茶树儿茶素合成相关MYB转录因子生物信息学分析

doi:10.13305/j.cnki.jts.2018.02.007

摘要/Abstract

摘要： 从茶树（Camellia sinensis）转录组数据库中获得6个与儿茶素合成相关的MYB转录因子,对其进行蛋白理化性质与结构功能预测、核定位信号和蛋白保守结构域分析,通过同源分析构建系统发育树。结果表明,与儿茶素合成相关的6个MYB蛋白都属于亲水性的非分泌蛋白,定位在叶绿体、线粒体和细胞核上,其空间结构主要是α-螺旋和β-转角;保守结构域的分析发现除了comp159173_c0基因,其余5个都属于SANT超家族基因成员。系统进化树显示6个MYB蛋白形成4个分支,显示出各自间较远的亲缘关系。差异表达分析的结果进一步证实了6个MYB转录因子与儿茶素合成代谢的相关性。

关键词: 儿茶素, MYB转录因子, 生物信息学

Abstract: Six MYB transcription factors involved in the biosynthesis of catechins were obtained from the tea plant (Camellia sinensis) transcriptome database. Bioinformatic analysis including the physicochemical properties and structural functions of proteins, nuclear localization signal (NLS), protein conserved domain (PCD) and phylogenetic tree analysis were performed. The results demonstrated that the 6 MYB proteins involved in the biosynthesis of catechins belong to hydrophilic non-secretory proteins and are predicted to be located in the chloroplasts, mitochondria and nucleus. The main spatial structures are α-helixes and β-turns. Except the comp159173_c0, the other 5 genes belong to SANT super-family group based on the PCD analysis. The phylogenetic tree analysis shows that 6 MYB proteins are classified into 4 groups, indicating a distant relationship between them. Differential gene expression analysis further confirmed the close correlation between the 6 transcription factors and the catechin biosynthesis.

Key words: catechins, MYB transcription factors, bioinformatics

中图分类号:

张玥, 胡雲飞, 王树茂, 柯子星, 林金科. 茶树儿茶素合成相关MYB转录因子生物信息学分析[J]. 茶叶科学, 2018, 38(2): 162-173. doi: 10.13305/j.cnki.jts.2018.02.007.

ZHANG Yue, HU Yunfei, WANG Shumao, KE Zixing, LIN Jinke. Bioinformatic Analysis of MYB Transcription Factors Involved in Catechins Biosynthesis in Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2018, 38(2): 162-173. doi: 10.13305/j.cnki.jts.2018.02.007.

