欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2016, Vol. 42 ›› Issue (01): 1-10.doi: 10.3724/SP.J.1006.2016.00001

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

GmMYB042基因对类黄酮生物合成的调控作用

杜海1，冉凤1，马珊珊1，柯蕴倬1，孙丽萍1，李加纳1,*，唐益雄2,*   

  1. 1西南大学农学与生物科技学院，重庆400715；2中国农业科学院生物技术研究所，北京100081
  • 收稿日期:2015-06-08 修回日期:2015-09-06 出版日期:2016-01-12 网络出版日期:2015-10-08
  • 通讯作者: 李加纳, E-mail: ljn1950@swu.edu.cn, Tel: 13509496702; 唐益雄, E-mail: yixiongtang@sohu.com, Tel: 010-62121506.
  • 基金资助:

    本研究由国家自然科学基金项目(31471528)，国家博士后基金面上项目(2014M552297)和中央高校基本科研业务费专项资金(SWU113104, XDJK2014B035)资助。

Regulating Effects of GmMYB042 Gene on Flavonoid Biosynthesis

DU Hai1, RAN Feng1,MA Shan-Shan1,KE Yun-Zhuo1,SUN Li-Ping1,LI Jia-Na1,*,TANG Yi-Xiong2,*   

  1. 1 College of Agronomy and Biotechnology, Southwest University, Chongqing 400716, China; 2 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2015-06-08 Revised:2015-09-06 Published:2016-01-12 Published online:2015-10-08
  • Supported by:

    This research was supported by National Natural Science Foundation of China (31471528), Postdoctoral Science Foundation of China (2014M552297) and Fundamental Research Funds for the Central Universities of China (SWU113104, XDJK2014B035).

RichHTML

PDF

1576

被引次数

2

摘要：

MYB类转录因子是植物中最大的转录因子基因家族之一，广泛参与植物生长发育全过程，对植物次生代谢等具有重要的调控作用。本研究对大豆GmMYB042基因的表达特性和功能进行了系统的研究，针对该基因C端的保守氨基酸基序(PDLNLELTIS)和锌指结构进行了一系列的序列删除突变，并将各缺失突变体在烟草中进行了过表达，以验证目的基因及其特殊基序的功能。表达特性分析结果表明GmMYB042基因在大豆的根瘤、根、茎、叶、花、荚果皮和种子中均有表达，且在茎、种子和花中的表达量相对较高；GmMYB042基因在大豆中的表达受PEG、高盐、低温和UV-B辐射的诱导。过表达分析结果表明，GmMYB042基因的过表达使转基因烟草类黄酮代谢途径部分关键酶基因(如PAL、CHS和FLS)的表达量明显上升，转基因烟草总黄酮的含量明显高于对照；各缺失突变体的转基因烟草类黄酮代谢途径相应酶基因的表达量发生相应的变化，进一步证明目的基因对类黄酮生物合成的调控作用；各缺失突变体的转基因烟草的叶缘有明显的皱褶，说明目的基因可能还参与调控叶的形态建成。

关键词: MYB转录因子, 类黄酮, 缺失突变, 功能研究

Abstract:

MYB transcription factor is one of the largest transcription factor gene families in land plants, and is involved in a myriad of regulatory processes, such as secondary metabolism. In the present study, the expression profiles and function of GmMYB042 gene were systematically studied. In order to investigate the roles of the conserved amino acid motif PDLNLELTIS and a predicted zinc finger region at its C-terminal, a series of sequence deletions of these two regions were made by PCR method. Subsequently, the corresponding over-expression constructs of GmMYB042 gene and its mutants were made and transformed into tobacco NC89 with Agrobacterium LBA4404, respectively. Expression analyses revealed that GmMYB042 gene was expressed in nodule, root, stem, leaf, flower, pod and seed of soybean, and with a relative higher expression level in stem, flower and seed; its expression could be induced by PEG, high salt, low temperature and UV-B radiation stresses. Over-expression analyses showed that the expressions of some enzyme genes in flavonoid biosynthesis pathway (including PAL, CHS, CHI, and FLS) were obviously increased in GmMYB042 transgenic lines, resulting in an increased content of the flavonoid compounds. Accordingly, the transcription levels of the corresponding enzyme genes involved in flavonoid biosynthesis pathway were decreased in the transgenic lines of GmMYB042 mutants, further supporting the conclusion of regulating role of GmMYB042 gene in tobacco flavonoid biosynthesis pathway.

