通过无公害化学诱导子的诱导可使茶树芽叶的EGCG含量提高20.15%~25.00%。为了探明诱导的分子机制,用固相pH梯度双向凝胶电泳分离诱导芽叶与正常芽叶的总蛋白质,结果获得了分辨率和重复性较好的双向电泳图谱,差异表达分析发现,诱导出现的特异蛋白有14种,诱导消失的特异蛋白有8种,诱导表达上调相差10倍的特异蛋白有11种, 诱导表达下调相差10倍的特异蛋白有6种。选取两个差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)测定其酶解后的肽质指纹图谱,通过http:/www.matrixscience.com网站,利用Mascot软件检索NCBInr数据库。查询结果:一种为光合系统Ⅰ的铁硫蛋白,另一种为未知蛋白。这些结果表明,茶树诱导芽叶与正常芽叶的蛋白质组存在差异,这些特异蛋白可能在诱导过程中起着重要的作用。
LIN Jin-ke
,
ZHENG Jin-gui
,
YUAN Ming
,
ZHANG Xue-qin
,
Wang Feng
. 外源诱导提高茶树EGCG含量过程的蛋白质组差异分析[J]. 茶叶科学, 2005
, 25(2)
: 109
-115
.
DOI: 10.13305/j.cnki.jts.2005.02.005
Induced by harmless inducer, the EGCG content of tea shoots improved 20.15%~25.00%. To probe into the molecular mechanism, the immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis was used to separate the total proteins of induced tea shoots and normal tea shoots. The two- dimensional polyacrylamide gel electrophoresis map with high resolution and repeatability was produced. Differential expression analysis indicated that 14 specific proteins were emerged in induced shoot and 8 disappeared, 11 expressed above ten-fold, and 6 below one-tenth. Two differential protein spots were selected for MALDI-TOF-MS to determine the post-enzymolysis peptide mass fingerprinting. Using Mascot to search NCBInr database on website at http://www.matrixscience.com. The results were as follows: one was photosystem I iron-sulfur protein, and the another unkown. The results showed that there were proteome difference between induced and normal tea shoots, that the specific protein might play an important part during inducing process.
[1] Jankun J, Selman S H, Swiercz R. Why drinking green tea could prevent cancer[J]. Nature, 1997, 387(6633): 561~564.
[2] Lin Jin-ke, Zheng Jin-gui. Effects on the quality biochemical components of fresh leaves in the process of inducing and improving the content of EGCG in tea[J]. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2004, 33(1): 65~68(in Chinese).
[3] Zhu Yong-xing. Study and using of tea plant growth regulating mater. China Tea, 1999(3): 12~13.
[4] Joachim Klose, Christina N, Marion Herrmann Genetic analysis of the mouse brain proteome[J]. Nature, 2002, 01: 385 ~393.
[5] Ott V, Guenther K, Steinert R, et al. Accuracy of two-dimensional electrophoresis for target discovery in human colorectal cancer s[J]. Nature, 2001 Aug 10, 142~151.
[6] Chen Rong-zhi, Weng Qing-mei, Huang Zhen, et al. Analysis of Resistance-related Proteins in Rice Against Brown Planthopper by Two-dimensional Electrophoresis[J]. Acta Botanica Sinica, 2002, 44(4): 427~432.
[7] Yu Li-rong, Wang Nan, Wu Gao-de. Two-dimensional Polyacrylamide Gel Electrophoresis Analysis of HumanLiver Cell BEL-7404 Cell and of Normal Cell L-02[J]. Chinese Science Bulletin, 2000, 45(2): 170~178(in Chinese).
[8] Cheng Hai-pin, Qian Xiao-hong. The Technological System of Proteme Research and Its Progress[J]. Prog Biochem Biophys, 2000, 27(6): 584~588(in Chinese).
[9] Wu Qing-zhi, Huang Kai-xun, Xun Hui-bi. Progress in Two-dimensional Electrophoresis Technique[J]. Letters in Biotechnology, 2002, 13(2): 28~311(in Chinese).
[10] Yang He-yi Ying Wang-tao, Qian Xiao-hong. Research Progress of Proteome[J]. Technology Progress of Science, 2002, 12(1): 13~19(in Chinese).
[11] Dung Hai-feng, Qian Ling-jia, Ying Wang-tao, et al. Two-dimensional Polyacrylamide Gel Electrophoresis Analysis of Stressed Cardiomyocytes Proteome[J]. Chinese Juornal of Biochemistry and Molecular Biology. 2002, 18(2): 234~239(in Chinese).
[12] Yu Bin, Lin Bing. One Improvement Method of Isolation and Two-Dimensional Electrophoresis of plant Protein[J]. Prog.Biochem.Biophys, 1989, 16(6): 480~481 (in Chinese).
[13] Wan Xiao-chun. Tea Biochemistry[M]. Beijing: China Agricultural Press, 2003, 9~20.
[14] Lin Jin-ke, Zheng Jin-gui Yuan Ming, et al. The Improvement of Isolation and The Method of Two-Dimensional Electrophoresis of Protein from Tea Plant[J]. Journal of Tea Science, 2003, 21(1): 16~20(in Chinese).
[15] Guo yao-jun. Protein Electrophoresis Technique[M]. Beijing: Science Press, 2001, 141~144.
[16] Tong Qi-qin. Tea Plant Cultivation[M]. Beijing: China Agricultural Press, 2000, 92~94.