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標題: 利用SWATH質譜定量技術於水稻葉色突變株之蛋白質體研究
SWATH-based comparative proteomics analysis of leaf color mutant in Oryza sativa
作者: 蔡鴻樹
Hung-Shu Tsai
關鍵字: 水稻;葉色突變株;蛋白質體;oryza sativa;proteomics;SWATH;PAL
引用: 1. Evans, J. R. Improving Photosynthesis. Plant physiology 162 (2013) 1780-1793. 2. McCabe, M. S., Garratt, L. C., Schepers, F., Jordi, W. J.; Stoopen, G. M., Davelaar, E., van Rhijn, J. H., Power, J. B., Davey, M. R. Effects of PSAG12-IPT Gene Expression on Development and Senescence in Transgenic Lettuce Plant physiology. 127 (2001) 505-516. 3. Juan, A.P. , Elizabete, C. , Carmen, H. , Jaume, F. and Jeroni, G. Acclimation of Biochemical and Diffusive Components of Photosynthesis in Rice, Wheat, and Maize to Heat and Water Deficit: Implications for Modeling Photosynthesis. Plant Science 7 (2016) 1719. 4. Gammulla, C. G., Pascovici, D., Atwell, B. J., Haynes, P. A. Differential metabolic response of cultured rice (Oryza sativa) cells exposed to high-and low-temperature stress. Proteomics. 10 (2010) 3001-3019. 5. Lanlan, W. , Jun, W. , Yanxia, L. , Xiaodi, G. , Jianlong, X. , Dongzhi, L. , Yanjun, D. The Rice Pentatricopeptide Repeat Gene TCD10 is Needed for Chloroplast Development under Cold Stress. Rice 9 (2016) 67. 6. Wang, S., Uddin, M. I., Tanaka, K., Yin, L., Shi, Z., Qi, Y., Mano, J., Matsui, K., Shimomura, N., Sakaki, T., Deng, X., Zhang, S. Maintenance of chloroplast structure and function by overexpression of the rice MONOGALACTOSYLDIACYLGLYCEROL SYNTHASE gene leads to enhanced salt tolerance in tobacco. Plant physiology. 165 (2014) 1144-1155. 7. Zhang, H., Han, B., Wang, T., Chen, S. X., Li, H. Y., Zhang, Y. H., Dai, S. Mechanisms of plant salt response insights from proteomics. J. Proteome Res. 11 (2012) 49-67. 8. Siena, I. 2D Electrophoresis: From Protein Maps to Genomes. Proceedings of the International Meeting. Electrophoresis. 16 (1995) 1077-1322. 9. Megger, D. A., Bracht, T., Meyer, H. E., Sitek, B. Label-free quantification inclinical proteomics. Biochim. Biophys. Acta. 1384 (2013) 1581-1590. 10. Lin, D. G., Chou, S. Y., Wang, A. Z., Wang, Y. W., Kuo, S. M., Lai, C. C., Chen, L. J., Wang, C. S. A proteomic study of rice cultivar TNG67 and its high aroma mutant SA0420. Plant Science. 214 (2014) 20-28. 11. Chang, T. S., Liu, C. W., Lin, Y. L., Li, C. Y., Wang, A. Z., Chein, M. W., Wang, C. S., Lai, C. C. Mapping and comparative proteomic analysis of the starch biosynthetic pathway in rice by 2D PAGE/MS. Plant Mol. Biol. 95 (2017) 333-343. 12. Liu, C. W., Hsu, Y. K., Cheng, Y. H., Yen, H.C., Wu, Y. P., Wang, C. S., Lai, C. C. Proteomic analysis of salt-responsive ubiquitin-related proteins in rice roots. Rapid Commun. Mass Spectrom. 26 (2012) 1649-1660. 13. Liu, C. W., Chang, T. S., Hsu, Y. K., Wang, A. Z., Yen, H. C., Wu, Y. P., Wang, C. S., Lai, C. C. Comparative proteomic analysis of early salt stress-responsive proteins in roots and leaves of rice. Proteomics. 