Key words: MYB transcription factor, Flavonoid, Deletion mutation, Functional research

[1] Klempnauer K H, Gonda T J, Bishop J M. Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb: the architecture of a transduced oncogene. Cell, 1982, 31: 453–463



[2] Du H, Zhang L, Liu L, Tang X F, Yang W J, Wu Y M, Huang Y B, Tang Y X. Biochemical and molecular characterization of plant MYB transcription factor family. Biochemistry (Mosc), 2009, 74: 1–11



[3] Du H, Wang Y B, Xie Y, Liang Z, Jiang S J, Zhang S S, Huang Y B, Tang Y X. Genome-wide identification and evolutionary and expression analyses of MYB-related genes in land plants. DNA Res, 20: 437–448



[4] Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L. MYB transcription factors in Arabidopsis. Trends Plant Sci, 2010, 15: 573–581



[5] Weston K. Myb proteins in life, death and differentiation. Curr Opin Genet Dev, 1998, 8: 76–81



[6] Paz-Ares J, Ghosal D, Wienand U, Peterson P A, Saedler H. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J, 1987, 6: 3553–3558



[7] Stracke R, Werber M, Weisshaar B. The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol, 2001, 4: 447–456



[8] Du H, Yang S S, Liang Z, Feng B R, Liu L, Huang Y B, Tang Y X. Genome-wide analysis of the MYB transcription factor superfamily in soybean. BMC Plant Biol, 2012, 12:106



[9] Wilkins O, Nahal H, Foong J, Provart N J, Campbell M M. Expansion and diversification of the Populus R2R3-MYB family of transcription factors. Plant Physiol, 2009, 149: 981–993



[10] Stracke R, Jahns O, Keck M, Tohge T, Niehaus K, Fernie A R, Weisshaar B. Analysis of PRODUCTION OF FLAVONOL GLYCOSIDES-dependent flavonol glycoside accumulation in Arabidopsis thaliana plants reveals MYB11-, MYB12- and MYB111-independent flavonol glycoside accumulation. New Phytol, 2010, 188: 985–1000



[11] Chen B J, Wang Y, Hu Y L, Wu Q, Lin Z P. Cloning and characterization of a drought-inducible MYB gene from Boea crassifolia. Plant Sci, 2005, 168: 493–500



[12] Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant cell, 2003, 15: 63–78



[13] Cominelli E, Galbiati M, Vavasseur A, Conti L, Sala T, Vuylsteke M, Leonhardt N, Dellaporta S L, Tonelli C. A guard-cell-specific MYB transcription factor regulates stomatal movements and plant drought tolerance. Curr Biol, 2005, 15: 1196–1200



[14] Seo P J, Xiang F, Qiao M, Park J Y, Lee Y N, Kim S G, Lee Y H, Park W J, Park C M. The MYB96 transcription factor mediates abscisic acid signaling during drought stress response in Arabidopsis. Plant Physiol, 2009, 151: 275–289



[15] Oh J E, Kwon Y, Kim J H, Noh H, Hong S W, Lee H. A dual role for MYB60 in stomatal regulation and root growth of Arabidopsis thaliana under drought stress. Plant Mol Biol, 2011, 77: 91–103



[16] Du H, Huang Y, Tang Y. Genetic and metabolic engineering of isoflavonoid biosynthesis. Appl Microbiol Biotechnol, 2010, 86:1293–1312



[17] Buer C S, Imin N, Djordjevic M A. Flavonoids: new roles for old molecules. J Integr Plant Biol, 2010, 52: 98–111



