应用蛋白质组学的方法研究拟南芥根系响应环境因子做出的蛋白质差异表达时，需要提取根系的全部信息。过去拟南芥的培养方法不能满足蛋白质组学研究对根系样本在质和量两个方面的严格需求，个别能够解决这个问题的方案又因为价格昂贵难以推广。本文在前人研究的基础上开发了一种简单高效的拟南芥无菌水培系统。随机抽取这个系统培养的拟南芥根系分别进行双向电泳实验，通过蛋白质点匹配，等点聚焦和SDS-PAGE方向坐标偏差分析，以及相对含量变异系数分析，证明这个水培系统可以满足蛋白质组学对实验材料苛刻的要求。
Abstract
When apply proteomics approaches to investigate Arabidopsis roots' responses to environment factors, gaining all information form roots are always required. Due to major technical obstacle, previous culture methods were hard to meet the strict standards of proteomics analysis. Based on past methods, an easy, low-cost and contamination-free hydroponic cultivation system for Arabidopsis is introduced. Random seedlings in this system were selected to perform two-dimensional electrophoresis. The results of spots detecting, position deviation in the isoelectric focusing and SDS-PAGE, and average coefficient of variability of spot volume were proved this system is excellent for Arabidopsis proteomics analysis.
关键词
拟南芥; 无菌水培; 根系; 蛋白质组学
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Key words
Arabidopsis thaliana; hydroponic culture under sterile conditions; roots; proteomic
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参考文献
参考文献
[1]Sacha B, Wilhelm G. Arabidopsis thaliana proteomics: from proteome to genome[J]. J Exp Bot, 2006,57(7):1485-1491
[2]Mehta A, Magalhães BS, Souza DS et al. Rooteomics: the challenge of discovering plant defense-related proteins in roots[J]. Curr Pro Pep Sci, 2008, 9(2):108-116
[3]郁晓敏,方萍,朱日清.拟南芥直播水培法[J].植物生理学通讯,2004,40(1):81-82
[4]Gibeaut DM, Hulett J, Cramer GR et al. Maximal biomass of Arabidopsis thaliana using a simple, low maintenance hydroponic method and favorable environmental conditions[J]. Plant Physiol, 1997,115(2):317-319
[5]Huttner D, Bar-Zvi D. An improved, simple, hydroponic method for growing Arabidopsis thaliana[J]. Plant Mol Biol Rep, 2003, 21(1): 59-63
[6]Karen S, Joske R, Frank VB et al. Critical evaluation and statistical validation of a hydroponic culture system for Arabidopsis thaliana[J]. Plant Physiol Biochem, 2008,46(2):212-218.
[7]赵淑清, 郭剑波. 一种简单快速的拟南芥水培方法的研究[J].生物学杂志, 2000,17(6): 22-23.
[8]Arteca RN, Arteca JM. A novel method for growing Arabidopsis thaliana plants hydroponically[J]. Physiol Plant, 2000, 108(2): 188-193
[9]Siedlecka A, Krupa Z. Simple method of Arabidopsis thaliana cul-tivation in liquid nutrient medium[J]. Acta Physiol Plant, 2002, 24(2): 163-166
[10]陈建红,沈宏.一种拟南芥无菌直播水培技术[J].植物生理学通讯, 2007, 43(2): 348
[11]Bernhard S, Feédéric B, Hans-Peter M. A hydroponic culture system for growing Arabidopsis thaliana plantlets under sterile conditions[J]. Plant Mol Biol Rep, 2003, 21(4): 449-456
[12]Tal I, Cynthia MB, Ramu SS et al. Sample extraction techniques for enhanced proteomic analysis of plant tissues[J]. Nature Protocols, 2006,1(1):769-774
[13]Neuhoff V, Arold N, Taube D et al. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250[J]. Electrophoresis, 1988, 9(6): 255-262.
[14]Alonso-Blanco C, Blankestijn-De Vries H, Hanhart CJ et al. Natural allelic variation at seed size loci in relation to other life history traits of Arabidopsis thaliana[J]. Proc Natl Acad Sci, 1999,96(8):4710-4717.
[15]Julio S, Jose JS. Methods in Molecular Biology, vol. 323: Arabidopsis Protocols[M], 2nd Edition, Humana Press 2006.
[16]Tocquin P, Corbesier L, Havelange A et al. A novel high efficiency, low maintenance, hydroponic system for synchronous growth and flowering of Arabidopsis thaliana[J].BMC Plant Biology, 2003,(3):2.(http://www.biomedcentral.com/1471-2229/3/2)
[17]俞利荣,王楠,吴高德,等.人肝癌细胞系BEL-7404和正常肝细胞系L-02表达的蛋白质组双向凝胶电泳分析[J].科学通报,2000,45(2):170-178.
[18]Joseph MC, Michael JD, Anton P et al. Positional reproducibility of protein spots in two-dimensional polyacrylamide gel electrophoresis using immobilised pH gradient isoelectric focusing in the first dimension-An interlaboratory comparison[J]. Electrophoresis, 1994,15(1):1205-1211.
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