15 (2014) 1759-1775. 14. Zhao, Y., Chen, P., Lin, L., Harnly, J., Yu, L.L., Li, Z. Tentative identification, quantitation, and principal component analysis of green pu-erh, green, and white teas using UPLC/DAD/MS. Food Chem. 126 (2011) 1269-1277. 15. Bondarenko, P. V., Chelius, D., Shaler, T. A. Identification and Relative Quantitation of Protein Mixtures by Enzymatic Digestion Followed by Capillary Reversed-Phase Liquid Chromatography-Tandem Mass Spectrometry. Anal Chem. 74 (2002) 4741-4749. 16. Liu, H., Sadygov, R. G., Yates, J. R., 3rd. A model for random sampleing and estimation of relative protein abundance in shotgun proteomics. Anal Chem. 76 (2004) 4193-4201. 17. Haverland, N. A., Fox, H. S., Ciborowski, P. J. Quantitative Proteomics by SWATH-MS Reveals Altered Expression of Nucleic Acid Binding and Regulatory Proteins in HIV-1-Infected Macrophages. Proteome Res. 13 (2014) 2109-2119. 18. Kanchan,V., Keith, A., Sarah, R., Carlos, S., Suchismita, S., Jose, A. A., Alejandra, P., Sebastian, E.I., David, K., Murray, D.M., Gregory, E.R. Applying SWATH Mass Spectrometry to Investigate Human Cervicovaginal Fluid During the Menstrual Cycle. biology of reproduction 39 (2015) 1–10. 19. Zhang, H., He, D., Yu, J., Li, M., Damaris, R.N., Gupta, R., Kim, S.T., Yang, P. Analysis of dynamic protein carbonylation in rice embryo during germination through AP-SWATH. Proteomics. 16 (2016) 989–1000. 20. Suwannatrai, K., Suwannatrai, A., Tabsripair,P., Welbat, J.U., Tangkawattana, S., Cantacessi, C., Mulvenna, J., Tesana, S., Loukas, A., Sotillo, J. Differential Protein Expression in the Hemolymph of Bithynia siamensis goniomphalos Infected with Opisthorchis viverrini. PLOS Neglected Tropical Diseases 10 (2016) 1-20. 21. Fabre, B., Korona, D., Nightingale, D.J., Russell, S., Lilley, K.S. SWATH-MS data of Drosophila melanogaster proteome dynamics during embryogenesis. Data In Brief 9 (2016) 991-995. 22. Fabre, B., Korona, D., Nightingale, D.J., Russell, S., Lilley, K.S. SWATH-MS data of Drosophila melanogaster proteome dynamics during embryogenesis. Data In Brief 9 (2016) 771-775. 23. Losensky, G., Jung, K., Urlaub, H., Pfeifer, F., Fröls, S., Lenz, C. Shedding light on biofilm formation of Halobacterium sanlinarum R1 by SWATH-LC/MS/MS analysis of planktonic and sessile cells. Proteomics 17 (2017) 1-13. 24. Stauch, K.L., Villeneuve, L.M., Purnell, P.R., Pandey, S., Guda, C., Fox, H.S. SWATH-MS proteome profiling data comparison of DJ-1, Parkin, and PINK1 knockout rat striatal mitochondria. Data In Brief 9 (2016) 589-593. 25. Sanda, M., Zhang, L., Edwards, N.J., Goldman, R. Site-specific analysis of changes in the glycosylation of proteins in liver cirrhosis using data-independent workflow with soft fragmentation. Analytical and Bioanalytical Chemistry. 2 (2017) 619-627. 26. Zhang, X., Walsh, T., Atherton, J. J., Kostner, K., Schulz, B., Punyadeera, C. Identification and Validation of a Salivary Protein Panel to Detect Heact Failure Early. Theranostics. 