中国3个主栽烟草品种的差异蛋白质组学研究

doi:10.3864/j.issn.0578-1752.2016.16.003

Abstract

Abstract: 【Objective】 In order to strengthen the basic scientific research and cultivation of tobacco, differential proteomics is used to select specific proteins and elucidate metabolic pathways in tobacco. 【Method】 Leaf proteins of three varieties of tobacco in China including Honghuadajinyuan, K326 and Yunyan 87 were extracted by phenol. After digestion the protein profiles of three varieties of tobacco were investigated by using tandem mass tag(TMT) coupled with two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS). The proteins and peptides from tobacco were identified with MASCOT search engine after spectra extraction in Proteome Discoverer Software. Then, bioinformatics analysis including data correlation analysis, hierarchical cluster analysis and principal component analysis (PCA) were conducted. Meanwhile, proteins with change ratio of more than 2 fold were defined as differentially expressed tobacco proteins, and they were screened out by volcano plot analysis. Finally, the distribution and function of important proteins in tobacco were analyzed using KEGG pathway analysis. 【Result】 A total of 3 079 proteins in Honghuadajinyuan, K326 and Yunyan 87 were identified and the number of protein ID was 10 343. Protein expression profiles of hierarchical cluster analysis and principal component analysis indicated that K326 and Yunyan 87 were relatively similar, and Honghuadajinyuan was significantly different from the other two. The subsequent protein screening also verified the results. Screening of differentially expressed proteins suggested that there were only 29 proteins differentially expressed between K326 and Yunyan 87. 13 proteins of them covered 8 metabolic pathways including flavonoids biosynthesis. Chalcone synthetase participating in the flavonoids synthesis was significantly higher in K326 than in Yunyan 87. Honghuadajinyuan and Yunyan 87 had 160 differentially expressed proteins. 103 proteins of them covered 42 metabolic pathways including glutathione metabolism. Three kinds of enzymes related to glutathione metabolic pathway including glutathione peroxidase, phospholipid hydroperoxide glutathione peroxidase and glutathione S-transferase were significantly lower in Honghuadajinyuan than in Yunyan 87. Honghuadajinyuan and K326 had 119 differentially expressed proteins. 89 proteins of them covered metabolic pathways such as metabolism of xenobiotics by cytochrome P450. It was indicated that several proteins related to stress resistance such as proteinase inhibitor was much less in Yunyan 87 than in K326 and Honghuadajinyuan, and superoxide dismutase in Honghuadajinyuan was the lowest among the three varieties of tobacco. 【Conclusion】Results of the study revealed that the TMT coupled with 2D LC-MS/MS is a powerful method for isolating and identifying differentially expressed proteins in various tobacco varieties. Most of them are involved in photosynthesis, metabolism or stress resistance.

Key words: tobacco, differential proteomics, tandem mass tag, 2D LC-MS/MS

XU Ying, YAN Guo-quan, ZHANG Yang, YU Hong-xiu. Differential Proteomic Research of Three Varieties of Tobacco in China[J].Scientia Agricultura Sinica, 2016, 49(16): 3084-3097.

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[60] 杨建卿, 许大凤, 孔俊, 江彤. 不同烟草品种罹黑胫病后几种酶活性的变化. 合肥工业大学学报(自然科学版), 2005, 28(7): 816-819.

Yang J Q, Xu D F, Kong J, Jiang T. Change of activities of some enzymes in different tobacco varieties infected with Phytophthora nicotianae. Journal of Hefei University of Technology (Natural Sciences), 2005, 28(7): 816-819. (in Chinese)

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Differential Proteomic Research of Three Varieties of Tobacco in China

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