参考文献 41

[1]	宛晓春. 茶叶生物化学[M]. 3 版. 北京: 中国农业出版社, 2003: 9-18.
[2]	Cabrera C, Aatacho R,Gimenez R.Beneficial effects of green tea-a review[J]. J Am Coll Nutr, 2006, 25(2): 79-99.
[3]	Lee M M, Schiefelbein J.Cell pattern in the Arabidopsis root epidermis determined by lateral inhibition with feedback[J]. Plant Cell, 2002, 14(3): 611-618.
[4]	Zhang Y, Cao G, Qu LJ, et al.Characterization of Arabidopsis MYB transcription factor gene AtMYB17 and its possible regulation by LEAFY and AGL15[J]. J Genet. Genomics, 2009, 36(2): 99-107.
[5]	Fornalé S, Lopez E, Salazar-Henao J E, et al. AtMYB7: a new player in the regulation of UV-sunscreens in Arabidopsis thaliana[J]. Plant Cell Physiol, 2014, 55(3): 507-516.
[6]	Raffaele S, Rivas R, Roby D.An essential role for salicylic acid in AtMYB30-mediated control of the hypersensitive cell death program in Arabidopsis[J]. FEBS Lett, 2006, 580(14): 3498-3504.
[7]	Li M, Dong XJ, Peng JQ, et al.De novo transcriptome sequencing and gene expression analysis reveal potential mechanisms of seed abortion in dove tree (Davidia involucrata Baill.)[J]. BMC Plant Biol, 2016, 16(82): 1-21.
[8]	Park MY, Kang JY, Kim SY.Overexpression of AtMYB52 confers ABA hypersensitivity and drought tolerance[J]. Mol Cells, 2011, 31(5): 447-454.
[9]	Vom Endt D, Kijne JW, Memelink J.Transcription factors controlling plant secondary metabolism: what regulates the regulators[J]. Phytochemistry, 2002, 61(2): 107-114.
[10]	Song SS, Qi TC, Huang H, et al.The Jasmonate-ZIM domain proteins interact with the R2R3-MYB transcription factors MYB21 and MYB24 to affect Jasmonate-regulated stamen development in Arabidopsis[J]. Plant Cell, 2011, 23(3): 1000-1013.
[11]	Schwinn KE, Murray MR, Bradley JM, et al.MYB and bHLH transcription factor transgenes increase anthocyanin pigmentation in petunia and lisianthus plants, and the petunia phenotypes are strongly enhanced under field conditions[J]. Front Plant Sci, 2014, 5: 603.
[12]	牛义岭, 姜秀明, 许向阳. 植物转录因子MYB基因家族的研究进展[J]. 分子植物育种, 2016, 14(8): 2050-2059.
[13]	成浩, 李素芳. 茶树中的类黄酮物质及其生物合成途径[J]. 中国茶叶, 1999, 6(1): 6-8.
[14]	马春雷, 乔小燕, 陈亮. 茶树无色花色素还原酶基因克隆及表达分析[J]. 茶叶科学, 2010, 30(1): 27-36.
[15]	马春雷, 陈亮. 茶树黄酮醇合成酶基因的克隆与原核表达[J]. 基因组学与应用生物学, 2009, 28(3): 433-438.
[16]	Mamati G E, Liang Y,Lu J.Expression of basic genes involved in tea polyphenol synthesis in relation to accumulation of catechins and total tea polyphenols[J]. Journal of the Science of Food and Agriculture, 2006, 86: 459-464.
[17]	马春雷. 茶树查尔酮异构酶、黄酮醇合成酶和无色花色素还原酶等基因的克隆与表达分析[D]. 北京: 中国农业科学院, 2007.
[18]	邢文, 金晓玲. 调控植物类黄酮合成的MYB转录因子研究进展[J]. 分子植物育种, 2015, 13(3): 689-696.
[19]	Aharoni A, De Vos CH, Wein M, et al.The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco[J]. Plant J, 2001, 28(3): 319-332.
[20]	Ballester AR, Molthoff J, de Vos R, et al. Biochemical and molecular analysis of pink tomatoes: deregulated expression of the gene encoding transcription factor SlMYB12 leads to pink tomato fruit color[J]. Plant Physiol, 2010, 152(1): 71-84.
[21]	Espley RV, Hellens RP, Putterill J, et al.Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10[J]. Plant J, 2007, 49(3): 414-427.
[22]	Jung CS, Grifliths HM, De Jong DM, et al.The potato developer(D) locus encodes an R2R3MYB transcription factor that regulates expression of multiple anthocyanin structural genes in tuber skin[J]. Theor Appl Genet, 2009, 120(1): 45-57.
[23]	Yoshida K, Iwasaka R, Kaneko T, et al.Functional differentiation of lotus japonicus TT2s, R2R3-MYB transcription factors comprising a multigene family[J]. Plant Cell Physiol, 2008, 49(2): 157-169.
[24]	Akagi T, Ikegami A, Tsujimoto T, et al.DkMyb4 is a Myb transcription factor involved in proanthocyanidin biosynthesis in persimmon fruit[J]. Plant Physiol, 2009, 151(4): 2028-2045.
[25]	Mellway R D, Tran L T, Prouse M B, et al.The wound-, pathogen-, and ultraviolet B-responsive MYB134 gene encodes an R2R3 MYB transcription factor that regulates proanthocyanidin synthesis in poplar[J]. Plant Physiology, 2009, 150(2): 924-941.
[26]	Mehrtens F, Kranz H, Bednarek P, et al.The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis[J]. Plant Physiol, 2005, 138(2): 1083-1096.
[27]	Stracke R, Ishihara H, Huep G, et al.Differential regulation of closely related R2R3-MYB transcription factors controls fiavonol accumulation in different parts of the Arabidopsis thaliana seedling[J]. Plant J, 2007, 50(4): 660-677.
[28]	Czemmel S, Stracke R, Weisshaar B, et al.The grapevine R2R3-MYB transcription factor VvMYBF1 regulates fiavonol synthesis in developing grape berrie[J]. Plant Physiol, 2009, 151(3): 1513-1530.
[29]	Bovy A, de Vos R, Kemper M, et al. High-flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1[J]. Plant Cell, 2002, 14(10): 2509-2526.
[30]	陈玮, 赵贤倩, 吴亚会, 等. 茶树MYB7转录因子功能的研究[J]. 安徽农业大学学报, 2015, 42(6): 841-848.
[31]	孙彬妹. 茶树MYB转录因子CsAN1调控花青素的作用机制研究[D]. 广州: 华南农业大学, 2016.
[32]	葛国平, 吴亮宇, 张少雄, 等. 茶树芽叶儿茶素含量与转录因子MYB、bHLH关联分析[J]. 广东农业科学, 2014, 9: 28-32.
[33]	薛庆中. DNA和蛋白质序列数据分析工具[M]. 3版. 北京: 科学出版社, 2010: 75-106.
[34]	Mortazavi A, Williams B, Mccue K, et al.Apping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nature methods, 2008, 5(7): 621-628.
[35]	罗勇, 欧淑琼, 陈丝, 等. 基于RNA-Seq的茶树Cs-MYB生物信息学分析[J]. 分子植物育种, 2017, 15(6): 2119-2125.
[36]	谌章舟. CoREST SANT结构域蛋白与非编码小dsRNA相互作用模式的初步研究[D]. 武汉: 华中农业大学, 2009.
[37]	Debeaujon I, Nesi N, Perez P, et al.Proanthocyanidin-accumulating cells in Arabidopsis testa: regulation of differentiation and role in seed development[J]. Plant Cell, 2003, 15(11): 2514-2431.
[38]	Baudry A, Caboche M, Lepiniec L.TT8 controls its own expression in a feedback regulation involving TTG1 and homologous MYB and bHLH factors, allowing a strong and cell-specific accumulation of fiavonoids in Arabidopsis thaliana[J]. Plant J, 2006, 46(5): 768-779.
[39]	Quattrocchio F, Wing J F, Leppen HTC, et al.Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes[J]. Plant Cell, 1993, 5(11): 1497-1512.
[40]	de Vetten N, Quattrocchio F, Mol J, et al. The an11 locus controlling fiower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast, plants, and animals[J]. Genes Dev, 1997, 11(11): 1422-1434.
[41]	Spelt C, Quattrocchio F, Mol J, et al.ANTHOCYANIN1 of petunia controls pigment synthesis, vacuolar pH, and seed coat development by genetically distinct mech anisms[J]. Plant Cell, 2002, 14(9): 2121-2135.