18 (2017) 4350-4358. 27. Liu, Y., Borel, C., Li, L., Müller, T., Williams, E. G., Germain, P. L., Buljan, M., Sajic, T., Boersema, P. J., Shao, W., Faini, M., Testa, G., Beyer, A., Antonarakis, S. E., Aebersold, R. Systematic proteome and proteostasis profiling in human Trisomy 21 fibroblast cells. Nat Commun. 1 (2017) 1212. 28. Dalla, P. E., Manfredi, M., Brandi, J., Buzzi, A., Conte, E., Pacchiana, R., Cecconi, D., Marengo, E., Donadelli, M. J. Trichostatin A alters cytoskeleton and energy metabolism of pancreatic adenocarcinoma cells. Cell Biochem. 1 (2017) 1-12. 29. Chaudhari, P. R., Charles, S. E., D'Souza, Z. C., Vaidya, M. M. Hemidesmosomal linker proteins regulate cell motility, invasion and tumorigenicity in oral squamous cell carcinoma derived cells. Cell Res. 2 (2017) 125-137. 30. Dhakarey, R., Raorane, M. L., Treumann, A., Peethambaran, P. K., Schendel, R. R., Sahi, V. P., Hause, B., Bunzel, M., Henry, A., Kohli, A., Reimann, M. Physiological and Proteomic Analysis of the Rice Mutant cmp2 Suggests a Negative Regulatory Role Jasmonic Acid in Drought Tolerance. Front Plant Sci. 8 (2017) 1903. 31. Yang, A., Yu, L., Chen, Z., Zhang, S. Shi, J., Zhao, X., Yang, Y., Hu, D., Song, B. Label-Free Quantitative Proteomic Analysis of Chitosan Oligosaccharide-Treated Rice Infected with Southern Rice Black-Streaked Dwarf Virus. Viruses. 9 (2017) 5. 32. Yunqi, W., Mehdi, M., Dand, P., Joel, M. C., Brain, J. A., Paul, A. H. Quantitative proteomic analysis of two different rice varieties reveals that drought tolerance is correlated with reduced abundance of photosynthetic machinery and increased abundance of ClpD1 protease. J. Proteomics. 143 (2016) 73-82. 33. Zhang, H., Dongli, H., Jianlan, Y., Ming, L., Rebecca, N. D., Ravi, G., Sun, T. K., Pingfang, Y. Analysis of dynamic protein carbonylation in rice embryo during germination through AP-SWATH. Proteomics 16 (2016) 989-1000. 34. Chen, L., Huang, Y., Xu, M., Cheng, Z., Zheng, J. Proteomic Analysis Reveals Coordinated Regulation of Anthocyanin Biosynthesis through Signal Transduction and Sugar Metabolism in Black Rice Leaf. Int. J. Mol. Sci. 18 (2017) 12. 35. Galland, M., He, D., Lounifi, I., Arc, E., Clement, G., Balzergue, S., Huguet, S., Cueff, G., Godin, B., Collet, B., Granier, F., Morin, H., Tran, J., Valot, B., Rajjou, L. An Intergrated 'Multi-Omics' Comparison of Embyro and Endosperm Tissue-Specific Features and Their Impact on Rice Seed Quality. Front Plant Sci. 8 (2017) 1984. 36. Xue, C., Liu, S., Chen, C., Zhu, J., Yang, X., Zhou, Y., Guo, R., Liu, X., Gong, Z. Global Proteome Analysis Links Lysine Acetylation to Diverse Functions in Oryza Sativa. Proteomics 18 (2018) 1. 37. Mu, Q., Zhang, W., Zhang, Y., Yan, H., Matsui, T., Tian, X., Yang, P. iTRAQ-Based Quantitative Proteomics Analysis on Rice Anther Responding to High Temperature. Int. J. Mol. Sci. 18 (2017) 9. 