[18] Yang C S, Landau J M, Huang M T, Newmark H L. Inhibition of carcinogenesis by dietary polyphenolic compounds. Annu Rev Nutr, 2001, 21: 381–406



[19] Ross J A, Kasum C M.: Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu Rev Nutr, 2002, 22: 19–34



[20] Jin H, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C. Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. EMBO J, 2000, 19: 6150–6161



[21] Legay S, Sivadon P, Blervacq A S, Pavy N, Baghdady A, Tremblay L, Levasseur C, Ladouce N, Lapierre C, Séguin A, Hawkins S, Mackay J, Grima-Pettenati J. EgMYB1, an R2R3 MYB transcription factor from eucalyptus negatively regulates secondary cell wall formation in Arabidopsis and poplar. New Phytol, 2010, 188: 774–786



[23] Sonbol F M, Fornalé S, Capellades M, Encina A, Touriño S, Torres J L, Rovira P, Ruel K, Puigdomènech P, Rigau J, Caparrós-Ruiz D. The maize ZmMYB42 represses the phenylpropanoid pathway and affects the cell wall structure, composition and degradability in Arabidopsis thaliana. Plant Mol Biol, 2009, 70: 283–296



[24] Mehrtens F, Kranz H, Bednarek P, Weisshaar B. The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. Plant Physiol, 2005, 138: 1083–1096



[25] Stracke R, Ishihara H, Huep G, Barsch A, Mehrtens F, Niehaus K, Weisshaar B. Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. Plant J, 2007, 50: 660–677



[26] Czemmel S, Stracke R, Weisshaar B, Cordon N, Harris N N, Walker A R, Robinson S P, Bogs J. The grapevine R2R3-MYB transcription factor VvMYBF1 regulates flavonol synthesis in developing grape berries. Plant Physiol, 2009, 151: 1513–1530



[27] 杨文杰, 杜海, 方芳, 杨婉身, 吴燕民, 唐益雄. 大豆两个MYB 转录因子基因的克隆及表达分析. 中国农业科学, 2008, 41: 961–970



Yang W J, Du H, Fang F, Yang W S, Wu Y M, Tang Y X. Cloning and characterization of two new MYB transcription factor genes from soybean. Sci Agric Sin, 2008, 41: 961–970 (in Chinese with English abstract)



[28] 杜海, 刘蕾, 唐晓凤, 高杰, 杨文杰, 吴燕民, 黄玉碧, 唐益雄. 大豆MYB 转录因子GmMYBZ2 的鉴定、突变分析及原核. 农业生物技术学报, 2009, 17: 301–306



Du H, Liu L, Tang X F, Gao J, Yang W J, Wu Y M, Huang Y B, Tang Y X. Characterization, expression and transcriptional activation analysis of GmMYBZ2 in soybean. J Agric Biotechnol, 2009, 17: 301–306 (in Chinese with English abstract)



[29] Severin A J, Woody J L, Bolon Y T, Joseph B, Diers B W, Farmer A D, Muehlbauer G J, Nelson R T, Grant D, Specht J E, Graham M A, Cannon S B, May G D, Vance C P, Shoemaker R C. RNA-Seq Atlas of Glycine max: a guide to the soybean transcriptome. BMC Plant Biol, 2010, 10: 160



[30] Libault M, Farmer A, Joshi T, Takahashi K, Langley R J, Franklin L D, He J, Xu D, May G, Stacey G. An integrated transcriptome atlas of the crop model Glycine max, and its use in comparative analyses in plants. Plant J, 2010, 63: 86–99



[31] Lodovico T, Angel M, Adrian P, Mackay S, Francisco A, Culianez-Macia, Keith R, Cathie M. The AmMYB308 and AmMYB330 transcription factors from antirrhinum regulate phenylpropanoid and lignin biosynthesis in transgenic tobacco. Plant Cell, 1998, 10: 135–154



[32] Jin H, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C.Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. EMBO J, 2000, 19: 6150–6161



