511 (in Chinese with English abstract)

[2]Yang J-C(杨建昌), Wang G-Z(王国忠), Wang Z-Q(王志琴), Liu L-J(刘立军), Zhu Q-S(朱庆森). Grain-filling characteristics and changes of hormonal content in the grains of dry-cultivated rice during grain-filling. Acta Agron Sin (作物学报), 2002, 28(5): 615–621 (in Chinese with English abstract)

[3]Wang H-Z(王贺正), Ma J(马均), Li X-Y(李旭毅), Zhang R-P(张荣萍). Effects of water stress on grain filling and activities of enzymes involved in starch synthesis in rice. Sci Agric Sin (中国农业科学), 2009, 42(5): 1550–1558 (in Chinese with English abstract)

[4]Zhu T, Budworth P, Chen W. Transcriptional control of nutrient partitioning during rice grain filling. Plant Biotechnol J, 2003, 1: 59–70

[5]Wan X Y, Liu J Y. Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves. Mol & Cell Proteomics, 2008, 7: 1469–1488

[6]Gallardo K, Job C, Groot S P C, Puype M, Demol H, Vandekerckhove J, Job D. Proteomics of Arabidopsis seed germination: a comparative study of wild-type and gibberellin-deficient seeds. Plant Physiol, 2002, 129: 823–837

[7]Houston N L, Hajduch M, Thelen J J. Quantitative proteomics of seed filling in castor: comparison with soybean and rapeseed reveals differences between photosynthetic and nonphotosynthetic seed metabolism. Plant Physiol, 2009, 151: 857–868

[8]Roccoa M, Corradob G, Arenac S, Ambrosioc C D, Tortiglioneb C, Sellarolid S, Marrad M, Raob R, Scalonic A. The expression of tomato prosystemin gene in tobacco plants highly affects host proteomic repertoire. J Proteomics, 2008, 71: 176–185

[9]Agrawal G K, Thelen J J. Large scale identification and quantitative profiling of phosphoproteins expressed during seed filling in oilseed rape. Mol Cell Proteomics, 2006, 5: 2044–2059

[10]Sheng B X, Tang L, Zhu Y D, Kang C, Yongbiao X, Tai W. Dynamic proteomic analysis reveals a switch between central carbon metabolism and alcoholic fermentation in rice filling grains. Plant Physiol, 2008, 148: 908–925

[11]Wang J-Y(王经源), Chen S-Y(陈舒奕), Liang Y-Y(梁义元), Lin W-X(林文雄). Improvement of ISO-DALT electrophoresis system. J Fujian Agric & For Univ (福建农林大学学报), 2006, 35(2): 187–190 (in Chinese with English abstract)

[12]Dumas-Gaudot E, Amiour N, Weidmann S, Bestel-Corre G, Valot S, Lenogue B, Gianinazzi-Pearsonl V, Gianinazzi S. A technical trick for studying proteomics in parallel to transcriptomics in symbiotic root–fungus interactions. Proteomics, 2004, 4: 451–453

[13]Laemmli U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 1970, 227: 680–685

[14]Blum H, Beiers H, Gross H J. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis, 1987, 8: 93–99

[15]Peng X X, Ye X T, Wang S Y. Identification of novel immunogenic proteins of Shigella flexneri 2a by proteomic methodologies. Vaccine, 2004, 22: 2750–2756

[16]Meng H(孟慧), Duan C-F(段翠芳), Zeng R-Z(曾日中). Researches of Plant Proteomics. China J Trop Agric (热带农业科学), 2006, 26(2): 60–64 (in Chinese with English abstract)

[17]Saravanan R S, Rose J K C. A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues. Proteomics, 2004, 4: 2522–2532

[18]Zhen Y(甄艳), Shi J-S(施季森). Application of mass spectrometry in proteomics studies. J Nanjing For Univ (南京林业大学学报), 2011, 35(1): 103–108 (in Chinese with English abstract)

[19]Cohen P. The origins of protein phosphorylation. Nat Cell Biol, 2002, 4: E127–E130
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