38. Zhu, F. Y., Chen, M. X., Su, Y. W., Xu, X., Ye, N. H., Cao, Y. Y., Sheng, L., Liu, T. Y., Li, H. X., Wang, G. Q., Jin, Y., Gu, Y. H., Chan, W. L., Lo, C., Peng, X., Zhu, G., Zhang, J. SWATH-MS Quantitative Analysis of Proteins in the Rice Inferior and Superior Spikelets during Grain Filling. Front Plant Sci. 7 (2016) 1926. 39. Collins, B.C., Hunter, C.L., Liu, Y., Schilling, B., Rosenberger, G., Bader, S.L., Chan, D.W., Gibson, B.W., Gingras, A.C., Held, J.M., Hirayama-Kurogi, M., Hou, G., Krisp, C., Larsen, B., Lin, L., Liu, S., Molloy, M.P., Moritz, R.L., Ohtsuk S., Schlapbach, R., Selevsek, N., Thomas, S.N., Tzeng, S.C., Zhang, H., Aebersold, R. Multi-laboratory assessment of reproducibility, qualitative performance of SWATH-mass spectrometry. Nat. Commun. 1 (2017) 291. 40. Boyer, P.D. The binding change mechanism for ATP synthase - some probabilities and possibilities. Biochim. Biophys. Acta 1140 (1993) 215–250. 41. Abrahams, J.P., Leslie, A.G.W., Lutter, R. and Walker, J.F. Structure at 2.8 Å resolution of F1-ATPase from bovine heart mitochondria. Nature 375 (1994) 621–628. 42. Blair, A., Ngo, L., Park, J., Paulsen, I.T. and Saier, M.H., Jr. Phylogenetic analyses of the homologous transmembrane channel-forming proteins of the FoF1-ATPases of bacteria, chloroplasts and mitochondria. Microbiology 142 (1996) 17–32. 43. Noji, H., Yasuda, R., Yoshida, M. and Kinosita, K., Jr. Direct observation of the rotation of F1-ATPase. Nature 386 (1997) 299–302. 44. Knaff D, B., Malkin, R., Myron, J.C., Stoller, M. The role of plastoquinone and beta-carotene in the primary reaction of plant photosystem II. Biochim. Biophys. Acta 459 (1977) 402-411. 45. Guskov, A., Kern, J., Gabdulkhakov, A., Broser, M., Zouni, A., Saenger, W. Cyanobacterial photosystem II at 2.9-A resolution and the role of quinones, lipids, channels and chloride. Nat. Struct. Mol. Biol. 16 (2009) 334-342. 46. Karplus, P.A., Daniels, M.J., Herriott, J.R. Atomic structure of ferredoxin-NADP+ reductase: prototype for a structurally novel flavoenzyme family. Science 251 (1991) 60-66. 47. Johansson, P., Hederstedt, L. Organization of genes for tetrapyrrole biosynthesis in gram--positive bacteria. Mircobiology 145 (1999) 529-538. 48. Kohno, H., Furukawa, T., Yoshinaga, T., Tokunaga, R., Taketani, S. Coproporphyrinogen oxidase. Purification, molecular cloning, and induction of mRNA during erythroid differentiation. J Biol Chem 268 (1993) 21359-21363. 49. Tanaka, R., Oster, U., Kruse, E., Rudiger, W., Grimm, B. Reduced activity of geranylgeranyl reductase leads to loss of chlorophyll and tocopherol and to partially geranylgeranylated chlorophyll in transgenic tobacco plants expressing antisense RNA for geranylgeranyl reductase. Plant Physiol 120 (1999) 695-704. 50. Keller, Y., Bouvier, F., d'Harlingue, A., Camara, B. Metabolic compartmentation of plastid prenyllipid biosynthesis--evidence for the involvement of a multifunctional geranylgeranyl reductase. Eur J Biochem 251 (1998) 413-417. 