1. 中国农业大学资源与环境学院, 农业部植物营养学重点开放实验室，北京 100193 2. Center for Plant Molecular Biology, Department of General Genetics, Eberhard Karls University Tuebingen, 72076 Tuebingen, Germany
Comparative Studies on Identification of Maize Secreted Proteome by Different Analyzing Techniques
MA Wei1, Frank Hochholdinger2, LI Chun-jian1*
1. The Key Laboratory of Plant Nutrition, Ministry of Agriculture，Department of Plant Nutrition, China Agricultural University, Beijing 100193, China 2. Center for Plant Molecular Biology, Department of General Genetics, Eberhard Karls University Tuebingen, 72076 Tuebingen, Germany
Abstract：Maize roots release a battery of proteins into the rhizosphere during growth and development. In the present study, mucilage secreted by primary roots of 3-4 d old maize seedlings was collected under axenic conditions, and the constitutively secreted proteins were analyzed by different methods, including two-dimensional electrophoresis -MADLI-TOF, LC-MS, and Shotgun techniques. The results showed that 200 protein spots could be observed on two-dimensional electrophoresis gel stained by silver nitrate. However, due to the low protein content for each protein spot, there was not reliable result after identification by MADLI-TOF. On the other hand, 152 and 2 848 proteins were identified by LC/MS and Shotgun technique, respectively. All 152 proteins identified by LC/MS method could be found in the proteins identified by Shotgun method, and thus the results obtained by LC/MS and Shotgun method could be authenticated mutually. The results demonstrated that the Shotgun method is more sensitive than the other methods for protein identification, especially for the proteins with lower contents and complex interfering substances, such as the plant root released proteins.
马 玮1, Frank Hochholdinger2, 李春俭1* . 用不同蛋白质分析方法鉴定玉米分泌蛋白组的比较研究 [J]. 光谱学与光谱分析, 2010, 30(10): 2762-2766.
MA Wei1, Frank Hochholdinger2, LI Chun-jian1* . Comparative Studies on Identification of Maize Secreted Proteome by Different Analyzing Techniques . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2010, 30(10): 2762-2766.
[1] ZHANG Fu-suo, CAO Yi-ping(张福锁，曹一平). Acta Pedol. Sin.(土壤学报)，1992, 29(3): 239. [2] ZHANG Fu-suo (张福锁). J. China Agricultural University(北京农业大学学报), 1992, 18(4): 353. [3] Marschner H. Mineral Nutrition of Higher Plants. 2nd. London: Academic Press, 1995. [4] SHEN Jian-bo, ZHANG Fu-suo(申建波，张福锁). Journal of China Agric. Sci. Tech.,(中国农业科技导报), 1999, 1(4): 21. [5] Bais H P, Park S W, Weir T L, et al. Trends Plant Sci., 2004, 9(1): 26. [6] ZHANG Nan-nan, LIU Xin, SUN Jing, et al(张楠楠，刘 欣，孙 晶，等). Hereditas(遗传), 2009, 31(1): 29. [7] Chevallet M, Diemer H, Van Dorssealer A, et al. Proteomics, 2007, 7(11): 1757. [8] Basu U, Francis J L, Whittal R M, et al. Plant Soil, 2006, 286(1): 357. [9] Wen F S, Van Etten H D, Tsaprailis G, et al. Plant Physiol., 2007, 143(2): 773. [10] ZHANG Yang-jun, CAI Yun, WANG Jing-lan, et al(张养军，蔡 耘，王京兰，等). Chinese Journal of Chromatography(色谱), 2003, 21(1): 20. [11] McDonald W H, Yates J R. Curr. Opin. Mol. Ther., 2003, 5(3): 302. [12] Blum W F, Ranke M B, Lechner B, et al. Acta Endocrinol (Copenh), 1987, 116(4): 445. [13] Rappsilber J, Ishihama Y, Mann M. Anal. Chem., 2003, 75: 663. [14] Rappsilber J, Mann M, Ishihama Y. Nat. Protoc., 2007, 2: 1896. [15] Ma W, Muthreich N, Liao C S, et al. J. Proteome Res., 2010, 9: 2968. [16] Wolters D A, Washburn M P, Yates J R. Anal. Chem., 2001, 73(23): 5683. [17] Hooler T F, Jorgenson J W. Anal. Chem., 1997, 69(20): 4134. [18] Link A J, Eng J, Schieltz D M, et al. Nature Biotechnol., 1999, 17(7): 676. [19] Koller A, Washburn M, Plange B M, et al. Pro. National Academy of Sciences, 2002, 99(18): 1969. [20] Schirmer E C, Florens L, Guan T L, et al. Science, 2003, 301: 1380. [21] Peng J M，Schwartz D，Elias J E, et al. Nature Biotechnol., 2003, 21(8): 921. [22] Ideker T, Thorsson V, Ranish J A, et al. Sciences, 2001, 292: 929. [23] Lasonder E, Ishihama Y, Andersen J S, et al. Nature, 2002, 419: 537. [24] Andersen J S, Wilkinson C J, Mayort T, et al. Nature, 2003, 426: 570. [25] Fischer F, Wolters D, Rogner M, et al. Mol. Cell. Proteomics, 2006, 5(3): 444.
