：Polyphenol oxidase (PPO) is a copper-containing metal enzyme, which plays an important role in tea processing. Five PPO genes were found in the tea genome database: CsPPO1, CsPPO2, CsPPO3, CsPPO4 (GenBank Access. No. GQ129142.1) and CsPPO5. The nucleic acid and amino acid sequences of five PPO genes were analyzed, and the physicochemical properties and structural functions of the PPO genes encoding protein were also predicted by bioinformatics method. The results showed that the full length of the five PPO genes coding sequences (CDS) was between 597βbp and 1β839βbp which could encode 198-612 amino acids. Phylogenetic tree analysis showed that CsPPO1, CsPPO3, CsPPO4 and CsPPO5 had close genetic relationships. The encoded proteins belonged to hydrophilic and unstable lipid binding proteins and had no signal peptides. Irregular curl was the main structural element of protein in which α-helix and β-strand were dispersed throughout the polypeptide chain & CsPPO1, CsPPO2, CsPPO3 might contain one or more transmembrane spirals. The five PPO proteins contained PPO1_DWL (pfam12142) conserved structural domains. It was predicted that the proteins encoded by CsPPO1, CsPPO3, CsPPO4 and CsPPO5 might contain tyrosinase functional domains. Real-time reverse transcription (qRT-PCR) analysis showed that the PPO genes showed different expression patterns among different tea cultivars.
[1] 伍梦瑶, 黄莹捷, 姚燕妮, 等. 勐库大叶种茶树多酚氧化酶粗酶的酶学性质[J]. 浙江大学学报(农业与生命科学版), 2017, 43(5): 579-588.
[2] 张跃进, 郝晓燕, 梁宗锁, 等. 莲藕多酚氧化酶基因(PPO)的克隆与表达分析[J]. 农业生物技术学报, 2011, 19(4): 634-641.
[3] 李桂琴, 李会宣, 许冬倩, 等. 鸭梨多酚氧化酶基因CDS区的克隆及表达[J]. 果树学报, 2008(4): 577-580.
[4] 时王珂, 孙奕玥, 舒志明, 等. 丹参多酚氧化酶基因的克隆及表达分析[J]. 西北植物学报, 2016, 36(9): 1735-1742.
[5] 王树军, 刘保华, 孙进华, 等. 荔枝多酚氧化酶基因启动子克隆与功能分析[J]. 果树学报, 2015, 32(3): 427-433, 524.
[6] Boss P K, Gardner R C, Janssen B J, et al.An apple polyphenol oxidase cDNA is up-regulated in wounded tissues[J]. Plant Molecular Biology, 1995, 27(2): 429-433.
[7] 王乃栋, 张丽霞, 向勤锃, 等. 茶树多酚氧化酶基因的生物信息学分析及原核表达[J]. 茶叶科学, 2011, 31(1): 33-39.
[8] 骆开明. 茶叶多酚氧化酶的基因克隆[J]. 山东农业科学, 2009(8): 9-12.
[9] 黄友谊. 茶树多酚氧化酶的基因克隆与原核表达[C]//中国茶叶学会. 2008茶学青年科学家论坛论文集. 2008: 9.
[10] 赵东, 刘祖生, 奚彪. 茶树多酚氧化酶基因的克隆及其序列比较[J]. 茶叶科学, 2001, 21(2): 94-98.
[11] Wu Y L, Pan L P, Yu S L, et al.Cloning, microbial expression and structure-activity relationship of polyphenol oxidases from Camellia sinensis[J]. Journal of Biotechnology, 2010, 145(1): 66-72.
[12] Liu J W, Huang Y Y, Ding J, et al.Prokaryotic expression and purification of Camellia sinensis polyphenol oxidase[J]. J Sci Food Agric, 2010, 90(14): 2490-2494.
[13] Wilkins M R, Gasteiger E, Bairoch A, et al.Protein identification and analysis tools in the ExPASy Server [M]//Walker J M. The Proteomics Protocols Handbook. New Jersey: Humana Press, 2005: 531.