51. Addlesee, H.A., Hunter, C.N. Rhodospirillum rubrum possesses a variant of the bchP gene, encoding geranylgeranyl-bacteriopheophytin reductase. J Bacteriol 184 (2002) 1578-1586. 52. Ohta, S., Kagawa, Y. Human F1-ATPase: molecular cloning of cDNA for the beta subunit. J. Biochem. 99 (1986) 135-141. 53. Takase, K., Kakinuma, S., Yamato, I., Konishi, K., Igarashi, K., Kakinuma, Y. Sequencing and characterization of the ntp gene cluster for vacuolar-type Na(+)-translocating ATPase of Enterococcus hirae. J. Biol. Chem. 269 (1994) 11037-11044. 54. Cipriano, D.J., Wang, Y., Bond, S., Hinton, A., Jefferies, K.C., Qi, J., Forgac, M. Structure and regulation of the vacuolar ATPases. Biochim Biophys Acta 1777 (2008) 599-604. 55. Jefferies, K.C., Cipriano, D.J., Forgac, M. Function, structure and regulation of the vacuolar (H+)-ATPases. Arch Biochem Biophys 476 (2008) 33-42. 56. Nishikimi, M., Ohta, S., Suzuki, H., Tanaka, T., Kikkawa, F., Tanaka, M., Kagawa, Y., Ozawa, T. Nucleotide sequence of a cDNA encoding the precursor to human cytochrome c1. Nucleic Acids Res. 16 (1988) 3577. 57. Galeotti, F., Barile, E., Curir, P., Dolci, M., Lanzotti, V. Flavonoids from carnation (Dianthus caryophyllus) and their antifungal activity. Phytochemistry Letters 1 (2008) 44-48. 58. Davey, M.P., Bryant, D.N., Cummins, I., Gates, P., Ashenden, T.W., Baxter, R., Edwards, R. Effects of elevated CO2 on the vasculature and phenolic secondary metabolism of Plantago maritima. Phytochemistry 65 (2004) 2197-2204. 59. Lebo, S.E.Jr., Gargulak, J.D., McNally, T.J. Lignin Kirk Othmer Encyclopedia of Chemical Technology (2001) 60. Boerjan, W., Ralph, J., Baucher, M., Lignin biosynthesis. Annu. Rev. Plant Biol. 54 (2003) 519-549. 61. Camm, E.L., Towers, G.H.N. Phenylalanine ammonia lyase Phytochemistry 12 (1973) 961-973. 62. Hahlbrock, K., Grisebach, H. Enzymic Controls in the Biosynthesis of Lignin and Flavonoids Annual Review of Plant Physiology 30 (1979) 105-130. 63. Dixon, R.A., and Paiva, N.L. Stress-induced phenylpropanoid metabolism. Plant Cell 7 (1995) 1085-1097. 64. Mauch-Mani, B., and Slusarenko, A.J. Production of salicylic acid precursors is a major function of phenylalanine ammonia-lyase in the resistance of arabidopsis to Peronospora parasitica. Plant Cell 8 (1996) 203-212. 65. Ehness, R., Ecker, M., Godt, D.E., and Roitsch, T. Glucose and stress independently regulate source and sink metabolism and defense mechanisms via signal transduction pathways involving protein phosphorylation. Plant Cell 9 (1997) 1825-1841. 66. Sarkissian, C.N.; Gámez, A. Phenylalanine ammonia lyase, enzyme substitution therapy for phenylketonuria, where are we now? Molecular Genetics and Metabolism 86 (2005) 22-26. 67. Gámez, A., Wang, L., Straub, M., Patch, M.G., Stevens, R.C. Toward PKU enzyme replacement therapy: PEGylation with activity retention for three forms of recombinant phenylalanine hydroxylase. Molecular Therapy 9 (2004) 124-129. 