Welcome to Acta Agronomica Sinica,

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

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
[1] TIAN Tian, CHEN Li-Juan, HE Hua-Qin. Identification of rice blast resistance candidate genes based on integrating Meta-QTL and RNA-seq analysis [J]. Acta Agronomica Sinica, 2022, 48(6): 1372-1388.
[2] ZHENG Chong-Ke, ZHOU Guan-Hua, NIU Shu-Lin, HE Ya-Nan, SUN wei, XIE Xian-Zhi. Phenotypic characterization and gene mapping of an early senescence leaf H5(esl-H5) mutant in rice (Oryza sativa L.) [J]. Acta Agronomica Sinica, 2022, 48(6): 1389-1400.
[3] ZHOU Wen-Qi, QIANG Xiao-Xia, WANG Sen, JIANG Jing-Wen, WEI Wan-Rong. Mechanism of drought and salt tolerance of OsLPL2/PIR gene in rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1401-1415.
[4] ZHENG Xiao-Long, ZHOU Jing-Qing, BAI Yang, SHAO Ya-Fang, ZHANG Lin-Ping, HU Pei-Song, WEI Xiang-Jin. Difference and molecular mechanism of soluble sugar metabolism and quality of different rice panicle in japonica rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1425-1436.
[5] YAN Jia-Qian, GU Yi-Biao, XUE Zhang-Yi, ZHOU Tian-Yang, GE Qian-Qian, ZHANG Hao, LIU Li-Jun, WANG Zhi-Qin, GU Jun-Fei, YANG Jian-Chang, ZHOU Zhen-Ling, XU Da-Yong. Different responses of rice cultivars to salt stress and the underlying mechanisms [J]. Acta Agronomica Sinica, 2022, 48(6): 1463-1475.
[6] CHEN Jing, REN Bai-Zhao, ZHAO Bin, LIU Peng, ZHANG Ji-Wang. Regulation of leaf-spraying glycine betaine on yield formation and antioxidation of summer maize sowed in different dates [J]. Acta Agronomica Sinica, 2022, 48(6): 1502-1515.
[7] XU Tian-Jun, ZHANG Yong, ZHAO Jiu-Ran, WANG Rong-Huan, LYU Tian-Fang, LIU Yue-E, CAI Wan-Tao, LIU Hong-Wei, CHEN Chuan-Yong, WANG Yuan-Dong. Canopy structure, photosynthesis, grain filling, and dehydration characteristics of maize varieties suitable for grain mechanical harvesting [J]. Acta Agronomica Sinica, 2022, 48(6): 1526-1536.
[8] YANG Jian-Chang, LI Chao-Qing, JIANG Yi. Contents and compositions of amino acids in rice grains and their regulation: a review [J]. Acta Agronomica Sinica, 2022, 48(5): 1037-1050.
[9] DENG Zhao, JIANG Nan, FU Chen-Jian, YAN Tian-Zhe, FU Xing-Xue, HU Xiao-Chun, QIN Peng, LIU Shan-Shan, WANG Kai, YANG Yuan-Zhu. Analysis of blast resistance genes in Longliangyou and Jingliangyou hybrid rice varieties [J]. Acta Agronomica Sinica, 2022, 48(5): 1071-1080.
[10] YANG De-Wei, WANG Xun, ZHENG Xing-Xing, XIANG Xin-Quan, CUI Hai-Tao, LI Sheng-Ping, TANG Ding-Zhong. Functional studies of rice blast resistance related gene OsSAMS1 [J]. Acta Agronomica Sinica, 2022, 48(5): 1119-1128.
[11] ZHU Zheng, WANG Tian-Xing-Zi, CHEN Yue, LIU Yu-Qing, YAN Gao-Wei, XU Shan, MA Jin-Jiao, DOU Shi-Juan, LI Li-Yun, LIU Guo-Zhen. Rice transcription factor WRKY68 plays a positive role in Xa21-mediated resistance to Xanthomonas oryzae pv. oryzae [J]. Acta Agronomica Sinica, 2022, 48(5): 1129-1140.
[12] WANG Xiao-Lei, LI Wei-Xing, OU-YANG Lin-Juan, XU Jie, CHEN Xiao-Rong, BIAN Jian-Min, HU Li-Fang, PENG Xiao-Song, HE Xiao-Peng, FU Jun-Ru, ZHOU Da-Hu, HE Hao-Hua, SUN Xiao-Tang, ZHU Chang-Lan. QTL mapping for plant architecture in rice based on chromosome segment substitution lines [J]. Acta Agronomica Sinica, 2022, 48(5): 1141-1151.
[13] WANG Ze, ZHOU Qin-Yang, LIU Cong, MU Yue, GUO Wei, DING Yan-Feng, NINOMIYA Seishi. Estimation and evaluation of paddy rice canopy characteristics based on images from UAV and ground camera [J]. Acta Agronomica Sinica, 2022, 48(5): 1248-1261.
[14] KE Jian, CHEN Ting-Ting, WU Zhou, ZHU Tie-Zhong, SUN Jie, HE Hai-Bing, YOU Cui-Cui, ZHU De-Quan, WU Li-Quan. Suitable varieties and high-yielding population characteristics of late season rice in the northern margin area of double-cropping rice along the Yangtze River [J]. Acta Agronomica Sinica, 2022, 48(4): 1005-1016.
[15] CHEN Yue, SUN Ming-Zhe, JIA Bo-Wei, LENG Yue, SUN Xiao-Li. Research progress regarding the function and mechanism of rice AP2/ERF transcription factor in stress response [J]. Acta Agronomica Sinica, 2022, 48(4): 781-790.
Full text



No Suggested Reading articles found!