[14] 孙奕玥. 丹参多酚氧化酶基因的克隆及其表达分析[D]. 杨凌: 西北农林科技大学, 2014.
[15] Cary J W, Lax A R, Flurkey W H.Cloning and characterization of cDNAs coding for Vicia faba polyphenol oxidase[J]. Plant Molecular Biology, 1992, 20(2): 245-253.
[16] Puranik S, Sahu P P, Mandal S N, et al. Comprehensive genome-wide survey, genomic constitution and expression profiling of the NAC transcription factor family in foxtail millet (Setaria italica L.) [J]. Plos One, 2013, 8(5): e64594. https://doi.org/10.1371/journal.pone.0064594
[17] 韦雪芳, 王冬梅, 刘思, 等. 信号肽及其在蛋白质表达中的应用[J]. 生物技术通报, 2006(6): 38-42.
[18] 罗勇, 欧淑琼, 陈丝, 等. 基于RNA-Seq的茶树CsMYB生物信息学分析[J]. 分子植物育种, 2017, 15(6): 2119-2125.
[19] 王乃栋. 茶多酚氧化酶基因的克隆及其工程菌的构建[D]. 泰安: 山东农业大学, 2012.
[20] Sommer A, Ne'Eman E, Steffens J C, et al. Import, targeting, and processing of a plant polyphenol oxidase[J]. Plant Physiology, 1994, 105(4): 1301-1311.
[21] 李吉. 膜蛋白跨膜螺旋预测的研究[D]. 上海: 上海交通大学, 2010.
[22] 李春美, 胡婉峰, 杨立, 等. 莲藕多酚氧化酶与底物或抑制剂相互作用的光谱学研究[J]. 分析试验室, 2010, 29(8): 13-17.
[23] 刘敬卫. 茶树多酚氧化酶的基因克隆与原核表达[D]. 武汉: 华中农业大学, 2009.
[24] Hunt M D, Eannetta N T, Yu H, et al.cDNA cloning and expression of potato polyphenol oxidase[J]. Plant Molecular Biology, 1993, 21(1): 59-68.
[25] Shahar T, Hennig N, Gutfinger T, et al.The tomato 66.3-kD polyphenoloxidase gene: molecular identification and developmental expression[J]. Plant Cell, 1992, 4(2): 135-147.
[26] 刘仲华, 黄建安. 二十个茶树品种(品系)红碎茶适制性的研究[J]. 中国茶叶加工, 1998(2): 29-32.
[27] 闫洪波, 李桂琴, 宋亚伟, 等. 5种蔷薇科树种多酚氧化酶比较基因组学分析[J]. 热带作物学报, 2017, 38(2): 320-327.
[28] Marusek C M, Trobaugh N M, Flurkey W H, et al.Comparative analysis of polyphenol oxidase from plant and fungal species[J]. J Inorg Biochem, 2006, 100(1): 108-123.
[29] Flurkey W H, Inlow J K.Proteolytic processing of polyphenol oxidase from plants and fungi[J]. J Inorg Biochem, 2008, 102(12): 2160-2170.
[30] 潘宇, 王坤波, 徐仲溪, 等. 茶叶多酚氧化酶的序列分析与结构预测[J]. 茶叶科学, 2008, 28(3): 157-165.
[31] 孙晓玲, 蔡晓明, 马春雷, 等. 茉莉酸甲酯和机械损伤对茶树叶片多酚氧化酶时序表达的影响[J]. 西北植物学报, 2011, 31(9): 1805-1810.
[32] 母洪娜, 王良桂, 孙陶泽. 桂花多酚氧化酶(PPO)基因的克隆及表达分析[J]. 分子植物育种, 2017, 15(2): 441-446.
[33] Stewart R J, Sawyer B J B, Bucheli C S, et al. Polyphenol oxidase is induced by chilling and wounding in pineapple[J]. Australian Journal of Plant Physiology, 2001, 28(3): 181-191.
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