68. Lenke, R.R., Levy, H.L. Maternal phenylketonuria--results of dietary therapy American Journal of Obstetrics and Gynecology 142 (1982) 548-553. 69. Fritz, R.R., Hodgins, D.S., Abell, C.W. Phenylalanine ammonia-lyase. Induction and purification from yeast and clearance in mammals. The Journal of Biological Chemistry 251 (1976) 4646-4650. 70. Tiller, N., Bock, R. The translational apparatus of plastids and its role in plant development. Molecular Plant 7 (2014) 1105-1120. 71. Tuanzhang, Y., Gang, P., Han, L., Jian, W., Yongpeng, L., Zhenxing, Z., Tingdong, F., Yongming, Z. The chloroplast ribosomal protein L21 gene is essential for plastid development amd embryogenesis in Arabidopsis. Planta 235 (2012) 901-921. 72. Isidora, R., Luca, T., Fabio, R., Simona, M., Mathias, P., Peter, J., Martin, K., Dario, L., Paolo, P. Versatile roles of Arabidopsis plastid ribosomal proteins in plant growth and development. The Plant Journal 72 (2012) 922-934. 73. Lin, D., Jiang, Q., Zheng, K., Chen, S., Zhou, H., Gong, X., Xu, J., Teng, S., Dong, Y. Mutation of rice ASL2 gene encoding plastid ribosomal protein L21 causes chloroplast developmental defects and seddling death. Plant Biology 17 (2015) 599-607. 74. Anne-Sophie, P., Tina, K., Rana, H., Alexis, D., Myriam, C., Janice, A.E., Godelieve, G., Diana, F. Transcriptomic and histological responses of African rice (Oryza glaberrima) to Meloidogyne graminicola provide new insights into root-knot nematode resistance in monocots. Annals of Botany 118 (2017) 885-899. 75. Xiang, L., Yongmei, H., Chunmei, X., Yanqun, Z., Fangdong, Z., Xinyue, M., Yang, X., Yuan, L. Effects of UV-B radiation on the infectivity of Magnaporthe oryzae and rice disease-resistant physiology in Yuanyang terraces. Photochem. Photobiol. Sci. 17 (2018) 8-17. 76. Afroz, R., Muhammad, S., Kamran, M., Fauzia, Y.H., Humaira, Y., Saqib, M., Muhammad, N.H. Antagonistic Bacillus spp. reduce blast incidence on rice and increase grain yield under field conditions. Microbiological Research 208 (2018) 54-62. 77. Afroz, R., Zahra, J., Faluk, S., Fauzia, Y.H., Muhammad, N.H. Bacillus spp., a bio-control agent enhances the activity of antioxidant defense enzymes in rice against Pyricularia oryzae. PLoS ONE 12 (2017) 11. 78. Yongqiang, H., Pei, L., Shaolong, G., Lang, Y., Lizhang, W., Maolin, H. Defense Responses in Rice Induced by Silicon Amendment against Infestation by the Leaf Folder Cnaphalocrocis medinalis. PLoS ONE 11 (2016) 4. 79. Kanehisa, Furumichi, M., Tanabe, M., Sato, Y., Morishima, K. KEGG: new perspectives on genomes, pathways, diseases and drugs. Nucleic Acids Res. 45 (2017) 353-361. 80. Kanehisa, M., Sato, Y., Kawashima, M., Furumichi, M., Tanabe, M. KEGG as a reference resource for gene and protein annotation. Nucleic Acids Res. 44 (2016) 457-462. 81. Kanehisa, M., Goto, S. KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res. 28 (2000) 27-30.