[33] Du H, Liang Z, Zhao S, Nan M G, Phan Tran L S, Lu K, Huang Y B, Li J N. The evolutionary history of R2R3-MYB proteins across 50 eukaryotes: new insights into subfamily classification and expansion. Sci Rep, 2015, 5: 11037
[1] 杨德卫, 王勋, 郑星星, 项信权, 崔海涛, 李生平, 唐定中. OsSAMS1在水稻稻瘟病抗性中的功能研究[J]. 作物学报, 2022, 48(5): 1119-1128.
[2] 黄伟, 高国应, 吴金锋, 刘丽莉, 张大为, 周定港, 成洪涛, 张凯旋, 周美亮, 李莓, 严明理. 芥菜型油菜BjA09.TT8和BjB08.TT8基因调节类黄酮的合成[J]. 作物学报, 2022, 48(5): 1169-1180.
[3] 邱红梅, 陈亮, 侯云龙, 王新风, 陈健, 马晓萍, 崔正果, 张玲, 胡金海, 王跃强, 邱丽娟. 大豆种子颜色遗传调控机制研究进展[J]. 作物学报, 2021, 47(12): 2299-2313.
[4] 高国应, 伍小方, 黄伟, 周定港, 张大为, 周美亮, 张凯旋, 严明理. 芥菜型油菜BjuB.KAN4基因调控类黄酮的途径[J]. 作物学报, 2020, 46(9): 1322-1331.
[5] 王艳花, 荐红举, 邱晓, 李加纳. 白菜型油菜粒色主效基因BrTT1的调控机制分析[J]. 作物学报, 2020, 46(11): 1678-1689.
[6] 黄启秀,曲延英,姚正培,李梦雨,陈全家*. 海岛棉枯萎病抗性与类黄酮代谢相关基因表达量的相关[J]. 作物学报, 2017, 43(12): 1791-1801.
[7] 单天雷,洪彦涛,杜丽璞,徐惠君,魏学宁,张增艳. 抗根腐病的TaMYB86过表达转基因小麦的创制与分子功能鉴定[J]. 作物学报, 2016, 42(10): 1429-1436.
[8] 高巍**,刘会利**,田新权,张慧,宋洁,杨勇,龙璐,宋纯鹏*. 海岛棉转录因子基因GbMYB60的克隆、表达及其抗逆性分析[J]. 作物学报, 2016, 42(09): 1342-1351.
[9] 张星星,王召军,杨玉双,王道文,郑文明,董振营. 利用HMW-GS全缺失突变体快速构建Glu-1位点近等渗入系[J]. 作物学报, 2016, 42(08): 1247-1252.
[10] 何海旺，潘华清，张铙丹，何龙飞*. 飞机草类黄酮3’-羟化酶基因(CoF3’H)的克隆及其在烟草中的表达[J]. 作物学报, 2015, 41(03): 479-486.
[11] 曲存民,卢坤,刘水燕,卜海东,付福友,王瑞,徐新福,李加纳. 黄黑籽甘蓝型油菜类黄酮途径基因SNP位点检测分析[J]. 作物学报, 2014, 40(11): 1914-1924.
[12] 王诺菡,于霁雯,吴嫚,马启峰,李兴丽,裴文锋,李海晶,黄双领,张金发,喻树迅. 棉花GhMYB0基因的克隆、表达分析及功能鉴定[J]. 作物学报, 2014, 40(09): 1540-1548.
[13] 夏家平，郭会君，谢永盾，赵林姝，古佳玉，赵世荣，李军辉，刘录祥. 小麦叶绿素缺失突变体Mt135的叶绿体基因差异表达分析[J]. 作物学报, 2012, 38(11): 2122-2130.
[14] 宋素洁，古佳玉，郭会君，赵林姝，赵世荣，李军辉，赵宝存，刘录祥. 小麦叶绿素缺失突变体Mt6172及其野生型叶片蛋白质组学双向差异凝胶电泳分析[J]. 作物学报, 2012, 38(09): 1592-1606.
[15] 吴磊，王丹，苏文悦，郭长虹，束永俊. 利用比较基因组学开发山羊草属InDel分子标记[J]. 作物学报, 2012, 38(07): 1334-1338.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2