光合作用為整個植物的主要能量來源,會影響植物是否可以存活下去;此作用與葉片的顏色息息相關,當植物葉片轉黃時,會因為無法形光合作用,導致死亡。而水稻葉色突變株 SA0405、SA0407 和 SA0408為台農67號 (TNG67) 經疊氮化鈉誘變之水稻葉色突變株,其外表性狀及生理狀況有明顯不同,株高較矮小且葉片顏色偏黃偏淡,卻能完整生長、抽穗及成熟,但其產生原因尚未明瞭。近年來,質譜發展迅速,其中蛋白質體學技術已應用於許多領域,包含植物性狀研究。2012 年Aebersold 等人以開發出非標定定量技術 Sequential window acquisition of all theoretical fragment ion spectra (SWATH) 之質譜掃描模式,其能偵測特定視窗內的所有母離子,加以撞碎後,收集其所有碎片離子之訊號,對於蛋白質之定量分析是一大幫助。因此,本實驗以非標定定量技術 SWATH 搭配超高效能液相層析儀 (ultra high performance liquid chromatography, UHPLC) 串聯四極棒飛行時間式質譜儀 (quadrupole/time-of-flight mass spectrometry, QTOF MS),探討葉色突變株 SA0405、SA0407 和 SA0408 與 TNG67 葉片之插秧後第 45 天、最高分蘗期、抽穗期與成熟期之蛋白質體表現差異,期望能了解此外表性狀、生理狀況之原因。在 16 個樣品中,總共鑑定到 2408 個蛋白。在這些蛋白中,則以表現量比值 > 1.5或 < 0.67 作為顯著差異標準 (突變株/TNG67)。而藉由 KEGG pathway 處理後,發現突變株中 Phenylalanine ammonia-lyase (PAL) 有大量表現,而 PAL 則會形成類黃酮、苯丙素及木質素等激素的前驅物,因此懷疑是此蛋白之變化導致突變株獨特的性狀。而 PAL 為抗逆境蛋白,在本實驗的結果中發現突變株葉綠體 ribosome 中的 L21 (G23) 及 L27 (G36) 蛋白大量下降。而這兩個蛋白若大量下降時,會影響葉綠體內重要蛋白之合成。因此就此產生逆境。從此結果得知,本實驗成功藉由 SWATH 質譜定量技術,找出葉色突變株與 TNG67 在外表性狀及生理狀況為何有如此差異的原因。

Photosynthesis provided the main energy in plants. Plants could not survive without photosynthesis. Photosynthesis was highly related to the color of leaves. The leaf color mutants SA0405, SA0407 and SA0408 used in this study were induced from a wild-type rice, TNG67, by sodium azide mutagenesis; therefore, TNG 67 and color mutants share the same genetic background. The phenotypes and physiology of these color mutants were obviously different from TNG67. Surprisingly, they have similar photosynthesis efficiency and life cycle. But the molecular mechanisms involved in these phenomenon is not clear. In recent years, mass spectrometry develop rapidly. Proteomics have been applied in many researchs, include the phenotypes and physiology of plant. In 2012, Aebersold etc. have developed a new label-free quantitative proteomics method, Sequential window acquisition of all theoretical fragment ion spectra (SWATH). SWATH data was acquired by repeatedly cycling through sequential mass windows over the whole chromatographic elution range generating a complete recording of all analytes in the sample. Thus, this paper will use SWATH method with ultra high performance liquid chromatography (UHPLC) and quadrupole/time-of-flight mass spectrometry (QTOF MS) to research the proteomics between leaf color mutants and wild-type rice in 45 days after transplant, tillering, spiking, mature period. Expect the results may be helpful in further elucidating the molecular mechanisms between leaf color mutants and wild-type rice. In the 16 samples, 2408 proteins were generated. Proteins with a fold change of > 1.5 or < 0.67 (mutants/TNG67) considered as differentially expressed proteins in this research. After analysis by KEGG pathway, find out the quantity of Phenylalanine ammonia-lyase (PAL) in leaf color mutants bigger than the quantity in wild-type rice. PAL will produce flavonoids, phenylpropanoids, lignins and other enzymes. Thus, we doubt this protein highly related to the phenotypes and physiology of leaf color mutants. PAL is a protein for confrontation in adversity. In our results, we also find L21 and L27 down expression in leaf color mutants, which are the proteins in chloroplast ribosome. If those two proteins have down-expression, the proteins in chloroplast will not be synthesis. This is the reason for adversity. Finally, we use SWATH to find out the relationship between leaf color mutants and wild types rice successfully.
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